Progress on Theme 1: Animal Models to Advance the Study of Human Disease
Programs and Activities Highlights
- Invertebrate Models Fact Sheet
ORIP revised its fact sheet on invertebrate models. ORIP reviewed the existing section for accuracy and added a link to “MAGIC tools,” a genetic tool for genome-wide mosaic analysis of Drosophila resources. This fact sheet is one of several fact sheets that serve as valuable resources for potential investigators to learn about ORIP resources and programs. - Nonhuman Primate Resources Fact Sheet
ORIP revised its fact sheet on Nonhuman Primate Resources. ORIP reviewed the fact sheet for accuracy and added information on two resources: the Biospecimens Query System of the National Primate Research Centers Consortium and the Primate Pathology Image Database. This is one of several fact sheets that serve as valuable resources for potential investigators to learn about ORIP resources and programs. - Swine Models Fact Sheet
ORIP created a new fact sheet on Swine Models. Swine have become valuable models for cardiovascular diseases, diabetes, heart and lung transplantation, and xenotransplantation. This fact sheet lists valuable ORIP-supported resources and initiatives in a concise and comprehensive manner. This is one of several fact sheets that serve as valuable resources for potential investigators to learn about ORIP resources and programs. - National Primate Research Centers Directors Biannual Meeting
The National Primate Research Centers (NPRCs) Directors Meeting was held in spring 2024. During this meeting, the Directors and other NPRC staff provided information and updates on their centers. The NPRC Program complements and enables the missions of NIH institutes and centers by providing the animals, facilities, expertise, and resources for nonhuman primate research in specific disease areas. - 2024 U42 Specific-Pathogen-Free Macaque Colonies Steering Committee Meeting
ORIP scheduled and organized the annual meeting of the U42 Specific-Pathogen-Free (SPF) Macaque Steering Committee held May 8, 2024. Attendees included U42 SPF macaque colony principal investigators, other key people associated with the U42 SPF colonies, and OAR and NIH staff. Information presented during the meeting will help ORIP make future recommendations and budget justifications for new initiatives and programs. - Rodent Resource Fact Sheet
ORIP revised its fact sheet on rodent resources. ORIP reviewed the fact sheet for accuracy and added artificial intelligence–based mouse genetic discovery as a resource. This fact sheet is one of several fact sheets that serve as valuable resources for potential investigators to learn about ORIP resources and programs. - Site Visit to Emory National Primate Research Center
The ORIP and DPCPSI Directors participated in a site visit to the Emory National Primate Research Center (ENPRC) in March 2024. They viewed the facilities and procedures at ENPRC to improve communications, understanding, and trust between partners. They also reinforced the team approach between ENPRC and NIH to addressing challenges. - Notice of Extension of the Expiration Date for PAR-21-167: Development of Animal Models and Related Biological Materials for Research (R21 Clinical Trial Not Allowed)
The purpose of this notice is to extend the expiration date for PAR-21-167, Development of Animal Models and Related Biological Materials for Research (R21 Clinical Trial Not Allowed). With the addition/extension by two application due dates, PAR-21-167 now expires on January 8, 2025. NIH standard due dates are applied to this notice. - Discussion Regarding Conventional Nonhuman Primates
ORIP convened a meeting among the California National Primate Research Center (CNPRC), Biomedical Advanced Research and Development Authority, Centers for Disease Control and Prevention, U.S. Department of Defense, U.S. Food and Drug Administration, and NIH collaborators in February 2024 to discuss CNPRC's availability of conventional NHPs for biomedical research. - Limited Competition: Specific-Pathogen-Free Macaque Colonies to Support HIV/AIDS Research (U42 Clinical Trial Not Allowed); PAR-24-129
The purpose of this notice of funding opportunity is to provide continuing support for specific-pathogen-free (SPF) macaque colonies previously funded under the auspices of PAR-21-089 and PAR-18-669 that support HIV research. Breeding colonies are essential to sustain appropriate SPF macaques for research related to HIV. Pedigree SPF macaques are free of certain viruses, which may confound the results of HIV-related investigations or present a risk to the personnel who care for the animals. The SPF macaques are genetically characterized for major histocompatibility complex class I types, which have large effects on macaque immune responses to simian immunodeficiency virus. - Sex as a Biological Variable Policy for Studies Using Nonhuman Primates in Neuroscience Research
The NIH Office of Research on Women’s Health developed the NIH Policy on Sex as a Biological Variable (SABV), which builds on the importance of sex in biology, health, and disease, as well as the need for research to consider sex to be relevant for all people. In January 2024, NIH convened a virtual workshop to provide the neuroscience research community an opportunity to discuss challenges and potential solutions for applying the SABV policy fairly and consistently to nonhuman primate (NHP) studies. An ORIP staff member presented at the workshop as an invited panelist to provide ORIP's perspective on the NHP shortage, best utilization of existing resources, and issues regarding NHP use in neuroscience research. - BioGRID: Resource Overview
ORIP published a video overview of the Biological General Repository for Interaction Datasets (BioGRID) program in January 2024. BioGRID is an open-access public database that uses structured curation to capture protein, genetic, and chemical interaction data from model organisms and humans. - Limited Competition: Mutant Mouse Resource and Research Centers (U42 Clinical Trial Not Allowed) PAR-24-105
This notice of funding opportunity invites applications for the continued support and advancement of the Mutant Mouse Resource and Research Centers (MMRRCs). The MMRRC consortium is expected to facilitate research by identifying, acquiring, evaluating, characterizing, cryopreserving, and distributing mutant mouse strains to qualified biomedical investigators. A regional network of four MMRRCs and an Informatics, Coordination and Service Center collectively serve the needs of the biomedical research community for transgenic, knockout, and other genetically engineered mutant mice and related biomaterials. MMRRC strains are held to the highest standards to optimize reproducibility of studies and ensure scientific rigor and transparency; all submitted strains are thoroughly reviewed and documented and include additional quality control measures. The Program Director/Principal Investigator of each MMRRC is required to develop a small high-risk, high-return, research pilot project that complements the goals and needs of the MMRRC consortium. - Conjugation of HIV-1 Envelope to Hepatitis B Surface Antigen Alters Vaccine Responses in Rhesus Macaques
Researchers are interested in developing an HIV-1 vaccine that improves upon the regimen used in the RV144 clinical trial. The authors tested the hypothesis that a conjugate vaccine based on the learned response to immunization with hepatitis B virus could be utilized to expand T-cell help and improve antibody production against HIV-1. Using juvenile rhesus macaques of both sexes, they evaluated the immunogenicity of their conjugate regimen. Their findings suggest that conjugate vaccination can engage both HIV-1 Env–specific and hepatitis B surface antigen–specific T-cell help and modify antibody responses at early time points. This work may help inform future efforts to improve the durability and efficacy of next-generation HIV vaccines. - The Monarch Initiative in 2024: An Analytic Platform Integrating Phenotypes, Genes and Diseases Across Species
The Monarch Initiative aims to bridge the gap between the genetic variations, environmental determinants, and phenotypic outcomes critical for translational research. The Monarch app provides researchers access to curated data sets with information on genes, phenotypes, and diseases across species and advanced analysis tools for such diverse applications as variant prioritization, deep phenotyping, and patient profile matching. Researchers describe upgrades to the app, including scalable cloud-based infrastructure, simplified data ingestion and knowledge graph integration systems, enhanced data mapping and integration standards, and a new user interface with novel search and graph navigation features. A customized plugin for OpenAI’s ChatGPT allows the use of large language models to interrogate knowledge in the Monarch graph and increase the reliability of the responses of Monarch’s analytic tools. These upgrades will enhance clinical diagnosis and the understanding of disease mechanisms. - Conduction-Dominated Cryomesh for Organism Vitrification
Vitrification-based cryopreservation via cryomesh is a promising approach for maintaining biodiversity, health care, and sustainable food production via long-term preservation of biological systems. The researchers conducted a series of experiments aimed at optimizing the cooling and rewarming rates of cryomesh to increase the viability of various cryopreserved biosystems. They found that vitrification was significantly improved by increasing thermal conductivity, reducing mesh wire diameter and pore size, and minimizing the nitrogen vapor barrier of the conduction-dominated cryomesh. Cooling rates increased twofold to tenfold in a variety of biosystems. The conduction-dominated cryomesh improved the cryopreservation outcomes of coral larvae, Drosophila embryos, and zebrafish embryos by vitrification. These findings suggest that the conduction-dominated cryomesh can improve vitrification in such biosystems for biorepositories, agriculture and aquaculture, and research. - Investigation of Monoclonal Antibody CSX-1004 for Fentanyl Overdose
The opioid crisis in the United States is primarily driven by the highly potent synthetic opioid fentanyl and has led to more than 70,000 overdose deaths annually; thus, new therapies for fentanyl overdose are urgently needed. The authors present the first clinic-ready, fully human monoclonal antibody CSX-1004 with picomolar affinity for fentanyl and related analogs. In mice of both sexes, CSX-1004 reverses fentanyl antinociception and the intractable respiratory depression caused by the ultrapotent opioid carfentanil. Using a highly translational nonhuman primate model, squirrel monkeys of both sexes, for respiratory depression, they demonstrate CSX-1004-mediated protection from repeated fentanyl challenges for 3–4 weeks. These data establish the feasibility of CSX-1004 as a promising candidate medication for preventing and reversing fentanyl-induced overdose. - A Combined Adjuvant Approach Primes Robust Germinal Center Responses and Humoral Immunity in Non-Human Primates
Protein antigens require adjuvants for high immunogenicity, and delivery kinetics are a critical component of rational HIV vaccine design. Investigators employed a combined adjuvant approach (i.e., short phosphoserine peptide linkers that promote tight binding to aluminum hydroxide, plus saponin/MPLA nanoparticles) with slow antigen delivery and potent immune-stimulating complexes in rhesus macaques of both sexes. They reported that pSer-modified antigen shifts immunodominance to allow subdominant epitope-targeting of rare B cells. These findings indicate that a combined adjuvant approach can augment humoral immunity by modulating immunodominance, and this work can be applied for the development of clinical therapeutics. - Oncology Models Forum Annual Meeting
An ORIP staff member presented ORIP’s rodent resources at the Oncology Models Forum Annual Meeting in December 2023. Topics of discussion at the meeting included discussed obstacles to answering translational questions in models, as well as potential approaches to overcome those obstacles. - Knockout Mouse Phenotyping Program/International Mouse Phenotyping Consortium: Annual Fall 2023 Meeting
An ORIP program staff member attended the Knockout Mouse Phenotyping Program (KOMP2)/International Mouse Phenotyping Consortium (IMPC) Annual Fall Meeting, which was held in Houston, Texas, in November 2023. KOMP2 collaborates with IMPC to knockout and characterize all protein-coding genes in the mouse genome. ORIP’s participation in this meeting was required for monitoring the overall project progress and facilitating the exchange of scientific knowledge with international collaborators. - Fall 2023 Human Tissue and Organ Research Resource Steering Committee Meeting
ORIP program staff attended the Fall 2023 Meeting of the Human Tissue and Organ Research Resource (HTORR) Steering Committee held on November 29, 2023. HTORR, supported by grant U42OD011158, provides high-quality human biospecimens to investigators to facilitate scientific advances in biomedical research across multiple disciplines. HTORR programmatic updates were presented to the steering committee. - 21st Annual Meeting of the Mutant Mouse Resource and Research Center Consortium
An ORIP program staff member attended the 21st Annual Meeting of the Mutant Mouse Resource and Research Center (MMRRC) Consortium in October 2023. The staff member presented an NIH update, participated in the discussions, advised on policies, evaluated the MMRRC program progress, and interacted with external advisory committee members. - Complement Animal Research in Experimentation (Complement-ARIE) Program
An ORIP program staff member served as a co-facilitator at the public listening session for the Complement ARIE strategic planning meeting on October 2, 2023. Complement ARIE is focused on development, standardization, validation, and use of methods and approaches that will more accurately model human biology, known as new approach methodologies (NAMs). The listening session included key representatives from multiple sectors to gain insight into current opportunities and challenges regarding NAMs development. - ORIP Strategic Planning Retreat
ORIP staff members participated in a strategic planning retreat on August 25, 2023. Topics of discussion included scientific stewardship and progress on ORIP’s current strategic plan, awareness of ORIP resources and programs, animal models to advance the study of human disease, innovative instruments and equipment to accelerate research discoveries, specialized research training in animal models and related resources, and the strategic plan document layout. Key themes of the discussion included outreach and communication, thoughtful evaluation of ORIP programs, and data collection and curation. - Knockout Mouse Phenotyping Program (KOMP2)
KOMP2 collaborates with the International Mouse Phenotyping Consortium (IMPC) to knockout and characterize all protein-coding genes in the mouse genome. ORIP is providing support to KOMP2 through operational decision-making and is helping to oversee interactions with IMPC partners. An ORIP staff member recently participated in a KOMP2 meeting, which was held to monitor the project’s overall progress, foster exchange of scientific knowledge with international collaborators, facilitate discussions with production and phenotyping center representatives, conduct an in-depth review of data, and assess the progress and the impact of the projects. ORIP also conducted a site visit to the Mary Lyon Centre, the United Kingdom’s national facility for mouse genetics. - International Mouse Phenotyping Consortium Annual Conference
The annual conference of the International Mouse Phenotyping Consortium and the Knockout Mouse Phenotyping Program was held in July 2023. ORIP staff members participated in the meeting organization, invited speakers, participated in discussions, monitored program progress and impact, and guided the grant awardees. - Association of Biomolecular Resource Facilities 2023 Annual Meeting
An ORIP program staff member delivered a presentation on the NIH S10 Shared Instrumentation Grant Program at the Association of Biomolecular Resource Facilities 2023 Annual Meeting on May 8, 2023. The presentation was given to a diverse audience and showcased recent accomplishments of the S10 programs in supporting operations of core facilities across research institutions nationwide. - Site Visit to the Aquatic Germplasm and Genetic Resources Center
ORIP conducted a site visit to the Aquatic Germplasm and Genetic Resources Center at the Louisiana State University Agricultural Center in April 2023. The purpose of this site visit was to oversee the impact and effectiveness of ORIP’s investments in the Center during the past 3 years. - P40 Pre-Application Webinar
ORIP held a pre-application webinar on February 28, 2023, to provide information on the Notice of Special Interest: Revision Applications to Add a Curation and Informatics Component to Existing Animal and Biological Material Resource Centers (NOT‑OD-23-068). Following the webinar, the meeting slides and frequently asked questions were made available on the ORIP website. - Notice of Funding Opportunity (NOFO): PAR-22-252 (Early-Stage Investigator HIV/AIDS Research Using Nonhuman Primate (NHP) Models (R21, Clinical Trial Not Allowed))
ORIP and the National Institute of Mental Health published an R21 NOFO to support preclinical HIV/AIDS research performed by early‑stage investigators using NHP models. The purpose of this NOFO is to provide early-stage investigators adequate funds and independence to initiate preclinical and translational HIV/AIDS research using NHPs. As a result of this support, these investigators should become competitive for major research awards (e.g., R01) and for transition to independent positions, which will help advance HIV/AIDS researchers trained to use NHPs to fill this national need. - Mutant Mouse Resource & Research Centers (MMRRC) Annual Meeting
The annual MMRRC meeting took place at The Jackson Laboratory facility in Bar Harbor, Maine, in November 2022. The four Centers (University of California, Davis; University of Missouri; The University of North Carolina at Chapel Hill; and The Jackson Laboratory) and the Informatics, Coordination and Service Center were represented, as well as the Advisory Board. ORIP participated in organizing the meeting, presented an NIH update, and participated in the discussions. - Funding Opportunity Announcement (FOA): PAR-23-040 (Development of Resources and Technologies for Enhancing Rigor, Reproducibility, and Translatability of Animal Models in Biomedical Research (R01))
ORIP published an R01 FOA encouraging research project grant applications aimed at developing or improving technologies, tools, and resources for validating animal models and improving rigor, reproducibility, and translatability of animal research, which will be broadly applicable to biomedical research involving animal models. Animal models of interest for this FOA include, but are not limited to, invertebrate and vertebrate organisms ranging from Caenorhabditis elegans and Drosophila to zebrafish, mice, rats, pigs, and nonhuman primates. - Funding Opportunity Announcement (FOA): PAR-23-039 (Development of Resources and Technologies for Enhancing Rigor, Reproducibility, and Translatability of Animal Models in Biomedical Research (R24 Clinical Trials Not Allowed))
ORIP published an R24 FOA encouraging resource-related research grant applications that address key animal resource- and technology-related gaps, which include the need for systematic phenotyping of animal models, improved genetic technologies, high-throughput screening, and integration of multiple research organisms. Applications submitted under this FOA should aim to enhance the rigor, reproducibility, and translatability of animal research through the development of resources and technologies that have significant impact across a broad range of research areas using animal models. - Fall 2022 Human Tissue and Organ Research Resource (HTORR) Steering Committee Meeting
ORIP organized and conducted the HTORR Steering Committee Meeting in November 2022. ORIP participated in discussions with HTORR’s principal investigator and other staff from the National Disease Research Interchange, a not-for-profit organization; NIH members of the steering committee; and other NIH guests. In attendance were NIH program staff from the National Institute of Allergy and Infectious Diseases; National Heart, Lung, and Blood Institute; National Institute of Diabetes and Digestive and Kidney Diseases; National Eye Institute; National Institute of Arthritis and Musculoskeletal and Skin Diseases; National Institute on Aging; and National Institute of Mental Health. - National Xenopus Resource Site Visit
ORIP conducted a site visit to the National Xenopus Resource on October 17, 2022, to evaluate the impact of four Alteration and Renovation Administrative Supplements awarded during the past 3 years through the Resource’s P40 grant and a related R24 grant. - Funding Opportunity Announcement: Development of Animal Models and Related Biological Materials for Down Syndrome Research (R24 Clinical Trials Not Allowed)
The INvestigation of Co-occurring conditions across the Lifespan to Understand Down syndromE (INCLUDE) Project encourages grant applications aimed at developing, characterizing, or improving animal models and related biological materials for Down syndrome. ORIP is partnering with seven other Institutes, Centers, and Offices in this funding opportunity announcement. - California National Primate Research Center (CNPRC) Site Visit
ORIP program staff participated in a site visit to the CNPRC on August 10, 2022. The visit and peer review of the CNPRC were led by a Scientific Review Officer at the Center for Scientific Review. - ORIP Concept Clearance: Reissuance—Resource-Related Research Projects for Development of Animal Models and Related Materials
ORIP presented this concept clearance for reissue at the September 2022 NIH Council of Councils meeting. The initiative will encourage grant applications for developing, characterizing, or improving animal models of human diseases; developing or improving technologies and methods that aim to enhance rigor and reproducibility of research with animal models; improving access to information about or generated from the use of animal models of human disease; or improving diagnosis and control of diseases of laboratory animals. - ORIP Concept Clearance: Somatic Cell Genome Editing (SCGE) Testing Center
ORIP presented this concept clearance at the September 2022 NIH Council of Councils meeting. Based on recommendations from previous workshops and ORIP’s oversight of animal testing centers and reporter systems projects for the NIH-wide SCGE Program, ORIP proposes to initiate a new program titled “Testing Centers for Development of Somatic Cell Genome Editing in Model Organisms.” To align with ORIP’s NIH-wide mission, proposed projects will (1) cover a wide variety of disease conditions relevant to the interests of multiple NIH Institutes and Centers, (2) develop resources and testing services in animal models for the growing community developing SCGE therapeutics, (3) conduct testing in reporter animals and animal disease models, and (4) assist in developing new technologies and preclinical testing to generate high-quality, reproducible information required for clinical studies. - Funding Opportunity Announcements (FOAs): RFA-OD-22-013 (Resource-Related Research Projects for Development of Animal Models and Related Materials (R24 Clinical Trials Not Allowed)) and PAR-22-204 (Development of Animal Models and Related Materials for HIV/AIDS Research (R24 Clinical Trials Not Allowed))
ORIP published two R24 FOAs in September 2022 encouraging grant applications aimed at developing, characterizing, or improving animal models for human diseases (including HIV/AIDS); improving access to information about or generated from the use of animal models of human disease; or improving diagnosis and control of diseases of laboratory animals. - Omnibus Solicitation of the NIH, Centers for Disease Control and Prevention, and U.S. Food and Drug Administration for Small Business Innovation Research (SBIR) Grant Applications (Parent SBIR [R43/R44] Clinical Trial Not Allowed), PA-22-176; and Small Business Technology Transfer (STTR) Grant Applications (Parent STTR [R41/R42] Clinical Trial Not Allowed), PA-22-178
ORIP is participating in the SBIR and STTR Omnibus Solicitations/Funding Opportunity Announcements to support small business research aimed at developing biomedical methods and technologies that relate to improvements in animal models for human disease, as well as the care, use, and management of laboratory animals - Human Tissue and Organ Research Resource (HTORR) Steering Committee Meeting
The biannual meeting of the HTORR Steering Committee was held in spring 2022. HTORR, an ORIP-supported resource, provides high-quality human biospecimens to investigators to facilitate scientific advances in biomedical research across multiple disciplines. The format of the Steering Committee meeting was enhanced to include reports of recent scientific advancements—in areas of interest to multiple NIH Institutes and Centers (ICs)—that were made possible by HTORR-distributed biospecimens. This enhancement, which will continue for future meetings, was well received by the IC partners that support HTORR and increased their awareness of the impacts of co-funding support. - ORIP Concept Clearance: Development of Resources and Technologies for Enhancing Rigor, Reproducibility, and Translatability of Animal Models in Biomedical Research
ORIP presented this concept clearance at the May 2022 NIH Council of Councils meeting. The initiative will support research and resource-related research projects aimed at developing broadly applicable technologies, tools, and resources for validating animal models and enhancing rigor, reproducibility, and translatability of animal research. - ORIP Concept Clearance: Reissuance—Animal Models and Related Biological Materials for Research (R21)
ORIP presented this concept clearance for reissue at the May 2022 NIH Council of Councils meeting. The initiative encourages innovative research to develop, characterize, and improve animal models, biological materials, and novel technologies to better understand human health and disease. The initiative also seeks projects aimed at improving diagnosis and control of diseases that interfere with animal use for biomedical research. - National Primate Research Centers (NPRC) Directors Meeting
The NPRC Directors meeting, held in the spring of 2022, included updates from ORIP and the Division of Program Coordination, Planning, and Strategic Initiatives; updates from the NPRC Directors; and discussions of key activities, gaps, and opportunities related to the NPRC Program. - INCLUDE (INvestigation of Co-occurring conditions across the Lifespan to Understand Down syndromE) Project
ORIP is participating in INCLUDE, the NIH-wide initiative on Down syndrome research. In collaboration with other NIH Institutes, Offices, and Centers, ORIP signed on to the reissue of RFA-OD-20-005, titled, “Transformative Research Award for the INCLUDE Project (R01 Clinical Trial Not Allowed).” - Nonhuman Primate Developmental Genotype-Tissue Expression (NHP dGTEx) Research Center
ORIP is co-funding a National Human Genome Research Institute–managed dGTEx program, HG21-026, titled, “NHP dGTEx Project (U24 Clinical Trials Not Allowed).” The purpose of this funding opportunity is to solicit applications for the NHP dGTEx Research Center. The Center will perform four major functions: (1) create a tissue resource of multiple reference tissues across developmental stages in NHPs, (2) perform transcriptome sequencing and other genomic analysis of single-cell and bulk tissues, (3) analyze gene expression patterns in NHPs and compare them to human gene expression patterns, and (4) make the tissue samples and data available and usable for the research community. - Grassroots Efforts to End Structural Racism Throughout the U.S. National Institutes of Health
An open letter by NIH staff to the Director of the NIH helped to catalyze the NIH UNITE Initiative for Ending Structural Racism. ORIP staff participated in these grassroots efforts, as well as in the UNITE Initiative, to lead change within the NIH and the biomedical research ecosystem. - Severe Combined Immunodeficient Pig Models
ORIP supports development of immunodeficient swine research models and resources to enable biomedical research on a wide range of human diseases. Two ORIP staff members served as panelists at the January 2022 virtual meeting, “Severe Combined Immunodeficient (SCID) Pig Models: Industry and Academia’s Needs.” The panel discussed the need for novel SCID pig models in various fields, such as stem cell therapy, regenerative medicine, cancer therapeutics, and organ transplantation. In addition to funding the National Swine Resource and Research Center, ORIP is supporting two projects focusing on improvement of SCID pigs with natural mutation or creating new mutants using state-of-the-art technologies. - ORIP 2021–2022 Resource and Research Centers
ORIP supports 97 resource and research centers that develop animal models of human biology and disease. These centers develop, characterize, house, cryopreserve, and distribute both wild-type reference strains of animals, as well as mutant and genetically modified strains. - ORIP’s Rapid Response to the COVID-19 Pandemic
ORIP issued administrative supplements to many of its research resource centers in 2021 to support the development of new animal models and other resources specifically designed to facilitate research on SARS-CoV-2 or to support research projects related to COVID-19. - ORIP Workshop: Validation of Animal Models and Tools for Biomedical Research
ORIP—in collaboration with the National Heart, Lung, and Blood Institute; National Institute of Diabetes and Digestive and Kidney Diseases; National Institute of General Medical Sciences; National Institute of Neurological Disorders and Stroke; and National Institute on Aging—held a series of virtual workshops between November 2020 and January 2021 to discuss the status and needs regarding the validation and rigor/reproducibility of animal models used in biomedical research. - ORIP Encourages the Development of Resources and Technologies for Enhancing Rigor, Reproducibility, and Translatability of Animal Models in Biomedical Research
ORIP has issued a Notice of Special Interest (NOT-OD-22-039) for 2-year projects to support exploratory/developmental and highly innovative projects aimed at developing broadly applicable technologies, tools, and resources for validating animal models and enhancing the rigor, reproducibility, and translatability of animal research.
ORIP-Supported Research Highlights
- Systematic Multi-trait AAV Capsid Engineering for Efficient Gene Delivery
Engineering novel functions into proteins while retaining desired traits is a key challenge for developers of viral vectors, antibodies, and inhibitors of medical and industrial value. In this study, investigators developed Fit4Function, a generalizable machine learning (ML) approach for systematically engineering multi-trait adeno-associated virus (AAV) capsids. Fit4Function was used to generate reproducible screening data from a capsid library that samples the entire manufacturable sequence space. The Fit4Function data were used to train accurate sequence-to-function models, which were combined to develop a library of capsid candidates. Compared to AAV9, top candidates from the Fit4Function capsid library exhibited comparable production yields; more efficient murine liver transduction; up to 1,000-fold greater human hepatocyte transduction; and increased enrichment in a screen for liver transduction in macaques. The Fit4Function strategy enables prediction of peptide-modified AAV capsid traits across species and is a critical step toward assembling an ML atlas that predicts AAV capsid performance across dozens of traits. - The Mutant Mouse Resource and Research Center (MMRRC) Consortium: The U.S.-Based Public Mouse Repository System
The MMRRC has been the nation’s preeminent public repository and distribution archive of mutant mouse models for 25 years. The Consortium, with support from NIH, facilitates biomedical research by identifying, acquiring, evaluating, characterizing, preserving, and distributing a variety of mutant mouse strains to investigators around the world. Since its inception, the MMRRC has fulfilled more than 20,000 orders from 13,651 scientists at 8,441 institutions worldwide. Today, the MMRRC maintains an archive of mice, cryopreserved embryos and sperm, embryonic stem cell lines, and murine monoclonal antibodies for nearly 65,000 alleles. The Consortium also provides scientific consultation, technical assistance, genetic assays, microbiome analysis, analytical phenotyping, pathology, husbandry, breeding and colony management, and more. - Disruption of Myelin Structure and Oligodendrocyte Maturation in a Macaque Model of Congenital Zika Infection
Using a pigtail macaque model for Zika virus infection, researchers show that in utero exposure of a fetus to Zika virus due to maternal infection results in significantly decreased myelin formation around neurons. Myelin is a protective sheath that forms around neurons and is required for brain processing speed. Damaged myelin resulting from Zika infection in utero is likely a contributing factor to the severe deficits in brain development and microcephaly that result from maternal Zika virus infection. - AAV5 Delivery of CRISPR/Cas9 Mediates Genome Editing in the Lungs of Young Rhesus Monkeys
Genome editing in somatic cells and tissues has the potential to provide long-term expression of therapeutic proteins to treat a variety of genetic disorders of the lung. However, delivering genome-editing machinery to disease-relevant cell types in the lungs of primates has remained a challenge. Investigators of this article are participating in the NIH Somatic Cell Genome Editing Consortium. Herein, they demonstrate that intratracheal administration of a dual adeno-associated virus type 5 vector encoding CRISPR/Cas9 can mediate genome editing in rhesus (male and female) airways. Up to 8% editing was observed in lung lobes, including a housekeeping gene, GAPDH, and a disease-related gene, angiotensin-converting enzyme 2. Using single-nucleus RNA-sequencing, investigators systematically characterized cell types transduced by the vector. - Engineered IgM and IgG Cleaving Enzymes for Mitigating Antibody Neutralization and Complement Activation in AAV Gene Transfer
Recombinant adeno-associated viral (AAV) vectors have emerged as the leading platform for therapeutic gene transfer, but systemic dosing of AAV vectors poses potential risk of adverse side effects, including complement activation triggered by anti-capsid immunity. In this report, investigators discover an IgM cleaving enzyme (IceM) that degrades human IgM, a key trigger in the anti-AAV immune cascade. They engineered a fusion enzyme (IceMG) with dual proteolytic activity against human IgM and IgG. Antisera from female mice and rhesus macaques of both sexes treated with IceMG show decreased ability to neutralize AAV and activate complement. These studies have implications for improving the safety of AAV gene therapies, as well as broader potential applications, including organ transplantation and autoimmune diseases. - Single-Component Multilayered Self-Assembling Protein Nanoparticles Presenting Glycan-Trimmed Uncleaved Prefusion Optimized Envelope Trimers as HIV-1 Vaccine Candidates
Researchers are interested in engineering protein nanoparticles to mimic virus-like particles for an HIV-1 vaccine. In this study, researchers explored a strategy that combines HIV envelope glycoprotein (Env) stabilization, nanoparticle display, and glycan trimming. They designed a panel of constructs for biochemical, biophysical, and structural characterization. Using female mice, female rabbits, and rhesus macaques of both sexes, they demonstrated that glycan trimming increases the frequency of vaccine responders and steers antibody responses away from immunodominant glycan holes and glycan patches. This work offers a potential strategy for overcoming the challenges posed by the Env glycan shield in vaccine development. - Surgical Protocol for Partial Heart Transplantation in Growing Piglets
Researchers are interested in using partial heart transplantation (i.e., only the part of the heart containing the necessary heart valve is transplanted) to deliver growing heart valve implants. This novel technique allows partial heart transplants to grow, similar to the valves in heart transplants. More work is needed, however, to understand the underlying biological mechanisms of this approach and achieve progress in clinical care. In the current study, the authors present a surgical protocol for partial heart transplantation in growing piglets (sex not specified). This model will enable other researchers to seek fundamental knowledge about the nature of partial heart transplants. - CD8+ T Cell Targeting of Tumor Antigens Presented by HLA-E
Researchers have hypothesized that human leukocyte antigen-E (HLA-E)–positive cancer cells could be targeted by HLA-E–restricted CD8+ T cells. In this study, the authors assessed whether major histocompatibility complex E (MHC-E) expression by cancer cells can be targeted for MHC-E–restricted T-cell control. Using male rhesus macaques, they found that a cytomegalovirus can be used as a vector to generate specific immune cells that can target cancer cells. The authors conclude that targeting HLA-E with restricted specific CD8+ T cells could offer a new approach for immunotherapy of prostate cancer. Overall, this study supports the concept of a cancer vaccine. - Potent Antibody-Dependent Cellular Cytotoxicity of a V2-Specific Antibody Is Not Sufficient for Protection of Macaques Against SIV Challenge
Antibody-dependent cellular cytotoxicity (ADCC) has been correlated with decreased risk of HIV acquisition. Researchers tested the ability of PGT145, an antibody that neutralizes genetically diverse HIV-1 isolates, to protect male and female rhesus macaques against simian immunodeficiency virus (SIV) via ADCC activity. They found that a single amino acid substitution in the V2 core epitope of the SIV envelope increases PGT145 binding and confers sensitivity to neutralization. Peak and chronic phase viral loads were lower, and time to peak viremia was delayed. They concluded that ADCC is insufficient for protection by this antibody, but increasing the affinity of antibody binding could confer partial protection. - Consistent Survival in Consecutive Cases of Life-Supporting Porcine Kidney Xenotransplantation Using 10GE Source Pigs
Xenotransplantation offers potential for addressing organ donor shortages, and the U.S. Food and Drug Administration recently issued guidance on a regulatory path forward. Researchers have performed studies in this area, but concerns have been expressed about safe clinical translation of their results (e.g., survival, preclinical procurement, immunosuppression, clinical standards of care). In this study, the authors report consistent survival in consecutive cases of kidney xenotransplantation from pigs (male and female) to baboons (male and female). The authors propose that their findings serve as proof of concept for prevention of xenograft rejection in recipients of genetically modified porcine kidneys. This work offers insights for immunosuppression regimens for first-in-human clinical trials. - Identifying Potential Dietary Treatments for Inherited Metabolic Disorders Using Drosophila Nutrigenomics
Inherited metabolic disorders are known to cause severe neurological impairment and child mortality and can sometimes respond to dietary treatment; however, a suitable paradigm for testing diets is lacking for developing effective dietary treatment. In this study, researchers found that 26 of 35 Drosophila amino acid disorder models screened for disease–diet interactions displayed diet-altered development and/or survival. Among these male and female Drosophila models, researchers showed that dietary cysteine depletion normalizes metabolic profile and rescues development, neurophysiology, behavior, and life span in a model for isolated sulfite oxidase deficiency. These findings demonstrate the value of using Drosophila in studying diet-sensitive metabolic disorders and developing potential dietary therapies. - Cdk8/CDK19 Promotes Mitochondrial Fission Through Drp1 Phosphorylation and Can Phenotypically Suppress Pink1 Deficiency in Drosophila
Pink1 is a mitochondrial kinase implicated in Parkinson’s disease and is conserved among humans, rodents, and flies. In this study, researchers found that Cdk8 in male Drosophila (i.e., the orthologue of vertebrate CDK8 and CDK19) promotes the phosphorylation of Drp1 (i.e., a protein required for mitochondrial fission) at the same residue as Pink1. Cdk8 is expressed in both the cytoplasm and nucleus, and neuronal loss of Cdk8 reduces fly life span and causes bang sensitivity and elongated mitochondria in both muscles and neurons. Overexpression of Cdk8 suppresses elevated levels of reactive oxygen species, mitochondrial dysmorphology, and behavioral defects in flies with low levels of Pink1. These findings suggest that Cdk8 regulates Drp1-mediated mitochondrial fission in a similar manner as Pink1 and may contribute to the development of Parkinson’s disease. - Proof-of-Concept Studies With a Computationally Designed Mpro Inhibitor as a Synergistic Combination Regimen Alternative to Paxlovid
As the spread and evolution of SARS-CoV-2 continues, it is important to continue to not only work to prevent transmission but to develop improved antiviral treatments as well. The SARS-CoV-2 main protease (Mpro) has been established as a prominent druggable target. In the current study, investigators evaluate Mpro61 as a lead compound, utilizing structural studies, in vitro pharmacological profiling to examine possible off-target effects and toxicity, cellular studies, and testing in a male and female mouse model for SARS-CoV-2 infection. Results indicate favorable pharmacological properties, efficacy, and drug synergy, as well as complete recovery from subsequent challenge by SARS-CoV-2, establishing Mpro61 as a promising potential preclinical candidate. - Pigs in Transplantation Research and Their Potential as Sources of Organs in Clinical Xenotransplantation
The pig has now gained importance as a potential source of organs for clinical xenotransplantation. When an organ from a wild-type (i.e., genetically unmodified) pig is transplanted into an immunosuppressed nonhuman primate, a vigorous host immune response causes hyperacute rejection (within minutes or hours). This response has been largely overcome by (1) extensive gene editing of the organ-source pig and (2) the administration to the recipient of novel immunosuppressive therapy based on blockade of the CD40/CD154 T-cell costimulation pathway. The combination of gene editing and novel immunosuppressive therapy has extended life-supporting pig kidney graft survival to greater than 1 year and of pig heart survival to up to 9 months. This review briefly describes the techniques of gene editing, the potential risks of transfer of porcine endogenous retroviruses with the organ, and the need for breeding and housing of donor pigs under biosecure conditions. - Trade-Offs Shaping Transmission of Sylvatic Dengue and Zika Viruses in Monkey Hosts
Mosquito-borne dengue (DENV) and Zika (ZIKV) viruses originated in Old World sylvatic (forest) cycles involving monkeys and canopy-living Aedes mosquitoes. Both viruses spilled over into human transmission and were translocated to the Americas, opening a path for spillback into neotropical sylvatic cycles. This article reports that the trade-offs that shape within-host dynamics and transmission of these viruses are lacking, hampering efforts to predict spillover and spillback. The data revealed evidence of an immunologically mediated trade‑off between duration and magnitude of virus replication, as higher-peak ZIKV titers are associated with shorter durations of viremia, and higher natural killer cell levels are associated with lower peak ZIKV titers and lower anti-DENV-2 antibody levels. Furthermore, patterns of transmission of each virus from a neotropical monkey suggest that ZIKV has greater potential than DENV-2 to establish a sylvatic transmission cycle in the Americas. - TGF-Β Blockade Drives a Transitional Effector Phenotype in T Cells Reversing SIV Latency and Decreasing SIV Reservoirs In Vivo
Interruption of antiretroviral therapy leads to rapid rebound of viremia due to the establishment of a persistent viral reservoir early after infection. Using a treatment regimen similar to the one tested in clinical trials, the authors show how galunisertib affects immune cell function, increases simian immunodeficiency virus (SIV) reactivation, and reduces the viral reservoir in female rhesus macaques. Their findings reveal a galunisertib-driven shift toward an effector phenotype in T and natural killer cells. Taken together, this work demonstrates that galunisertib, a clinical-stage TGF-β inhibitor, reverses SIV latency and decreases SIV reservoirs by driving T cells toward an effector phenotype, enhancing immune responses in vivo in the absence of toxicity. - Lymphoid Tissues Contribute to Plasma Viral Clonotypes Early After Antiretroviral Therapy Interruption in SIV-Infected Rhesus Macaques
Researchers are interested in better understanding the sources, timing, and mechanisms of HIV rebound that occurs after interruption of antiretroviral therapy (ART). Using rhesus macaques (sex not specified), investigators tracked barcoded simian immunodeficiency virus (SIV) clonotypes over time and among tissues. Among the tissues studied, mesenteric lymph nodes, inguinal lymph nodes, and spleen contained viral barcodes detected in plasma. Additionally, the authors reported that CD4+ T cells harbored the most viral RNA after ART interruption. These tissues are likely to contribute to viral reactivation and rebound after ART interruption, but further studies are needed to evaluate the relative potential contributions from other tissues and organs. - IL-21-IgFc Immunotherapy Alters Transcriptional Landscape of Lymph Node Cells Leading to Enhanced Flu Vaccine Response in Aging and SIV Infection
Aging is associated with increased risk of seasonal flu disease burden and serious flu-related complications, particularly for people with HIV. In this study, investigators aimed to elucidate the immunomodulation following flu vaccination in aging male and female rhesus macaques infected with simian immunodeficiency virus (SIV). Their results suggest that IL-21 treatment at the time of flu vaccination modulates the inductive lymph node germinal center activity to reverse SIV-associated immune dysfunction. The authors identified IL-21 as a potential candidate molecule for immunotherapy to enhance flu vaccine responses in affected populations. Further studies could examine the overall benefit of IL-21 immunotherapy on mucosal lung immunity and protection against infection. - Validity of Xiphophorus Fish as Models for Human Disease
Xiphophorus is one of the oldest animal systems for studying melanoma. In this article, the authors summarize current Xiphophorus models for other human diseases. They review how Xiphophorus fishes and their interspecies hybrids can be used for studying human diseases and highlight research opportunities enabled by these unique models (both established and emerging). They identified several emerging Xiphophorus models, including for albinism, micromelanophore pigmentation, fin regeneration, and diet-induced obesity. The research on cancer and reproductive maturation discussed in this review substantiates the value of Xiphophorus as a model for human disease throughout all three phases of validation—face, construct, and predictive—and continues to provide important scientific insights. - Identification of Constrained Sequence Elements Across 239 Primate Genomes
Functional genomic elements that have acquired selective constraints specific to the primate order are prime candidates for understanding evolutionary changes in humans, but the selective constraints specific to the phylogenetic branch from which the human species ultimately emerged remain largely unidentified. Researchers constructed a genome-wide multiple sequence alignment of 239 primate species to better characterize constraint at noncoding regulatory sequences in the human genome. Their work reveals noncoding regulatory elements that are under selective constraint in primates but not in other placental mammals and are enriched for variants that affect human gene expression and complex traits in diseases. These findings highlight the important role of recent evolution in regulatory sequence elements differentiating primates, including humans, from other placental mammals. - Injury-Induced Cooperation of InhibinβA and JunB Is Essential for Cell Proliferation in Xenopus Tadpole Tail Regeneration
Certain animal species (e.g., amphibians) that can regenerate lost tissues and limbs after injury offer potential for applications in regenerative medicine. Cell proliferation is essential for the reconstruction of injured tissue, but the molecular mechanisms that regulate the transition from wound healing to regenerative cell proliferation remain unclear. Using Xenopus tropicalis, investigators examined the effects of injury on the expression of inhibin subunit beta A (inhba) and junb. Their findings shed light on the mechanisms underlying injury-induced cell proliferation in regenerative animals. - A Single-Cell Time-Lapse of Mouse Prenatal Development From Gastrula to Birth
In this study, investigators combined single-cell transcriptome profiling of male and female mouse embryos and newborn pups with previously published data to construct a tree of cell-type relationships tracing development from zygote to birth. They applied optimized single-cell combinatorial indexing to profile the transcriptional states of 12.4 million nuclei from 83 embryos, precisely staged at 2- to 6-hour intervals spanning late gastrulation to birth; establish a global framework for exploring mammalian development; and construct a rooted tree of cell-type relationships, from zygote to birth. Their analysis allowed them to systematically nominate genes that encode transcription factors and other proteins as candidate drivers of the in vivo differentiation of hundreds of cell types. Extending this framework to postnatal time points could yield a single-cell time-lapse of the entire mammalian life span, from conception to death. - GenomeMUSter Mouse Genetic Variation Service Enables Multitrait, Multipopulation Data Integration and Analysis
Advances in genetics, including transcriptome-wide and phenome-wide association analysis methods, create compelling new opportunities for using fully reproducible and widely studied inbred mouse strains to characterize the polygenetic basis for individual differences in disease-related traits. Investigators developed an imputation approach and implemented data service to provide a broad and more comprehensive mouse variant resource. They evaluated the strain-specific imputation accuracy on a “held-out” test set that was not used in the imputation process. The authors present its application to multipopulation and multispecies analyses of complex trait variation in type 2 diabetes and substance use disorders and compare these results to human genetics studies. - Interferon Regulatory Factor 7 Modulates Virus Clearance and Immune Responses to Alphavirus Encephalomyelitis
Interferon (IFN) regulatory factor 7 (IRF7)–deficient mice develop fatal paralysis after central nervous system infection with Sindbis virus, whereas wild-type mice recover. Irf7-/- mice produce low levels of IFN-α but high levels of IFN-β with induction of IFN stimulated genes, so the reason for this difference is not understood. The current study shows that Irf7-/- mice developed inflammation earlier but failed to clear virus from motor neuron–rich regions of the brainstem and spinal cord. Therefore, IRF7 is either necessary for the neuronal response to currently identified mediators of clearance or enables the production of additional antiviral factor(s) needed for clearance. - Timing of Initiation of Anti-Retroviral Therapy Predicts Post-Treatment Control of SIV Replication
Researchers are interested in approaches to reducing viral rebound following interruption of antiretroviral therapy, but more work is needed to understand major factors that determine the viral “setpoint” level. Researchers previously assessed how timing of treatment can affect the frequency of rebound from latency. In the current study, the authors analyzed data from multiple studies of simian immunodeficiency virus (SIV) infection in rhesus macaques to further explore the dynamics and predictors of post-treatment viral control. They determined that the timing of treatment initiation was a major predictor of both the level and the duration of post-rebound SIV control. These findings could help inform future treatments. - CD8+ T Cells Control SIV Infection Using Both Cytolytic Effects and Non-Cytolytic Suppression of Virus Production
HIV continuously evades and subdues the host immune responses through multiple strategies, and an understanding of these strategies can help inform research efforts. Using a mathematical model, investigators assessed whether CD8+ cells from male rhesus macaques exert a cytolytic response against infected cells prior to viral production. Their goal was to elucidate the possible mode of action of CD8+ cells on simian immunodeficiency virus (SIV)–infected cells. Models that included non cytolytic reduction of viral production best explained the viral profiles across all macaques, but some of the best models also included cytolytic mechanisms. These results suggest that viral control is best explained by the combination of cytolytic and non-cytolytic effects. - Allelic Strengths of Encephalopathy-Associated UBA5 Variants Correlate Between In Vivo and In Vitro Assays
The UBA5 gene is associated with developmental and epileptic encephalopathy 44 (DEE44), an autosomal recessive disorder, in humans. The link between UBA5 variants and severity of DEE44, however, is not established. Investigators developed humanized fly models carrying a series of patient UBA5 variants. These flies showed differences in survival rates, developmental progress, life span, and neurological well-being. The severity of these defects correlated strongly with functional defects of UBA5 variants, allowing the classification of UBA5 loss-of-function variants into mild, intermediate, and severe allelic strengths in patients. This study provides resources for systematic investigation of the mechanistic link between UBA5 variants and DEE44 and for developing diagnostic approaches. - Body Stiffness Is a Mechanical Property That Facilitates Contact-Mediated Mate Recognition in Caenorhabditis elegans
Body stiffness is a mechanical property that facilitates contact-mediated mate recognition in Caenorhabditis elegans. Chemical cues have been extensively studied as sensory cures of mate recognition, whereas the role of mechanical cues is largely unknown. Investigators studied the link of the hypodermis and body stiffness with mate recognition and mating efficiency in the worm C. elegans. They found that worm males assess attractiveness of potential mates though contact-mediated cues determined by species, sex, and developmental stages of the hypodermis. Body stiffness maintained by a group of cuticular collagens is critical for mate recognition and mating efficiency. This study suggests the important role of mechanosensory cues in mate recognition and provides a platform for mechanistically studying social behavior. - DAZL Knockout Pigs as Recipients for Spermatogonial Stem Cell Transplantation
Spermatogonial stem cell (SSC) transplantation is a technique that holds potential for addressing male infertility, as well as generation of genetically modified animal models. DAZL (Deleted in Azoospermia–Like) is a conserved RNA-binding protein important for germ cell development, and DAZL knockout (KO) causes defects in germ cell commitment and differentiation. Investigators characterized DAZL-KO pigs as SSC transplantation recipients. DAZL-KO pigs support donor-derived spermatogenesis following SSC transplantation, but low spermatogenic efficiency currently limits its use for offspring production. - Biphasic Decay of Intact SHIV Genomes Following Initiation of Antiretroviral Therapy Complicates Analysis of Interventions Targeting the Reservoir
The latent HIV-1 reservoir persists with antiretroviral therapy (ART), and assays for quantifying intact proviruses in nonhuman primate models are needed. Researchers used a simian–human immunodeficiency virus (SHIV) intact proviral DNA assay to describe viral decay during the first year of ART in female rhesus macaques. Their results suggest that intact SHIV genomes in circulating CD4+ T cells undergo biphasic decay during the first year of ART, with a rapid first phase and a slower second phase. These findings provide insight for future studies using SHIV models, as well as new cure interventions. - AZD5582 Plus SIV-Specific Antibodies Reduce Lymph Node Viral Reservoirs in Antiretroviral Therapy–Suppressed Macaques
Researchers are interested in targeting the HIV reservoir via a latency reversal and clearance approach. Previously, investigators demonstrated that AZD5582 induces systemic latency reversal in rhesus macaques and humanized mice, but a consistent reduction in the viral reservoir was not observed. In the current study, they combined AZD5582 with four simian immunodeficiency virus (SIV)–specific rhesus monoclonal antibodies using rhesus macaques of both sexes. They reported a reduction in total and replication-competent SIV DNA in lymph node–derived CD4+ T cells in the treated macaques. These findings provide proof of concept for the potential of the latency reversal and clearance HIV cure strategy. - Intravenous Bacille Calmette–Guérin Vaccination Protects Simian Immunodeficiency Virus–Infected Macaques From Tuberculosis
People with HIV are susceptible to developing tuberculosis and experiencing associated complications. Researchers assessed the safety, immunogenicity, and efficacy of intravenous Bacille Calmette–Guérin vaccination in male and female cynomolgus macaques coinfected with simian immunodeficiency virus (SIV) and Mycobacterium tuberculosis. The vaccine conferred protection in all vaccinated SIV-naïve animals and in 9 of 12 vaccinated SIV-infected animals. These data suggest that the vaccine is immunogenic and efficacious in SIV-infected animals. Overall, this work establishes a model to identify correlates of protection, and these findings can be applied in future studies to develop effective vaccine regimens for people with HIV. - CD8+ Cells and Small Viral Reservoirs Facilitate Post-ART Control of SIV Replication in M3+ Mauritian Cynomolgus Macaques Initiated on ART Two Weeks Post-Infection
A rare group of people infected with HIV can achieve sustainable HIV remission after antiretroviral therapy (ART) withdrawal, but the underlying mechanisms are not fully understood. A team of investigators observed post-treatment control in a cohort of male cynomolgus macaques that were initiated on ART 2 weeks post-infection. Additionally, they reported that the cynomolgus macaques had smaller acute reservoirs than similarly infected rhesus macaques. Collectively, these data suggest that a combination of small reservoirs and immune-mediated virus suppression contributes to post-treatment control in cynomolgus macaques. This model could be used in future studies to develop therapeutic interventions. - A Defect in Mitochondrial Fatty Acid Synthesis Impairs Iron Metabolism and Causes Elevated Ceramide Levels
Human mitochondrial enoyl coenzyme A reductase (Mecr), required for the last step of mitochondrial fatty acid synthesis (mtFAS), is linked to pediatric-onset neurodegeneration, but with unknown mechanisms. Researchers investigated phenotypes of mecr mutants in Drosophila and human-derived fibroblasts. They found that loss of function of Mecr in the whole body resulted in a defect in Fe-S cluster biogenesis and increased iron levels, leading to elevated ceramide levels and lethality in flies. Similar elevated ceramide levels and impaired iron homeostasis were observed in human-derived fibroblasts with Mecr deficiency. Neuronal loss of Mecr led to progressive neurodegeneration in flies. This study pointed out a mechanistic link between mtFAS and neurodegeneration through Mecr. - Spontaneous HIV Expression During Suppressive ART Is Associated With the Magnitude and Function of HIV-Specific CD4+ and CD8+ T Cells
CD4+ and CD8+ T cells are essential in the control of simian immunodeficiency virus and HIV infections, but the mechanisms are not fully understood. Using multiplexed single-cell RNAflow-fluorescence in situ hybridization, researchers quantified and phenotyped viral reservoirs spontaneously expressing viral RNA and the p24 protein in primary clinical samples from men. They reported associations between active reservoirs and HIV-specific CD4+ and CD8+ T cells, and the active reservoirs were largely dominated by defective proviruses. Their findings suggest that viral reservoirs maintain HIV-specific responses during suppressive antiretroviral therapy (ART), and low-level viral gene expression by spontaneous reservoirs is sufficient to maintain anti-HIV adaptive immunity. - Intradermal but Not Intramuscular Modified Vaccinia Ankara Immunizations Protect Against Intravaginal Tier2 Simian–Human Immunodeficiency Virus Challenges in Female Macaques
Researchers have been exploring multiple strategies to develop an HIV vaccine. In this study, the investigators determined the immunogenicity and efficacy of intradermal and intramuscular routes of modified vaccinia Ankara (MVA) vaccination in female rhesus macaques. They found that both routes of MVA vaccination enabled control of viral replication, but only the intradermal vaccination was effective in protection against viral acquisition. Their findings suggest that the intradermal MVA vaccinations provide protection by modulating the innate and T helper responses. This work shows the importance of testing the route of immunization for HIV vaccines in humans. - Proteomic Profiling of Extracellular Vesicles Isolated From Plasma and Peritoneal Exudate in Mice Induced by Crotalus scutulatus Crude Venom and Its Purified Cysteine-Rich Secretory Protein (Css-CRiSP)
Toxins in viperid snakes can induce clinically heterogeneous effects, but most viper venoms are composed of only 10 main protein families. Researchers investigated the proteome expression profile of extracellular vesicles isolated from biofluid samples from male and female mice after injection with crude venom and cysteine-rich secretory proteins. They reported changes in the expression of proteins involved in cell adhesion, cytoskeleton rearrangement, signal transduction, immune responses, and vesicle-mediated transports. This work could be applied in future efforts for early detection and assessment of local effects. - Assessment of Various Standard Fish Diets on Gut Microbiome of Platyfish Xiphophorus maculatus
Diet is an important factor affecting experimental reproducibility and data integration across studies. Reference diets for nontraditional animal models are needed to control diet-induced variation. In a study of the dietary impacts on gut microbiome, researchers found that switching from a custom diet to zebrafish diets altered the platyfish gut microbiome. Their findings suggest that diets developed specifically for zebrafish can affect gut microbiome composition and might not be optimal for platyfish. - A Global Catalog of Whole-Genome Diversity from 233 Primate Species
Researchers are interested in studying the remarkable diversity of morphology and behavior across primates to answer long-standing questions in evolutionary and conservation biology. Using whole-genome sequencing, the investigators created a nuclear DNA phylogeny and reassessed evolutionary divergence times among primate clades. They found that intraspecies genetic diversity across families and geographic regions was associated with climate and sociality but not with extinction risk. Furthermore, mutation rates differed among species, potentially influenced by effective population sizes. Lastly, they identified extensive recurrence of missense mutations that previously were thought to be human specific. - Complement Contributes to Antibody-Mediated Protection Against Repeated SHIV Challenge
The first clinical efficacy trials of a broadly neutralizing antibody (bNAb) resulted in less benefit than expected and suggested that improvements are needed to prevent HIV infection. Using rhesus macaques of both sexes, investigators sought to further investigate the contribution of antibody-mediated activation of complement to the protective potency of an HIV bNAb in passive transfer and simian–human immunodeficiency virus (SHIV) challenge experiments. They observed that fewer bNAbs were required to protect animals from plasma viremia when complement activity was enhanced, suggesting that the functions of complement-mediated effector contribute to in vivo antiviral activity and might contribute to further improvements in the efficacy of antibody-mediated prevention strategies. - An E1–E2 Fusion Protein Primes Antiviral Immune Signaling in Bacteria
Bacteria encode evolutionary predecessors of cGAS called cGAS/DncV-like nucleotidyltransferases (CD-NTases), which detect bacteriophage infection and produce diverse nucleotide second messengers, but how bacterial CD-NTase activation is controlled remains unknown. Investigators showed that the cGAS/DncV–like nucleotidyltransferase (CD NTase)–associated protein 2 (Cap2) primes bacterial CD-NTases for activation through a ubiquitin transferase–like mechanism. The study also shows that a specific endopeptidase, Cap3, balances Cap2 activity by cleaving CD-NTase-target conjugates. These findings demonstrate that bacteria control immune signaling using an ancient, minimized ubiquitin transferase–like system and provide insight into the evolution of the E1 and E2 machinery across domains of life. - Spike and Nsp6 Are Key Determinants of SARS-CoV-2 Omicron BA.1 Attenuation
The ability of the SARS-CoV-2 virus to mutate and create variants of concern demands new vaccines to control the COVID-19 pandemic. The SARS-CoV-2 Omicron variant was shown to be more immune evasive and less virulent than current major variants. The spike (S) protein in this variant carries many mutations that drive these phenotypes. Researchers generated a chimeric recombinant SARS-CoV-2 virus encoding the S gene of Omicron (BA.1 lineage) in an ancestral SARS-CoV-2 isolate and compared it with the naturally circulating Omicron variant. The Omicron S-bearing virus escaped vaccine-induced humoral immunity, owing to mutations in the receptor-binding motif. The recombinant virus replicated efficiently in distal lung cell lines and in K18-hACE2 mice. Moreover, mutations induced in non-structural protein 6 (nsp6) in addition to the S protein were sufficient to restate the attenuated phenotype of Omicron. These findings indicate that the pathogenicity of Omicron is determined by mutations both inside and outside of the S gene. - Giardia Hinders Growth by Disrupting Nutrient Metabolism Independent of Inflammatory Enteropathy
Giardia lamblia is one of the most common intestinal pathogens among children in low- and middle-income countries. Investigators performed translational investigations using the Malnutrition and Enteric Diseases (MAL-ED) male and female cohort, as well as mice of both sexes, to identify mechanistic pathways that might explain Giardia-induced effects on early childhood growth. They identified signatures in the urinary metabolome of young children, suggesting that host growth restriction during infection is mediated by dysregulated amino acid metabolism. - Mechanism of STMN2 Cryptic Splice-Polyadenylation and Its Correction for TDP-43 Proteinopathies
Loss of the RNA-binding protein TDP-43 from the nuclei of affected neurons is a hallmark of neurodegeneration in TDP-43 proteinopathies (e.g., amyotrophic lateral sclerosis, frontotemporal dementia). Loss of functional TDP-43 is accompanied by misprocessing of the stathmin-2 (STMN2) RNA precursor. Investigators determined the elements through which TDP-43 regulates STMN2 pre mRNA processing and identified steric binding antisense oligonucleotides that are capable of restoring normal STMN2 protein and RNA levels. This approach could be applied to the development of new therapies. - The Landscape of Tolerated Genetic Variation in Humans and Primates
Investigators created a whole-genome sequence database from 809 individual nonhuman primates (NHPs) of 233 different primate species to test the hypothesis that gene variants that do not cause disease in NHPs also would likely be benign in humans. They found that 99% of the genetic variants that were benign in NHPs also were classified as benign in the human ClinVar database. In contrast, only 71% to 87% of genomic variants classified as benign in nonprimate animals were benign in humans. Building on this approach, the authors reclassified more than 4 million human genetic variants of unknown health impact as likely being benign based on effects in NHPs. This work illustrates the power of comparative medicine approaches between nonhuman primates and humans. - Chronic Immune Activation and Gut Barrier Dysfunction Is Associated with Neuroinflammation in ART-Suppressed SIV+ Rhesus Macaques
About 40% of people with HIV develop neurocognitive disorders, potentially resulting from persistent infection in the brain and neuroinflammation. Investigators characterized the central nervous system reservoir and immune environment of simian immunodeficiency virus (SIV)–infected rhesus macaques of both sexes during acute, chronic, or antiretroviral therapy (ART)–suppressed infection. They reported that neuroinflammation and blood–brain barrier dysfunction correlated with viremia and immune activation in the gut. Their findings suggest that gastrointestinal tract damage can contribute to neuroimmune activation and inflammation, even in the absence of SIV or HIV infection. This work also has implications for other neurological disorders where chronic inflammation is associated with pathogenesis. - Cannabinoids Modulate the Microbiota–Gut–Brain Axis in HIV/SIV Infection by Reducing Neuroinflammation and Dysbiosis while Concurrently Elevating Endocannabinoid and Indole‑3‑Propionate Levels
Chronic neuroinflammation is thought to be a significant contributor to HIV-associated neurocognitive disorders. Using rhesus macaques of both sexes, researchers investigated the effects of simian immunodeficiency virus (SIV) infection on the microbiota–gut–brain axis (MGBA), as well as the use of low-dose cannabinoids to reverse MGBA dysregulation. They reported that tetrahydrocannabinol reduced neuroinflammation and dysbiosis and increased plasma endocannabinoid, endocannabinoid-like, glycerophospholipid, and indole-3-propionate levels. This study offers a potential strategy to promote brain health in people with HIV. - Neuroprotective Effects of Electrical Stimulation Following Ischemic Stroke in Non-Human Primates
Using rhesus macaques of both sexes, researchers identified a novel treatment for ischemic stroke, which occurs when brain cells die due to lack of oxygen. The treatment consisted of applying 60 minutes of electrical brain stimulation shortly after the stroke. The animals that received electrical stimulation had less brain damage, fewer cell deaths, and more protective neural activity patterns than the monkeys that did not receive electrical stimulation. Future work can determine whether this stimulation can be applied noninvasively, as well as how to improve the electrical stimulation patterns to optimize health outcomes for stroke patients. - High Resolution Genomes of Multiple Xiphophorus Species Provide New Insights into Microevolution, Hybrid Incompatibility, and Epistasis
Existing Xiphophorus genome assemblies are not at the chromosomal level and are prone to sequence gaps, hindering advancement of evolutionary, comparative, and translational biomedical studies. Investigators assembled high-quality chromosome-level genome assemblies for three distantly related Xiphophorus species. They found that expanded gene families and positively selected genes associated with live bearing. Positively selected gene families were enriched in nonpolymorphic transposable elements, suggesting that dispersal has accompanied the evolution of the genes, possibly by incorporating new regulatory elements. The investigators also characterized interspecific polymorphisms, structural variants, and polymorphic transposable element insertions and assessed their association to interspecies hybridization-induced gene expression dysregulation related to specific disease states in humans. - Prolonged Experimental CD4+ T-Cell Depletion Does Not Cause Disease Progression in SIV-Infected African Green Monkeys
African green monkeys (AGMs) chronically infected with simian immunodeficiency virus (SIV) partially recover mucosal CD4+ T cells, maintain gut integrity, and do not progress to AIDS. Investigators assessed the impact of prolonged, antibody-mediated CD4+ T cell depletion on gut integrity and natural history of SIV infection in AGMs. All circulating CD4+ T cells and more than 90% of mucosal CD4+ T cells were depleted. Plasma viral loads and cell-associated viral RNA in tissues were lower in CD4+ cell–depleted animals. CD4+ cell–depleted AGMs maintained gut integrity, controlled immune activation, and did not progress to AIDS. Therefore, CD4+ T cell depletion is not a determinant of SIV-related gut dysfunction when gastrointestinal tract epithelial damage and inflammation are absent, suggesting that disease progression and resistance to AIDS are independent of CD4+ T cell restoration in SIV-infected AGMs. - Surrogate Biomarkers of Disease Progression in Human Pegivirus–Seropositive Human Immunodeficiency Virus–Infected Individuals
Researchers have previously observed that human pegivirus (HPgV) infection is associated with reduced progression of HIV. Investigators examined markers of HIV progression in male and female individuals with HIV and HPgV infection. They reported that HIV plasma viral load was lower in HPgV-seropositive individuals with HIV than in HPgV‑seronegative individuals with HIV. They also found that clinical markers of hepatic damage were significantly lower in HPgV-seropositive individuals with HIV. Future work could examine pathways through which HPgV influences HIV control, which might inform the development of new therapeutics. - Impaired Placental Hemodynamics and Function in a Non-Human Primate Model of Gestational Protein Restriction
Maternal malnutrition is a global health epidemic that adversely affects fetal outcomes and results in long-term health complications in children. Investigators used a previously developed model in nonhuman primates for gestational protein restriction to study the impact of undernutrition, specifically protein deficiency, on placental function and pregnancy outcomes. The data demonstrate that a 50% protein-restricted diet reduces maternal placental perfusion, decreases fetal oxygen availability, and increases fetal mortality. These alterations in placental hemodynamics could partly explain human growth restriction and stillbirth seen with severe protein restriction in developing countries. - Infant Rhesus Macaques Immunized Against SARS-CoV-2 Are Protected Against Heterologous Virus Challenge 1 Year Later
The Moderna and Pfizer–BioNTech mRNA vaccines received emergency-use authorization for infants 6 months and older in June 2022, but questions remain regarding the durability of vaccine efficacy against emerging variants in this age group. Using a two-dose vaccine regimen consisting of stabilized prefusion Washington-strain spike protein encoded by mRNA and encapsulated in lipid nanoparticles, the investigators immunized 2-month-old rhesus macaques of both sexes. They found that the immune responses persisted and protected them from severe disease after heterologous challenge with the Delta variant 1 year later. The decay kinetics of vaccine-induced neutralizing antibody responses in the infant monkeys are comparable to those observed in adult humans and nonhuman primates. - Molecular Insights into Antibody-Mediated Protection Against the Prototypic Simian Immunodeficiency Virus
Most simian immunodeficiency virus (SIV) vaccines have focused on inducing T cell responses alone or in combination with non-neutralizing antibody responses. To date, studies investigating neutralizing antibody (nAb) responses to protect against SIV have been limited. In this study, researchers isolated 12 potent monoclonal nAbs from chronically infected rhesus macaques of both sexes and mapped their binding specificities on the envelope trimer structure. They further characterized the structures using cryogenic electron microscopy, mass spectrometry, and computational modeling. Their findings indicate that, in the case of humoral immunity, nAb activity is necessary and sufficient for protection against SIV challenge. This work provides structural insights for future vaccine design. - A Multidimensional Metabolomics Workflow to Image Biodistribution and Evaluate Pharmacodynamics in Adult Zebrafish
The evaluation of tissue distribution and pharmacodynamic properties of a drug is essential but often expensive in clinical research. The investigators developed a multidimensional metabolomics platform to evaluate drug activity that integrates mass spectrometry–based imaging, absolute drug quantitation, in vivo isotope tracing, and global metabolome analysis in zebrafish. They validated this platform by evaluating whole-body distribution of the anti-rheumatic agent hydroxychloroquine sulfate and its impact on the systemic metabolism of adult zebrafish. This work suggests that the multidimensional metabolomics platform is a cost-effective method for evaluating on- and off-target effects of drugs. - Two Neuronal Peptides Encoded from a Single Transcript Regulate Mitochondrial Complex III in Drosophila
Transcripts with small open-reading frames (smORFs) are underrepresented in genome annotations. Functions of peptides encoded by smORFs are poorly understood. The investigators systematically characterized human-conserved smORF genes in Drosophila and found two peptides, Sloth1 and Sloth2, that are highly expressed in neurons. They showed that Sloth1 and Sloth2 are paralogs with high sequence similarity but are not functionally redundant. Loss of either peptide resulted in lethality, neurodegeneration, and impaired mitochondrial function. This work suggests the value of phenotypic analysis of smORFs using Drosophila as a model. - Rapid Joule Heating Improves Vitrification-Based Cryopreservation
Cryopreservation by vitrification is an effective approach for long-term preservation of biosystems, but effective vitrification often requires high concentrations of cryoprotective agent (CPA), which can be toxic. The investigators described a joule heating–based platform technology for rapid rewarming of biosystems, which allows the use of low concentrations of CPA. They demonstrated the success of this platform in cryopreservation with low CPA concentrations in three model systems: adherent cells, Drosophila melanogaster embryos, and rat kidney slices. This work provides a general solution to cryopreserve a broad spectrum of cells, tissues, organs, and even whole animals. - TMEM161B Modulates Radial Glial Scaffolding in Neocortical Development
Neocortical folding (i.e., gyrification) is a fundamental evolutionary mechanism allowing the expansion of cortical surface area and increased cognitive function. This study identifies TMEM161B, a transmembrane protein, in gyral spacing in humans, likely affecting radial glial cell polarity through effects on the actin cytoskeleton. Patients carrying TMEM161B mutations exhibit striking neocortical polymicrogyria and intellectual disability. TMEM161B knockout mice fail to develop midline hemispheric cleavage, whereas knock-in of patient mutations and patient-derived brain organoids show defects in apical cell polarity and radial glial scaffolding. The data implicating TMEM161B in murine holoprosencephaly may suggest shared mechanisms between the formation of the brain midline and cortical gyrification. - Genome Structures Resolve the Early Diversification of Teleost Fishes
The early evolution of teleost fishes remains an unanswered question among evolutionary biologists. The three earliest branching clades of crown teleosts are Elopomorpha (e.g., tarpons, eels), Osteoglossomorpha (e.g., arapaima, elephantnose fish), and Clupeocephala (e.g., zebrafish, medaka). Building on recently described genome assemblies in Elopomorpha, the authors explored teleost phylogeny using independent gene sequencing and chromosomal rearrangement phylogenomic approaches. They found that Elopomorpha and Osteoglossomorpha comprise a monophyletic sister group to all other teleosts. This report highlights the value of combining different levels of genome-wide information to solve complex phylogenies and will serve as a basis for new investigations into the genomic and functional evolution of teleosts. - Isoniazid and Rifapentine Treatment Effectively Reduces Persistent M. tuberculosis Infection in Macaque Lungs
People with HIV and asymptomatic latent tuberculosis (TB) coinfection are at risk of developing active TB symptoms. The Centers for Disease Control and Prevention recommends a weekly dose of isoniazid and rifapentine for 3 months (3HP) for treatment of latent TB infection, but the sterilizing efficacy of the regimen has not been demonstrated previously. Using rhesus macaques of both sexes, researchers evaluated the efficacy of the 3HP regimen in eradicating persistent Mycobacterium tuberculosis infection. They found that treatment reduced the risk of developing active TB but did not establish complete sterilization. This work establishes a new animal model for evaluating the efficacy of different drug regimens. - A Clade C HIV-1 Vaccine Protects Against Heterologous SHIV Infection By Modulating IgG Glycosylation and T Helper Response in Macaques
Vaccines for HIV-1 capable of generating a broadly cross-reactive neutralizing antibody response are needed urgently. The researchers tested the protective efficacy of a clade C HIV-1 vaccination regimen in male rhesus macaques. The vaccine was administered either orally using a needle-free injector or through parenteral injection. Significant protection was observed for both vaccination routes following the simian–human immunodeficiency virus (SHIV) challenge, with an estimated efficacy of 68% per exposure. These results suggest that glycosylation profile of IgG and HIV-resistant helper T cell response contributes to vaccine protection. - Neuroinflammatory Transcriptional Programs Induced in Rhesus Pre‑Frontal Cortex White Matter During Acute SHIV Infection
Neuroinflammation has evolved as a protective immune response within the central nervous system (CNS), but chronic neuroinflammation leads to oxidative stress, cellular damage, and neurodegeneration. People with HIV are at increased risk for age-related neurodegenerative diseases. Using rhesus macaques of both sexes, the researchers characterized the molecular underpinnings of acute neuroinflammation following simian–human immunodeficiency virus (SHIV) infection. Viral entry and integration within the CNS demonstrated vulnerabilities of key cognitive and motor function brain regions during the acute phase of infection. SHIV-induced transcriptional alterations also were observed. These findings indicate the presence of pervasive immune surveillance at homeostasis and reveal key perturbations during infection. - Metabolic Transitions Define Spermatogonial Stem Cell Maturation
The spermatogonial stem cell (SSC) is the basis of male fertility. One potential option to preserve fertility in patients treated with anti-cancer therapy is isolation and laboratory culture of the juvenile SSC pool with subsequent transplantation to restore spermatogenesis. However, efficient culture of undifferentiated spermatogonia, including SSCs, in mammals other than rodents remains challenging. Investigators reported that the metabolic phenotype of prepubertal human spermatogonia is distinct from that of adult spermatogonia and that SSC development is characterized by specific metabolic transitions from oxidative phosphorylation to anaerobic metabolism. - Promoting Validation and Cross-Phylogenetic Integration in Model Organism Research
Model organisms (MOs) are essential for biomedical research and therapeutic development. However, translation of MO research to the clinic is low. Workshop and session chairs summarized the discussions from a series of NIH-supported workshops, titled “Validation of Animal Models and Tools for Biomedical Research,” that was held in 2020 and 2021. The authors describe challenges and opportunities for developing and integrating tools and resources and provide suggestions for improving the rigor, validation, reproducibility, and translatability of MO research. Finally, the authors point out the importance of these suggested improvements in facilitating the validation of animal models for human biology and disease. - SARS-CoV-2 Infects Neurons and Induces Neuroinflammation in a Non‑human Primate Model of COVID-19
SARS-CoV-2 causes brain fog and other neurological complications in some patients. It has been unclear whether SARS-CoV-2 infects the brain directly or whether central nervous system sequelae result from systemic inflammatory responses triggered in the periphery. Using a rhesus macaque model, researchers detected SARS-CoV-2 in the olfactory cortex and interconnected regions 7 days after infection in both sexes, with predominance of neuronal infection along corticocortical pathways, suggesting that the virus enters the brain through the olfactory nerve. Neuroinflammation and neuronal damage were more severe in elderly monkeys with type 2 diabetes. The researchers found that in aged monkeys, SARS-CoV-2 traveled farther along nerve pathways to regions associated with Alzheimer’s disease. - Parallel Processing, Hierarchical Transformations, and Sensorimotor Associations Along the “Where” Pathway
Visually guided behaviors require the brain to transform ambiguous retinal images into object-level spatial representations and map those representations to motor responses. These capabilities are supported by the dorsal “where” pathway in the brain, but the specific contributions of areas along this pathway have remained elusive. Using male rhesus macaques, the researchers compared neuronal activity in two areas along the “where” pathway that bridge the parieto‑occipital junction: intermediate visual area V3A and the caudal intraparietal (CIP) area. Neuronal activity was recorded while the animals made perceptual decisions based on judging the tilt of 3D visual patterns. The investigators found that CIP shows higher-order spatial representations and more choice-correlated responses, which support a V3A-to-CIP hierarchy. The researchers also discovered modulation of V3A activity by extraretinal factors, suggesting that V3A might be better characterized as contributing to higher-order behavioral functions rather than low-level visual feature processing. - Reduced Alcohol Preference and Intake After Fecal Transplant in Patients with Alcohol Use Disorder Is Transmissible to Germ-free Mice
Alcohol use disorder is a major cause of reduced life expectancy worldwide, and this misuse has increased exponentially during the COVID-19 pandemic. Fecal microbiota transplant has been shown previously to reduce alcohol craving in humans with cirrhosis. Here, the investigators report that the reduction in craving and alcohol preference is transmissible to male germ-free mice only when live bacteria—and not germ-free supernatants—are used for colonization. This differential colonization was associated with alterations in the gut immune–inflammatory response through short-chain fatty acids. - Targeted Suppression of Human IBD-Associated Gut Microbiota Commensals by Phage Consortia for Treatment of Intestinal Inflammation
Human gut commensals increasingly are suggested to affect noncommunicable diseases, such as inflammatory bowel disease (IBD), yet their targeted suppression remains an unmet challenge. In this report, investigators identified a clade of Klebsiella pneumoniae (Kp) strains—featuring a unique antibiotic resistance and mobilome signature—that is associated strongly with disease exacerbation and severity. Transfer of clinical IBD-associated Kp strains into colitis-prone, germ-free, and colonized mice of both sexes enhances intestinal inflammation. Stepwise generation of a lytic five-phage combination, targeting sensitive and resistant IBD-associated Kp clade members, enables effective Kp suppression. These data demonstrate the feasibility of orally administered combination phage therapy in avoiding resistance while effectively inhibiting noncommunicable disease–contributing pathobionts. - Lesion Environments Direct Transplanted Neural Progenitors Towards a Wound Repair Astroglial Phenotype in Mice
Neural progenitor cells (NPCs) are potential cell transplantation therapies for central nervous system (CNS) injuries. Investigators derived NPCs expressing a ribosomal protein-hemagglutinin tag (RiboTag) for transcriptional profiling. Their findings reveal similarities between the transcriptional profiles, cellular morphologies, and certain functional features of cells transplanted into subacute CNS lesions and host astroglia. The astroglia are stimulated by CNS injuries to proliferate and adopt a naturally occurring, border-forming wound repair phenotype in mice of both sexes. Understanding the autonomous cues instructing NPCs transplanted in CNS host tissue will be fundamental to achieving mechanism-based approaches to therapeutic NPC transplantation. - Durable Protection Against the SARS-CoV-2 Omicron Variant Is Induced by an Adjuvanted Subunit Vaccine
Additional SARS-CoV-2 vaccines are needed, owing to waning immunity to the original vaccines and the emergence of variants of concern. A recent study in male rhesus macaques demonstrated durable protection against the Omicron BA.1 variant induced by a subunit SARS-CoV-2 vaccine comprising the receptor‑binding domain of the ancestral strain (RBD-Wu) on the I53-50 nanoparticle adjuvanted with AS03, an oil-in-water emulsion containing α‑tocopherol. Two immunizations with the vaccine resulted in durable immunity, without cross-reactivity. Further boosting with a version of the vaccine containing the Beta variant or the ancestral RBD elicited cross-reactive immune responses that conferred protection against Omicron challenge. - Large Comparative Analyses of Primate Body Site Microbiomes Indicate that the Oral Microbiome Is Unique Among All Body Sites and Conserved Among Nonhuman Primates
Microbiomes are critical to host health and disease, but large gaps remain in our understanding of the determinants, coevolution, and variation of microbiomes across body sites and host species. Thus, researchers conducted the largest comparative study of primate microbiomes to date by investigating microbiome community composition at eight distinct body sites in 17 host species. They found that the oral microbiome is unique in exhibiting notable similarity across primate species while being distinct from the microbiomes of all other body sites and host species. This finding suggests conserved oral microbial niche specialization, despite substantial dietary and phylogenetic differences among primates. - Profiling Development of Abdominal Organs in the Pig
The pig is a model system for studying human development and disease due to its similarities to human anatomy, physiology, size, and genome. Moreover, advances in CRISPR gene editing have made genetically engineered pigs a viable model for the study of human pathologies and congenital anomalies. Investigators describe normal development of the pig abdominal system (i.e., kidney, liver, pancreas, spleen, adrenal glands, bowel, gonads) as an atlas to study congenital defects that can arise in CRISPR-edited Sin3a-associated protein 130 (SAP130) mutant pigs of both sexes. This atlas and the methods described here can be used as tools for identifying developmental pathologies of the abdominal organs in the pig at different stages of development. - Mosaic RBD Nanoparticles Protect Against Challenge by Diverse Sarbecoviruses in Animal Models
Vaccines and therapeutics are needed to protect against both SARS-CoV-2 variants and zoonotic sarbecoviruses with the potential to infect humans. Researchers designed mosaic-8 nanoparticles (SARS-CoV-2 and seven animal sarbecoviruses) that present randomly arranged sarbecovirus spike receptor-binding domains (RBDs) as immunogens to elicit antibodies against epitopes that are conserved and relatively occluded rather than variable, immunodominant, and exposed. Results of immune responses elicited by mosaic-8 RBD nanoparticles in female mice and macaques of both sexes suggest that mosaic nanoparticles could protect against both SARS-CoV-2 variants and zoonotic sarbecoviruses with the potential to infect humans. - Maternal Western-Style Diet Reduces Social Engagement and Increases Idiosyncratic Behavior in Japanese Macaque Offspring
Evidence points to an association between maternal obesity and risk of early-emerging neurodevelopmental disorders in offspring, yet few preclinical studies have tested for associations between maternal Western-style diet (mWSD) and offspring behavior. Using Japanese macaques, researchers found that mWSD offspring of both sexes exhibited less proximity to peers and initiated fewer affiliative social behaviors. These behavioral outcomes appear to be mediated by increased maternal interleukin-12 during the third trimester of pregnancy. Additionally, mWSD offspring displayed increased idiosyncratic behavior, which was related to alterations in maternal adiposity and leptin. These findings suggest specific prevention and intervention targets for early-emerging neurodevelopmental disorder in humans. - Social Connections Predict Brain Structure in a Multidimensional Free-Ranging Primate Society
Reproduction and survival in most primate species reflect management of both competitive and cooperative relationships. The researchers investigated the links between neuroanatomy and sociality in free-ranging rhesus macaques of both sexes. In adults, the number of social partners predicted the volume of the mid‑superior temporal sulcus and ventral-dysgranular insula, which are implicated in social decision-making and empathy, respectively. Scientists found no link between brain structure and other key social variables in adults, such as social status or indirect connectedness, nor between maternal social networks or status and dependent infant brain structure. These findings demonstrate that the size of specific brain structures varies with the number of direct affiliative social connections and suggest that this relationship could arise during development. These results reinforce proposed links among social network size, biological success, and the expansion of specific brain circuits. - Allogeneic MHC‑Matched T‑cell Receptor α/β‑Depleted Bone Marrow Transplants in SHIV‑Infected, ART‑Suppressed Mauritian Cynomolgus Macaques
Allogeneic hematopoietic stem cell transplants are effective in reducing HIV reservoirs following antiretroviral therapy (ART). A better understanding of this mechanism could enable the development of safer and more efficacious HIV treatment regimens. In this study, the researchers used a Mauritian cynomolgus macaque model (sex not specified) to study the effects of allogeneic major histocompatibility complex–matched α/β T cell–depleted bone marrow cell transplantation following infection with simian–human immunodeficiency virus (SHIV). The macaques began ART 6–16 weeks post-infection. In three of the four macaques, SHIV DNA was undetectable in blood but persisted in other tissues. These results suggest that extended ART likely is needed to eradicate the HIV reservoir following transplantation. In future studies, full donor engraftment should be balanced with suppression of graft-versus-host disease. - Innate Immune Regulation in HIV Latency Models
Researchers are interested in developing therapeutic approaches to target latent HIV reservoirs, which are unaffected by antiretroviral therapy. Previous studies suggest that HIV latency might be related to viral RNA sensing, interferon (IFN) signaling, and IFN-stimulated gene (ISG) activation. In this study, the researchers evaluated responses to stimulation by retinoic acid–inducible gene I agonists and IFN in multiple CD4+ T cell line models for HIV latency. The models represented various aspects of latent infection and viral control. Several of the cell lines demonstrated reduced ISG induction, suggesting that long-term latency might be related to dysregulation of the downstream IFN response. These effects likely reflect transcriptional changes occurring within a core set of ISGs and altering IFN responses. Additional studies could provide insight into the functions of these ISGs in HIV latency. - Myeloid Cell Tropism Enables MHC-E–Restricted CD8+ T Cell Priming and Vaccine Efficacy by the RhCMV/SIV Vaccine
Simian immunodeficiency virus (SIV) vaccines based on strain 68-1 rhesus cytomegalovirus vectors have been shown to arrest viral replication early in primary infection. The specific characteristics underlying this effect are not understood fully. In this study, the researchers used host microRNA–mediated vector tropism restriction to demonstrate that the targeted responses are dependent on vector infection of distinct cell types in a rhesus macaque model of both sexes. Only vectors programmed to elicit major histocompatibility complex E–restricted CD8+ T cell responses provided protection against SIV challenge. These findings could be applied in the development of other vaccines for cancers and infectious diseases. - A Cellular Trafficking Signal in the SIV Envelope Protein Cytoplasmic Domain Is Strongly Selected for in Pathogenic Infection
Envelope glycoproteins within the cytoplasmic domain of HIV and simian immunodeficiency virus (SIV) include a tyrosine-based motif that mediates endocytosis and polarized sorting in infected cells. Mutation of this tracking signal has been shown to lead to suppressed viral replication and failed systemic immune activation, but the mechanism has not been explored fully. Using rhesus and pigtail macaque models (sex not specified), the researchers demonstrated that molecular clones containing the mutations reconstitute signals for both endocytosis and polarized sorting. Their findings suggest strong selection pressure for these processes during pathogenic HIV and SIV infection. - CAR/CXCR5–T Cell Immunotherapy Is Safe and Potentially Efficacious in Promoting Sustained Remission of SIV Infection
HIV and simian immunodeficiency virus (SIV) replication are concentrated within the B cell follicles of secondary lymphoid tissues. In this study, the researchers developed immunotherapeutic chimeric antigen receptor (CAR) T cells that home to follicles and clear SIV-infected cells in a rhesus macaque model of both sexes. The CAR T cells localized to the follicle, replicated, and interacted directly with infected cells. Most of the treated animals maintained lower viral loads in the blood and follicles than control animals. These findings demonstrate the safety and potential efficacy of this immunotherapy approach for long-term remission of HIV without requiring the lifelong use of antiretroviral therapy. - A Constitutively Expressed Fluorescent Ubiquitination-Based Cell-Cycle Indicator (FUCCI) in Axolotls for Studying Tissue Regeneration
Regulation of cell-cycle progression is essential for cell proliferation during regeneration post-injury. After appendage amputation, the axolotl, Ambystoma mexicanum, regenerates missing structures through an accumulation of proliferating cells known as the blastema. To study cell division during blastema growth, the authors generated a transgenic line of axolotls that ubiquitously expresses a bicistronic version of the fluorescent ubiquitination-based cell-cycle indicator. This new line of animals will be useful for studying cell-cycle dynamics using in situ endpoint assays and in vivo imaging in developing and regenerating animals. - Gene and Transgenics Nomenclature for the Laboratory Axolotl—Ambystoma mexicanum
The axolotl, Ambystoma mexicanum, is widely used in biological research because of its remarkable capacity for regeneration. Recent advancements in genetic and molecular toolkits are greatly accelerating scientific research using the axolotl, especially regarding tissue regeneration. At this juncture, a critical need exists to establish gene and transgenic nomenclature to ensure uniformity in axolotl research. Here, the authors propose guidelines for unified, genetic nomenclature when working with the axolotl. - A Neutralizing Antibody Target in Early HIV-1 Infection Was Recapitulated in Rhesus Macaques Immunized with the Transmitted/Founder Envelope Sequence
Envelope proteins (Envs) of HIV-1 likely contain features that drive neutralization breadth. Researchers immunized rhesus macaques of both sexes using Envs from HIV-1-infected human subjects who developed either high or low neutralizing antibody (nAb) levels. Several animals developed nAbs when immunized with the Env from the high-neutralizing individual. Investigators mapped the targets of vaccine-elicited nAbs to a distinct loop region of Env, which also was targeted by neutralizing monoclonal antibodies from the infected individual and immunized monkeys. This is a promising starting point to drive nAb breadth, an ultimate goal of vaccine-design efforts. - Chloride Channel Accessory 1 Gene Deficiency Causes Selective Loss of Mucus Production in a New Pig Model
Chloride channel accessory 1 (CLCA1) is thought to be essential for the expression of MUC5AC, an inflammatory mucin, in response to type 2 cytokines. Investigators used a knockout pig model to demonstrate the loss of MUC5AC+ cells throughout the airway mucosa. Consistent effects were observed in the intestinal mucosa. These findings suggest that CLCA1 plays a role in controlling the expression of MUC5AC expression during mucus production. This work provides a new animal model that can be used for further studies of mucus production at respiratory and intestinal sites. - Suppression of Human and Simian Immunodeficiency Virus Replication with the CCR5-Specific Antibody Leronlimab in Two Species
The prevalence of HIV drug resistance has increased in recent years, in large part because of the use of antiretroviral therapy by people with HIV. Researchers showed that injections of leronlimab—an anti-CCR5 antibody that blocks the binding of HIV to CCR5—suppressed HIV replication in five patients with HIV for more than 7 years. They also found that the average amount of virus in the blood of leronlimab-treated rhesus macaques was 10,000 times lower than in untreated animals. These findings suggest that leronlimab is a safe and effective anti-HIV therapeutic drug. - A Basement Membrane Discovery Pipeline Uncovers Network Complexity, Regulators, and Human Disease Associations
Basement membranes are sheet-like networks of extracellular matrix proteins that support and surround tissues. Defects in these membranes are associated with such human conditions as diabetes, kidney disorders, cancer, and cardiovascular issues. Researchers developed an experimental pipeline to unravel the full scale of proteins that make up basement membranes by combining large-scale data analysis, genetic studies, and imaging to track basement membranes in living animals. The study led to the identification of more than 200 human basement membrane proteins and new genetic disease associations, which could lead to better diagnosis and treatment of basement membrane defects in cardiovascular and kidney disorders, fibrosis, and aging. - Voltage Imaging Identifies Spinal Circuits That Modulate Locomotor Adaptation in Zebrafish
Spinal circuits underlying rhythmic locomotion are well described; however, little is known about how the network modulates its output strength, in part due to the difficulty of recording spinal neurons during behavior. The authors used voltage imaging to map the membrane potential of large populations of glutamatergic (V3) neurons throughout the spinal cord of the larval zebrafish during fictive swimming. Optogenetic activation of V3 neurons led to stronger swimming and longer bouts without affecting tail beat frequency. Genetic ablation of V3 neurons led to reduced locomotor adaptation. Based on these findings, V3 neurons appear to be a critical driver of locomotor adaptation in zebrafish. - A Simple Standard for Sharing Ontological Mappings (SSSOM)
Diseases can be represented by identifiers in diverse databases. Merging these databases without accurate mappings, however, might result in loss of important information, resulting in inaccurate or incorrect disease diagnoses. The SSSOM addresses shortcomings in current bioinformatics information mapping schemas. SSSOM is a simple, standardized format that facilitates the sharing of high-quality mappings between different data sources, bringing together biomedical databases and enabling more accurate diagnosis of diseases. In this report, the authors present the SSSOM standard, describe several use cases, and survey current work in this area. - Ceramides Are Early Responders in Metabolic Syndrome Development in Rhesus Monkeys
Metabolic syndrome negatively affects millions of people in the United States. Researchers investigated 16 overweight male rhesus macaques, half of which spontaneously developed metabolic syndrome. Circulating sphingolipids were altered early in the development of insulin resistance, independent of changes in adiposity and before loss of euglycemia. The interactions among circulating sphingolipids and among other insulin-responsive biomolecules—apparent in healthy animals—were lost in the impaired monkeys before the onset of insulin resistance. Thus, lipid profiling can be used to identify early biomarkers of insulin resistance and shed light on the underlying biology of metabolic impairment. - Antithetic Effect of Interferon-α on Cell-Free and Cell-to-Cell HIV-1 Infection
HIV-1 experiences a strong bottleneck during transmission; viruses with higher resistance to host innate immunity tend to be transmitted. Researchers investigated resistance to interferon-alpha (IFN-α)–mediated antiviral effects in HIV-1 isolated at the transmission/founder (TF) and chronic control (CC) phases of infection. IFN-α suppresses cell-to-cell transmission of CC virus but only weakly affects the TF virus. The researchers found that IFN-α enhances cell-free HIV-1 infection, particularly that of CC virus, in a virus-cell density-dependent manner. These results provide insights into how HIV-1 evolves to counteract or hijack host immunity. - Neuropathology and Virus in Brain of SARS-CoV-2-Infected Non-human Primates
Investigators observed neuroinflammation, microhemorrhages, brain hypoxia, and neuropathology that are consistent with hypoxic-ischemic injury in SARS-CoV-2-infected nonhuman primates (NHPs). This effect is seen among infected animals that do not develop severe respiratory disease, which may provide insight into neurological symptoms associated with long COVID-19. Sparse virus was detected in brain endothelial cells but did not associate with the severity of central nervous system injury. These results are indicative of neuropathogenesis of SARS-CoV-2 infection and demonstrate SARS-CoV-2-infected NHPs are a relevant animal model for investigating COVID-19 neuropathogenesis. Read more about the pervasive effects of COVID-19 on the brain of nonhuman primates. - Reduced Infant Rhesus Macaque Growth Rates Due to Environmental Enteric Dysfunction and Association with Histopathology in the Large Intestine
Researchers characterized environmental enteric (relating to the intestines) dysfunction (EED) among infant rhesus macaques (n=80, both sexes) naturally exposed to enteric pathogens commonly linked to human growth stunting. Despite atrophy and abnormalities observed in the small intestine, poor growth trajectories and low serum tryptophan (an amino acid needed for protein and enzymes) levels were correlated with increased histopathology (microscopic tissue examination for disease manifestation) in the large intestine. This study provides insight into the mechanisms underlying EED and indicates that the large intestine may be an important target for therapeutic intervention. - Complex Decay Dynamics of HIV Virions, Intact and Defective Proviruses, and 2LTR Circles Following Initiation of Antiretroviral Therapy
In people living with HIV-1 who start antiretroviral therapy, virus in blood decreases rapidly to below detection, but remaining infected cells may become part of the latent reservoir. Researchers investigated viral decay dynamics and identified decay processes with pronounced differences between intact and defective proviruses. Infected cells that survive second-phase decay may down-regulate HIV-1 gene expression and enter the stable latent reservoir. This research provides insight into meaningful latent reservoir markers and mechanisms for elimination of cells with intact viral genomes. - Phagocytosis by an HIV Antibody Is Associated with Reduced Viremia Irrespective of Enhanced Complement Lysis
Researchers used the broadly neutralizing antibody (bNAb)10E8v4 targeting the HIV Env protein to examine the role of antibody-mediated effector and complement (C’) activity when 10E8v4 was administered prophylactically to rhesus monkeys challenged with simian–human immunodeficiency virus (SHIV). With sub-protective dosing, the researchers found a 78–88% reduction in post-acute viremia that was associated with 10E8v4-mediated phagocytosis. These results suggest that effector functions inherent to unmodified 10E8v4 contribute to therapeutic efficacy against SHIV, while C’ functions do not contribute to efficacy in this context. This research informs the design of bNAb modifications for improving the protective efficacy of this therapeutic approach against HIV. - Functional and Ultrastructural Analysis of Reafferent Mechanosensation in Larval Zebrafish
All animals need to differentiate between exafferent stimuli (caused by the environment) and reafferent stimuli (caused by their own movement). Researchers characterized how hair cells in zebrafish larvae discriminate between reafferent and exafferent signals. Dye labeling of the lateral line nerve and functional imaging was combined with ultra-structural electron microscopy circuit reconstruction to show that cholinergic signals originating from the hindbrain transmit efference copies and dopaminergic signals from the hypothalamus may affect threshold modulation. Findings suggest that this circuit is the core implementation of mechanosensory reafferent suppression in these young animals. - Genetic Control of the Pluripotency Epigenome Determines Differentiation Bias in Mouse Embryonic Stem Cells
Embryonic stem cells (ESCs) show an unlimited capacity for self-renewal and the ability to become any cell type in the body. Yet mechanisms for variation of ESCs from genetically diverse individuals remain largely unknown. Investigators studied regulation of cell state transitions in mouse ESCs derived from genetically diverse mouse strains and found differences in developmental potential of mouse ESCs in vitro. Recent experiments have shown that differences in cell-fate choice during development may be critical in predisposing individuals to complex diseases due to underlying differences in cell-type composition. - Blastocyst Development after Fertilization with In Vitro Spermatids Derived from Nonhuman Primate Embryonic Stem Cells
Researchers investigated whether functional spermatids (immature forms of sperm cells) can be derived in vitro from nonhuman primate pluripotent stem cells. Rhesus macaque pluripotent stem cells were differentiated into spermatogenic germ cell linages and matured in vitro to form spermatids that were capable of fertilizing oocytes (female or germ cells involved in reproduction) by injection. Successful in vitro preimplantation embryo development was observed in approximately 12% of zygotes. The data suggest potential mechanisms to address male infertility. - Natural Disaster and Immunological Aging in a Nonhuman Primate
Survivors of weather-related disasters exhibit early onset of age-related diseases. Investigators examined the impact of Hurricane Maria and its aftermath on immune cell gene expression in age-matched, cross-sectional samples from free-ranging rhesus macaques living on an isolated island. Living through an intense hurricane and its aftermath was associated with expression of key immune genes, dysregulated protein regulation networks, and greater expression of inflammatory immune cell–specific marker genes. These findings illuminate how natural disasters might become biologically embedded and contribute to earlier onset of disease and death. - New Resources for the Drosophila 4th Chromosome: FRT101F Enabled Mitotic Clones and Bloom syndrome helicase Enabled Meiotic Recombination
Seventy percent of the genes on the 4th chromosome of Drosophila melanogaster have human homologs that have a disease association. Yet, this chromosome is difficult to study because it lacks mitotic and meiotic recombination. Investigators developed technologies and stocks as a resource for the community, which enable genetic analysis of mutations on the 4th chromosome. - The Pigtail Macaque (Macaca nemestrina) Model of COVID-19 Reproduces Diverse Clinical Outcomes and Reveals New and Complex Signatures of Disease
Animal models that reproduce clinical outcomes of human COVID-19 disease are critical for understanding SARS-CoV-2 viral and immune dynamics. Investigators demonstrate that pigtail macaques recapitulate important features of COVID-19 and reveal new immune and viral dynamics of SARS-CoV-2 infection. - Precise Visuomotor Transformations Underlying Collective Behavior in Larval Zebrafish
Brain mechanisms involved in collective social behavior are mostly unknown. Investigators found how zebrafish transform visual cues into fast and accurate movement decisions when swimming in schools. For example, a fish might use the height of the fish surrounding it to estimate the distance to its neighbors. - Multiview Confocal Super-Resolution Microscopy
Researchers, including NIH scientists, report major improvements in confocal microscopy, a “workhorse” biomedical imaging technology. By harnessing new multiview light sensing optical methods in combination with deep-learning image reconstruction, researchers have improved the spatial and temporal resolutions, speed, and depth penetration of confocal microscopy and were able to acquire high-resolution images with less light. Researchers demonstrated their improved capabilities on fixed and live samples of single cells, fly wings, and mouse heart and brain tissue. - AAV Capsid Variants with Brain-Wide Transgene Expression and Decreased Liver Targeting After Intravenous Delivery in Mouse and Marmoset
Gene therapy using the shell (capsid) of adeno-associated viruses (AAVs) is increasingly being explored as a therapeutic option for debilitating disorders of the central nervous system. Investigators achieved organ-specific targeting of adeno-associated virus capsids after intravenous delivery in mice and marmosets. These results constitute an important step forward toward achieving the goal of engineered adeno-associated viruses that can be used to broadly deliver gene therapies to the central nervous system in humans. - BNT162b Vaccines Protect Rhesus Macaques from SARS-CoV-2
The preclinical development of two BNT162b vaccine candidates against SARS-CoV-2 was performed in rhesus macaques at the Southwest National Primate Research Center. - In Vitro and In Vivo Functions of SARS-CoV-2 Infection-Enhancing and Neutralizing Antibodies
Researchers isolated neutralizing antibodies against the receptor-binding domain (RBD) or N-terminal domain (NTD) of SARS-CoV-2 spike from COVID-19 patients. RBD and NTD antibodies mediated both neutralization and infection enhancement in vitro. RBD-neutralizing antibodies having cross-reactivity against coronaviruses protected against SARS-CoV-2 in vivo. - A Novel Tau-Based Rhesus Monkey Model of Alzheimer’s Pathogenesis
Researchers developed a new rhesus monkey model of Alzheimer’s disease by targeting the entorhinal cortex with an adeno-associated virus expressing a double tau mutation. - Cryopreservation Method for Drosophila melanogaster Embryos
Researchers reported a robust and efficient method for cryopreservation of Drosophila melanogaster embryos. The optimized method resulted in >10% of embryos developing into fertile adults after cryopreservation for 25 distinct strains from different sources.
Progress on Theme 2: Innovative Instruments and Equipment to Accelerate Research Discoveries
Programs and Activities Highlights
- S10 Instrumentation Fact Sheet
ORIP revised its fact sheet on the S10 Instrumentation Programs. ORIP updated previous program numbers and ensured that current funding opportunity due dates are accurate. This fact sheet is one of several fact sheets that serve as valuable resources for potential investigators to learn about ORIP resources and programs. - University of California, San Francisco, Site Visit
ORIP program staff conducted a virtual final site visit to the University of California, San Francisco (UCSF), on April 29, 2024, to conclude the 20-year oversight period of three grants. Grant C06RR014517 funded the renovation of the eighth floor of the Medical Sciences Building at the UCSF School of Pharmacy to house the new Pharmacogenetics Program. The space is occupied by 18 researchers, has resulted in several high-impact publications, and has attracted significant funding from private and public sources. Grant C06RR016267 supported the expansion and renovation of a biosafety level 3 facility at UCSF’s Institute of Neurodegenerative Diseases. The facility enabled faculty, postdoctoral researchers, and technicians at UCSF to further research on the human prions that cause Creutzfeldt-Jakob disease. Grant C06RR016490 was used to create the Program of Craniofacial and Skeletal Biology. The supported space houses the laboratories of five principal investigators, an administrative office, and core facilities for microarray analysis, which is a central resource for the Parnassus Mouse Microarray Consortium. - National IDeA Symposium of Biomedical Research Excellence
The ninth National IDeA Symposium of Biomedical Research Excellence (NISBRE), a national scientific meeting to showcase the scientific and training accomplishments of the Institutional Development Award (IDeA) Program of NIGMS, took place June 16–19, 2024, in Washington, D.C. The goal of the conference was to maximize opportunities for networking and collaborations; it was attended by undergraduate and graduate students, postdoctoral fellows, junior and senior faculty, and staff. A “Meet the Funders” session provided opportunities for the attendees to interact with NIH’s programmatic staff from different institutes and centers—including NIGMS, NIDDK, NCI, NIAID, and the Office of the Director—for program- and funding-related queries. NIGMS collaborates with the ORIP Division of Construction and Instruments (DCI) and co-funds grants supported by DCI’s S10 Shared Instrumentation Programs. An ORIP staff member was a panelist on the “Meet the Funders” session and shared information on programs managed by DCI and ORIP, including the S10 Instrumentation Programs and equipment programs. - University of Missouri Site Visit
ORIP program staff conducted a virtual final site visit to the University of Missouri on May 10, 2024, to conclude the 20-year oversight period of two grants. Grant C06RR018822 provided funds to establish the National Swine Research and Resource Center, which serves as a core facility for NIH-funded investigators and provides services in model curation, health monitoring, research, model creation, workshops, and training. Major research highlights include heart and kidney transplants. Grant C06RR017353 provided funding to establish the Center for Diabetes and Cardiovascular Health. The original grant objectives were to house and promote collaborations among NIH-funded investigators. Over the course of 20 years, investigators in the facility have been awarded about $100 million in funding by NIH and other sources, provided mentoring and training for more than 70 individuals, hosted 24 visiting scholars, and published nearly 300 peer reviewed articles. - Association of Biomolecular Resource Facilities Annual Meeting
An ORIP staff member presented on the S10 instrumentation programs at the Association of Biomolecular Resource Facilities meeting, held April 21–24, 2024, in Minneapolis, Minnesota. The presentation was attended by more than 100 people. ORIP also was represented on a panel in a breakout session. The S10 programs are based on shared use, which increases cost efficiency and benefits for thousands of investigators in hundreds of institutions nationwide. - Construction Site Visits
Between April and June 2024, ORIP’s Division of Construction and Instruments performed 13 final site visits to facilities funded by 21 construction grants. These facilities include animal housing facilities, medical imaging facilities, and centers focused on ophthalmology, immunology, infectious diseases, neuroscience, pharmaceutical, diabetes, cardiovascular, and behavioral research. Through these visits, the program staff were able to evaluate the impact, usage, and maintenance of these facilities during their 20-year (10-year for six American Recovery and Reinvestment Act grants) beneficial occupancy periods. - R24 Modern Equipment Fact Sheet
ORIP updated its fact sheet for the R24 Modern Equipment Program with up-to-date information in March 2024. ORIP updated previous program numbers and ensured that current notice of funding opportunity due dates are accurate. This fact sheet is one of several fact sheets that are valuable resources for potential investigators to learn about ORIP, ORIP-supported resources, and programs. - Morehouse School of Medicine Site Visit
ORIP program staff conducted a virtual final site visit to the Morehouse School of Medicine (MSM) on March 4, 2024, to conclude the 20-year oversight period of grants C06RR016561 and C06RR017571. These construction awards funded the renovation of animal care facility and research laboratory spaces that supported MSM’s broad research portfolio, including cancer, cardiovascular disease, neuroscience, HIV and other infectious diseases, and community and population health. These infrastructure updates have expanded the capacity of office and research spaces and increased funding from federal and non-federal programs. - Construction Site Visits
Between January and March 2024, ORIP’s Division of Construction and Instruments (DCI) performed 8 site visits to the recipient institutions of 11 construction (C06) grants. A closeout site visit to the recently completed Interdisciplinary Substance Use & Brain Injury Center at the University of New Mexico allowed DCI to learn about this construction project and assess the facility operation. The final site visits to 10 grant facilities, of which 5 were funded by the American Recovery and Reinvestment Act (ARRA) grants, allowed DCI to evaluate the impact, usage, and maintenance of these facilities during their 20-year (10-year for ARRA grants) beneficial occupancy. These facilities include animal housing facilities, medical imaging facilities, and centers focused on immunology, infectious disease, and eye research. - Medical College of Wisconsin Site Visit
ORIP program staff conducted a virtual final site visit to the Eye Institute building at the Medical College of Wisconsin (MCW) on February 21, 2024, to conclude the 20-year oversight period of grant C06RR016511. This project included the conversion of clinical and administrative space on the seventh floor to research laboratories and associated core support laboratories, as well as the renovation of 20-year-old research laboratories on the eighth floor. As a major component of MCW’s neuroscience initiative, this expansion in vision research efforts allowed the recruitment of more faculty and students and thus increased research and training capabilities. Major outcomes include more than 320 papers and chapters, 12 awarded patents, 17 pending patents, 3 licensed products, and 3 spin-out companies. - Reissue ORIP Concept Clearance: Shared Instrumentation Program
ORIP presented this concept clearance for reissue at the January 2024 NIH Council of Councils meeting. The initiative supports purchases of state-of-the-art, commercially available instruments to NIH-funded investigators. The program is based on shared use, which makes it cost efficient and beneficial to thousands of investigators in hundreds of institutions nationwide. - Oklahoma Medical Research Foundation Site Visit
ORIP program staff conducted a virtual final site visit to the Oklahoma Medical Research Foundation (ORMF) on March 18, 2024, to conclude the 20-year oversight period of grants C06RR014570, C06RR017446, and C06RR030593. These three awards provided funding to significantly renovate existing spaces and construct new spaces that have facilitated OMRF’s research on rheumatology, sarcoidosis, and autoimmune disorders like lupus and rheumatoid arthritis. ORMF reported that the funded facilities added significant funds in sponsored research and helped fund two new research centers. Additionally, OMRF scientists co-founded four companies, two of which have reached commercialization of their new technologies. - Limited Competition: Development and Renovation of Facilities for Expanding the Breeding Capacity of Specific Pathogen Free Non-Human Primates to Support HIV/AIDS–Related Research (C06 Clinical Trial Not Allowed)
Nonhuman primates (NHPs) serve as a critical animal model for biomedical research, including HIV-related research. The COVID-19 pandemic and export bans from Asian countries exacerbated the existing NHP shortage. Despite NIH efforts to improve NHP availability, limited facility resources still hamper the safe and effective production of this animal model. To meet this urgent need, ORIP, in collaboration with the Office of AIDS Research, published a new notice of funding opportunity titled Development and Renovation of Facilities for Expanding the Breeding Capacity of Specific Pathogen Free Non-Human Primates to Support HIV/AIDS–Related Research. This funding opportunity is expected to expand the support of NHP breeding resources and increase the output of NHP models substantively and cost-effectively, addressing the urgent need of NHPs for HIV research. - Site Visit to the Gladstone Institute
ORIP staff conducted a virtual site visit on November 13, 2023, to evaluate the progress of a construction award (C06RR018928) to the Gladstone Institute. This grant provided funding for the fit-out of animal facilities in the new laboratory building at the University of California, San Francisco, Mission Bay campus to support research on cardiovascular, neurological, viral, and immune diseases. The facility has supported 30 laboratories for animal studies since 2003 and enabled research funded by more than $145 million in NIH grants. Gladstone includes more than 500 researchers, including 2 Nobel laureates, and 80% of the faculty have benefitted from the animal facility. Gladstone has produced about 300 issued U.S. and foreign patents; one-third of these patents relate to animal models. - Pre-Application Webinar for Non-Human Primate Facilities (PAR-24-033) and Biomedical Research Facilities (PAR-23-306)
ORIP held a pre-application webinar in December 2023 for two notices of funding opportunities: Development and Renovation of Facilities for Expanding the Breeding Capacity of Specific Pathogen Free Non-Human Primates to Support HIV/AIDS-related Research (C06 Clinical Trial Not Allowed) and Biomedical Research Facilities (C06 Clinical Trial Not Allowed). ORIP program staff presented a program overview; explained the goals and application structures of these funding opportunities to around 130 attendees; and answered questions regarding budget, timeline, and review. Follow-up questions sent by email also were answered by ORIP program staff. - ORIP’s Construction Site Visits
Between October and December 2023, ORIP’s Division of Construction and Instruments performed site visits to eight facilities funded by construction grants. The final site visits to six of these facilities—including the animal facility at the Gladstone Institutes and the Cell Manipulation Core Facility at the Dana-Farber Cancer Institute—allowed ORIP to evaluate the impact, usage, and maintenance of these facilities during their 20-year beneficial occupancy periods. The other two were closeout site visits to two recently completed grants, which allowed ORIP to learn from these construction projects and assess the facility operation. The new Healthy Kentucky Research Building at the University of Kentucky supports research on health disparities, with an emphasis on vascular diseases. The Duke Human Vaccine Institute’s current Good Manufacturing Practice facility is at the forefront of efforts related to rapid responses and pandemic preparedness in helping the National Institute of Allergy and Infectious Diseases produce influenza and pan-coronavirus vaccine candidates. - ORIP R24 Modern Equipment Program for Innovation of Biomedical Research Operations
ORIP established the R24 Modern Equipment Program in 2022 to complement the S10 Shared Instrumentation Programs. The R24 program supports the acquisition and installation of research-enabling equipment to enhance operations in shared biomedical research facilities. To broaden the program impact, ORIP formed a collaboration with NIGMS for co-funding. This coalition resulted in 27 equipment awards in fiscal year 2023 (FY23), an increase of 6 grants from fiscal year 2022 (FY22). Awards for research capability–building in resource-limited institutions, such as the Institutional Development Award (IDeA), also increased, which resulted in 9 grants being awarded to IDeA institutions in FY23, compared to 4 awards in FY22. - ORIP S10 Shared Instrumentation Programs for Acceleration of Scientific Discoveries
ORIP’s S10 Shared Instrumentation Programs support the purchase of state-of-the-art commercially available instruments to enhance and revolutionize NIH funded research. Strategically emphasizing advanced emerging technologies, ORIP awarded 121 instruments to 74 institutions located in 29 states in fiscal year 2023. A significant number of these awards were made in collaboration with NIGMS, NIMH, NIA, OAR, and ODS, reflecting a growing recognition of the impact of the S10 programs within NIH. Among these awards, 5 were awarded to institutions from Institutional Development Award (IDeA)–eligible states, and 10 were awarded to resource-limited institutions, enhancing their research capacity and resource equity. S10 instruments are used on a shared basis, each supporting an average of 17 projects. This demonstrates the program’s operational efficiency, cost effectiveness, and broad impact on NIH research discoveries in almost all areas of science, such as cancer, general medicine, infectious diseases, neurological diseases, and aging. - ORIP C06 Construction Programs for Modernization of Biomedical Research Facilities
ORIP manages the highly competitive extramural C06 Construction Programs for the NIH Office of the Director. As a major resource to modernize the nation’s biomedical research infrastructure, ORIP’s construction programs support research capability–building projects at Institutions of Emerging Excellence (IEE) and research-intensive institutions in all geographic regions of the nation. NIH awarded 11 meritorious construction projects in fiscal year 2023, two of which are notable high-impact IEE projects. First, the Incubator and Technology Transfer Center at the University of Puerto Rico accelerates the translation of basic research concepts into job creation, commercialization, and economic growth. The second noteworthy IEE project is a chronic disease research and training facility at Stillman College, a historically Black college or university (HBCU). The new facility will promote research pertaining to stroke, heart disease, and health disparities. Additional awarded projects include pathogen-free nonhuman primate resources, biomedical imaging facilities, a cyclotron resource, a pathogen-free zebrafish facility, and a biosafety level 3 core facility. - S10 Instrumentation Programs Co-Funding
ORIP is now co-funding S10 shared instrumentation awards in collaboration with the National Institute of Mental Health. This partnership reflects ORIP’s commitment to fostering collaboration with other NIH Institutes, Centers, and Offices to promote co-funding for shared instrumentation resources. - Virtual Construction Site Visits
Between April and June 2023, ORIP’s Division of Construction and Instruments conducted six virtual site visits for ten construction grants, nine of which were near the end of their 10- or 20-year federal oversight periods. The final site visits were held to collect information on the overall impact, usage, and maintenance of the grant-supported spaces. A closeout site visit to a recently completed grant was performed to confirm the use of the facility and learn about the construction project and facility operation. All of these grant-supported spaces continue to be used for biomedical research as proposed in their original applications. - Construction and Renovation Site Visits for Awarded C06 and G20 Projects
ORIP performed more than 70 final site visits virtually in 2021, covering more than 90 construction/renovation projects that are near the end of their 10- or 20-year federal oversight periods. Information on overall impact, usage, and maintenance of grant-supported spaces is being reviewed. Most of these grant-supported spaces continue to be used for the biomedical research proposed in their original applications. - NIH Shared Instrumentation Grant Programs in 2022 NIH Annual Asian American, Native Hawaiian/Pacific Islander (AANHPI) Health Research Conference
ORIP participation at the AANHPI Health Research Conference, held May 4–5, 2022, included a presentation on the impact of and funding opportunity announcements for the NIH Shared Instrumentation Programs - Virtual Site Visits for Awarded C06 Construction Projects
Between October and December 2022, ORIP’s Division of Construction and Instruments performed four virtual site visits for seven construction projects that are near the end of their 10- or 20-year federal oversight periods. Information on overall impact, usage, and maintenance of grant-supported spaces was presented. All of these grant-supported spaces continue to be used for biomedical research as proposed in their original applications. - Funding Opportunity Announcement (FOA): PAR-22-190 (Modern Equipment for Shared-use Biomedical Research Facilities: Advancing Research-Related Operations (R24 Clinical Trials Not Allowed))
ORIP reissued the R24 FOA in July 2022 to encourage applications that seek support to purchase advanced equipment that will enhance and modernize research-supporting operations of biomedical research facilities. Laboratory research core facilities, animal research facilities, and other similar shared-use research spaces are of interest. This effort complements the existing instrument programs and broadens the support of research operation by adding modern, technologically advanced equipment to ORIP grant portfolios. - Rigor and Reproducibility of Animal Studies: Extrinsic Factors Workshop
ORIP held a three-day NIH-wide workshop in September 2022. The major goal of the workshop was to discuss the current status, needs, and strategies related to management, monitoring, and reporting of extrinsic factors to enhance the reproducibility and rigor of animal research, with a focus on ORIP-supported resources (e.g., construction, equipment, instrumentation). The Workshop was well attended with more than 600 attendees. - Virtual Construction and Renovation Site Visits for Awarded C06/G20 Projects
ORIP has held more than 44 final site visits virtually in 2022, covering more than 62 construction and renovation projects that are near the end of their 10- or 20‑year federal oversight periods. Information on overall impact, usage, and maintenance of grant-supported spaces is being reviewed. Most of these grant‑supported spaces continue to be used for the biomedical research proposed in their original applications - NIH Shared Instrumentation Grant Programs in 2022 NIH Annual Asian American, Native Hawaiian/Pacific Islander (AANHPI) Health Research Conference
ORIP participation at the AANHPI Health Research Conference, held May 4–5, 2022, included a presentation on the impact of and funding opportunity announcements for the NIH Shared Instrumentation Programs - Modern Equipment for Shared-Use Biomedical Research Facilities: Advancing Research-Related Operations
A new equipment program through ORIP's Division of Construction and Instruments, announced under PAR-21-326 (Modern Equipment for Shared-use Biomedical Research Facilities: Advancing Research-Related Operations), was launched in fiscal year 2022. Applications received for this first round were reviewed in early March. Based on the applications received, a range of equipment designed to support operational innovations in both animal facilities and core laboratories was submitted. In the applicant pool, 50% requested animal caging, 20% requested miscellaneous equipment for animal facilities, and 30% requested support equipment for core laboratories. Applications came from a total of 37 states, including 12 Institutional Development Award states. - Instrumentation Grant Programs
ORIP’s S10 Instrumentation Grant Programs support purchases of state-of-the-art commercially available instruments to enhance research of NIH-funded investigators. In fiscal year 2022, ORIP issued three funding opportunity announcements. These will be open for 3 years, with receipt dates in June each year.
- Basic Instrumentation Grant Program (BIG) – The BIG Program funds grant awards in the $25,000 to $250,000 range.
- Shared Instrumentation Grant Program (SIG) – The SIG Program funds grant awards in the $50,000 to $600,000 range.
- High-End Instrumentation Grant Program (HEI) – The HEI Program funds grant awards in the $600,001 to $2,000,000 range
- Modern Equipment for Shared-Use Biomedical Research Facilities: Advancing Research-Related Operations
ORIP’s Division of Construction and Instruments’ new equipment program was launched in fiscal year 2022 under PAR-21-326. Applications for the inaugural round were reviewed in early March 2022. Applications came from a total of 37 states, including 12 Institutional Development Award states (states that historically have had low levels of NIH funding). Applications covered a range of equipment designed to support operational innovations in both animal facilities and core labs, including animal caging, miscellaneous equipment for animal facilities, and support equipment for core laboratories. - ORIP Research Instrumentation Awards (S10)
ORIP S10 shared instrumentation programs awarded 128 scientific instruments to 84 domestic research institutions among 32 states and territories in 2021. Instrument types included biomedical imagers, cell analyzers, high-performance computers, electron microscopes, mass spectrometers, optical microscopes, magnetic resonance spectrometers, radiological/surgical instruments, and sequencers. - ORIP Support of the IDeA Program
ORIP partnered with the National Institute for General Medical Sciences to co-fund 11 S10 research instrument awards to academic and nonprofit research institutions in 8 Institutional Development Award (IDeA)–eligible states in 2021. The types of instruments awarded included cell analyzers, high-performance computers, electron microscopes, mass spectrometers, optical microscopes, and sequencers. - New Funding Opportunity for Institutions to Modernize the Functions and Operations of Existing Research Facilities
ORIP invited qualified academic or research institutions to apply for support to purchase and install advanced equipment to enhance and modernize research-supporting operations of biomedical research facilities (PAR-21-326). Core facilities, animal research facilities, and other similar shared-use facilities at academic and nonprofit research institutions will be supported by this program. - Virtual Construction and Renovation Site Visits for Awarded C06/G20 projects
ORIP performed more than 70 final site visits virtually in 2021 covering more than 90 construction/renovation projects that are near the end of 10- or 20-year federal oversight periods. Information on overall impact, usage, and maintenance of grant-supported spaces is being reviewed. Most of these grant-supported spaces continue to be used for the biomedical research proposed in their original applications.
Research Highlights from Investigators Using ORIP-Supported Instrumentation
- Generation of Locus Coeruleus Norepinephrine Neurons From Human Pluripotent Stem Cells
Researchers developed a method to generate human locus coeruleus norepinephrine (LC-NE) neurons from pluripotent stem cells, achieving 40–60% yield. These neurons display key characteristics, such as neurotransmitter release, pacemaker activity, and response to stimuli. Single-nucleus RNA sequencing confirmed their identity and differentiation pathway. Engineered with an NE sensor, these neurons reliably detect extracellular NE levels. This breakthrough offers a valuable platform for studying psychiatric and neurodegenerative diseases and advancing drug discovery. - Effects of Early Life Exposures to the Aryl Hydrocarbon Receptor Ligand TCDF on Gut Microbiota and Host Metabolic Homeostasis in C57BL/6J Mice
This study investigates how early life exposure to the pollutant tetrachlorodibenzofuran (TCDF) disrupts gut microbiota and impairs metabolic health in male mice. Findings show reduced levels of beneficial bacteria (Akkermansia muciniphila), short-chain fatty acids, and gut hormones in exposed mice, leading to long-term metabolic dysregulation. These disruptions were transferable to germ-free mice, suggesting the microbiome plays a crucial role in mediating the effects of pollutants. The research highlights the persistent health impacts of short-term pollutant exposure on the microbiome, with potential implications for obesity and metabolic diseases. - PI3Kγ Inhibition Circumvents Inflammation and Vascular Leak in SARS-Cov-2 and Other Infections
This study identifies the signaling protein PI3Kγ as a key regulator of immune cell recruitment and inflammation in severe infectious diseases, including SARS-CoV-2 and methicillin-resistant Staphylococcus aureus. PI3Kγ promotes immune cell trafficking and immune-suppressive transcription, contributing to tissue damage, fibrosis, and organ failure. Inhibiting PI3Kγ with the drug eganelisib improved survival in disease models by reducing inflammation, vascular leakage, and cytokine storms. These findings highlight PI3Kγ as a promising therapeutic target for mitigating inflammatory lung damage in severe infections. - Structural Basis of Cfr-Mediated Antimicrobial Resistance and Mechanisms to Evade It
This study presents high-resolution structures of the Cfr-modified bacterial ribosome, providing insights into antibiotic resistance mechanisms. Cfr-mediated methylation of A2503 in 23S ribosomal RNA causes an allosteric rearrangement of nucleotide A2062, reducing the effectiveness of peptidyl transferase center inhibitors. The structures also reveal how the antibiotics iboxamycin and tylosin engage with the modified ribosome. These findings advance our understanding of Cfr-mediated resistance and offer a foundation for developing strategies to overcome antibiotic resistance in Gram-positive bacteria. - APOE4 Homozygosity Represents a Distinct Genetic Form of Alzheimer's Disease
This study examined the impact of APOE4 homozygosity on Alzheimer's disease (AD) across clinical, pathological, and biomarker domains using data from the National Alzheimer's Coordinating Center and five large cohorts. Analyzing 3,297 male and female individuals pathologically and 10,039 clinically, the researchers found nearly all APOE4 homozygotes displayed AD pathology and higher biomarker levels by age 55, with almost full penetrance of abnormal amyloid levels by age 65. Symptom onset was significantly earlier, at 65.1 years, with biomarker changes resembling those in autosomal dominant AD. However, amyloid and tau positron emission tomography scans showed no differences in the dementia stage, suggesting distinct genetic underpinnings for APOE4 homozygotes in AD. - CryoEM Structures Reveal How the Bacterial Flagellum Rotates and Switches Direction
Bacterial chemotaxis relies on bidirectional flagellar rotation controlled by the flagellar motor supercomplex, including the cytoplasmic C-ring or switch. Researchers used cryogenic electron microscopy to resolve structures of the Salmonella enterica serovar typhimurium MS-ring and C-ring in both counterclockwise (4.0 Å) and clockwise poses (4.6 Å), and when bound to a regulator (5.9 Å). Conformational changes include a 180° shift in FliF/FliG domains, rotating the MotA/B binding site. The regulator specifically interacts with the clockwise conformation, suggesting a mechanism for reversing rotation and transmitting torque to the flagellum and advancing understanding of bacterial chemotaxis and motor function. - An Immunophenotype-Coupled Transcriptomic Atlas of Human Hematopoietic Progenitors
Single-cell multimodal approaches are revolutionizing the analysis of human hematopoietic progenitor cells. Cellular indexing of transcriptomes and epitopes by sequencing, which profiles both surface proteins and transcriptomes, was used on primary human bone marrow cells. Researchers optimized a panel of 132 antibodies from 266 candidates using titrations and machine learning. This multimodal analysis identified more than 80 distinct stem, progenitor, immune, stromal, and transitional cell types, each defined by unique surface markers and transcriptomes. The resulting data set offers flow cytometry solutions and reveals consistent cell surface markers across diverse donors. It also identifies normal marrow equivalents for acute myeloid leukemia stem cell populations, differing in clinical response. This work provides a valuable resource for hematopoietic progenitor research. - Uveal Melanoma Immunogenomics Predict Immunotherapy Resistance and Susceptibility
Immune checkpoint inhibition is effective for metastatic cutaneous melanoma but not for metastatic uveal melanoma (both sexes). Profiling 100 uveal melanoma metastases revealed potent tumor-specific tumor infiltrating lymphocytes (TILs), despite low mutational burden and prior immunotherapy resistance. These TILs showed low intratumoral T-cell receptor clonality. A transcriptomic biomarker was developed to identify and activate these quiescent TILs. Adoptive transfer of these selected TILs promoted tumor immunity in metastatic uveal melanoma patients, suggesting a new therapeutic approach when other immunotherapies fail. - Rare Genetic Variation in Fibronectin 1 (FN1) Protects Against APOEε4 in Alzheimer's Disease
This study explores protective genetic variants in APOEε4 carriers against Alzheimer’s disease (AD). Through whole-genome sequencing of elderly APOEε4 carriers (sex not specified) without dementia, the authors identified rare coding variants enriched in extracellular matrix (ECM)–related genes, including FN1 and COL6A2, which are expressed at the blood–brain barrier (BBB). The rs140926439 variant in FN1 was associated with reduced AD risk and delayed disease onset. Postmortem and in vivo studies confirmed increased FN1 and COL6A2 at the BBB in APOEε4 carriers with AD. Zebrafish models (both sexes) showed that FN1 loss of function reduced gliosis and enhanced microglial responses, suggesting the gene’s role in APOEε4-mediated AD pathology. Targeting such ECM proteins as FN1 could offer therapeutic avenues for AD. - Designer Peptide-DNA Cytoskeletons Regulate the Function of Synthetic Cells
Engineering artificial cells requires a dynamic cytoskeleton. Inspired by actin-binding proteins, researchers created peptide-DNA crosslinkers with varying designs. These peptide filaments form tunable bundles via DNA hybridization. In cell-sized droplets, the crosslinkers guide cytoskeletal structures to specific locations, controlling payload diffusion. Complementary DNA handles allow reversible payload recruitment. Heat triggers shape changes in droplets by adjusting DNA melting temperatures. This modular peptide-DNA design is a powerful strategy for assembling synthetic cells. - Tumour-Selective Activity of RAS-GTP Inhibition in Pancreatic Cancer
Broad-spectrum RAS inhibition that targets KRAS, HRAS, and NRAS mutations could benefit cancer patients with RAS-driven tumors. RMC-7977, a selective inhibitor for both mutant and wild-type GTP-bound RAS, was tested in various pancreatic ductal adenocarcinoma (PDAC) in vitro models, where more than 90% of cases involve KRAS mutations. The inhibitor showed significant anti-tumor activity with well-tolerated exposure levels. Tumors experienced apoptosis and sustained proliferative arrest, whereas normal tissues showed only temporary proliferation decreases without apoptosis. In a female mouse model, RMC-7977 extended survival, but Myc copy number gain led to resistance. Combining RMC 7977 with TEAD inhibition in vitro overcame this resistance, supporting the potential of RAS-GTP inhibition in PDAC therapy. - Prolonged Exposure to Lung-Derived Cytokines Is Associated With Activation of Microglia in Patients With COVID-19
Survivors of pneumonia, including SARS-CoV-2 pneumonia, face increased risks of cognitive dysfunction and dementia. Inflammation is key, with lung-derived pro-inflammatory cytokines affecting microglia, the brain cells linked to cognitive issues. A study measured cytokines during respiratory failure in male and female patients, noting higher cumulative cytokine exposure in COVID-19 cases. Microglia from COVID-19 patients showed activation signatures linked to inflammation. Corticosteroids reduced specific cytokine expressions in COVID 19. These findings highlight the role of prolonged lung inflammation in cognitive dysfunction, emphasizing the need to explore the role of microglia in pneumonia-related cognitive issues further. - A Distinct Fusobacterium nucleatum Clade Dominates the Colorectal Cancer Niche
Fusobacterium nucleatum (Fn), a microbiota that is typically found in the oral cavity in humans, has been shown to be enriched in human colorectal cancer (CRC) tissues. In this study, researchers performed a comparative genomics analysis of human CRC Fn strains. They found that CRC-associated strains belong predominantly to Fn subspecies animalis (Fna) clade C2, which shows enhanced metabolic potential and CRC colonization ability. Using female mice and human (sex not specified) CRC tissue and stool samples, the researchers confirmed Fna C2 enrichment in CRC. The authors suggest that Fna C2 could be a primary focus for future mechanistic studies and a potential pathway for therapeutic intervention in CRC. - Orthogonal Neural Encoding of Targets and Distractors Supports Multivariate Cognitive Control
In humans, complex mental activities require coordinating multiple forms of neural information processing. Recent studies show that individuals (male and female) can manage various forms of attention, but the neural control mechanisms remain unclear. In this study, researchers hypothesized that the brain monitors and prioritizes feature-specific processing independently. During functional magnetic resonance imaging, participants performed a task distinguishing target processing from distractor processing. Results showed that the dorsal anterior cingulate cortex encodes target and distractor difficulty separately, whereas the intraparietal sulcus has overlapping yet orthogonal representations of target and distractor coherence. These coherence representations, influenced by control demands, correlated with performance and frontoparietal activity, supporting top-down attention. This evidence suggests the neural geometry necessary for coordinating multivariate cognitive control. - Necroptosis Blockade Prevents Lung Injury in Severe Influenza
A new RIPK3 inhibitor, UH15-38, effectively blocks necroptosis in severe influenza A virus (IAV) infections, reducing lung inflammation and preventing mortality without hindering the immune response or viral clearance. Administered even late in infection, UH15-38 shows potential as a therapy for IAV-driven acute respiratory distress syndrome and related inflammatory conditions. This research suggests RIPK3 inhibition could offer clinical benefits for managing severe IAV infections. - Cell-Surface ZnT8 Antibody Prevents and Reverses Autoimmune Diabetes in Mice
Type 1 diabetes (T1D) involves autoimmune destruction of insulin-producing β‑cells, with ZnT8 as a key autoantigen. Monoclonal antibody mAb43 targets ZnT8, masking its antigenic sites on β-cells after insulin secretion. Administering mAb43 in non-obese diabetic female mice increased regulatory T cells in the islets, protecting against T1D onset and reversing new-onset diabetes without adverse effects. mAb43’s protective effect was reversible after stopping treatment. This study suggests mAb43 as a novel immunotherapy for preventing and reversing T1D by blocking the autoimmune attack on β-cells. - Targeting Pancreatic Cancer Metabolic Dependencies Through Glutamine Antagonism
Pancreatic ductal adenocarcinoma (PDAC) cells rely on glutamine (Gln) for proliferation and redox balance. Traditional Gln metabolism inhibitors faced resistance. Yet, treating PDAC cells with 6-diazo-5-oxo-L-norleucine (DON) induced metabolic crisis in vitro. In mice models (of unspecified sex), the pro-drug version, sirpiglenastat (DRP-104), reduced tumor growth without DON’s toxicity, enhancing survival in combination with trametinib, an extracellular signal-regulated kinase (ERK) pathway inhibitor. These findings suggest broad Gln metabolism targeting as a therapeutic strategy for PDAC, with potential enhancement through ERK pathway inhibition, offering promising avenues for treatment. - Sensory Neurons Promote Immune Homeostasis in the Lung
JAK1-dependent type 2 cytokines contribute to chronic inflammatory diseases. Inserting a human JAK1 gain-of-function variant into both male and female mice led to spontaneous atopic dermatitis–like skin disease but unexpected resistance to lung inflammation when expression was stroma-specific. JAK1 in vagal sensory neurons suppressed airway inflammation by regulating Calcb/CGRPβ expression, which in turn suppressed group 2 innate lymphoid cell function and allergic airway inflammation. These findings suggest tissue-specific roles of JAK1 in sensory neurons, offering insights for precision medicine through optimized JAK inhibitors. - Integrative Single-Cell Characterization of a Frugivorous and an Insectivorous Bat Kidney and Pancreas
This study examines cellular and molecular differences in the kidneys and pancreases of insectivorous and frugivorous bats using single-cell sequencing. Frugivorous bats show changes in kidney cell populations related to fluid and electrolyte balance, and in pancreas cell populations associated with insulin regulation. These bats exhibit molecular characteristics similar to human diabetes. Understanding these differences may offer therapeutic insights. This research sheds light on the evolution of frugivory in mammals and its implications for human health. - Serine-129 Phosphorylation of Α-Synuclein Is an Activity-Dependent Trigger for Physiologic Protein–Protein Interactions and Synaptic Function
Phosphorylation of α-synuclein at serine-129 (α-syn Ser129P) is a hallmark of synucleinopathies. Unlike wild-type α-syn, α-syn Ser129P exhibits restricted brain expression, indicating intrinsic regulation. Surprisingly, inhibiting Ser129P impairs activity-dependent synaptic attenuation by α-syn, revealing its normal function. Neuronal activity enhances Ser129P, promoting protein–protein interactions crucial for α-syn function at synapses. AlphaFold2-driven modeling and simulations suggest Ser129P induces conformational changes facilitating interactions with binding partners. This study provides insights into the role of Ser129 in synucleinopathies and potential implications for drug development. - Viral Afterlife: SARS-CoV-2 as a Reservoir of Immunomimetic Peptides That Reassemble Into Proinflammatory Supramolecular Complexes
The mechanism behind severe inflammatory responses in severe COVID-19 remains unclear. Researchers investigate the inflammatory potential of fragmented viral components from high SARS-CoV-2 loads. Machine learning analysis identifies viral peptides mimicking host antimicrobial peptides (xenoAMPs), particularly LL-37. These xenoAMPs, enriched in SARS-CoV-2, assemble viral RNA into complexes capable of multivalent binding to Toll-like receptor-3, amplifying cytokine secretion in various cell types. Administration of these complexes to mice mimics inflammatory responses seen in COVID-19 patients. Understanding this mechanism sheds light on COVID-19 pathogenesis and may inform therapeutic strategies. - OXR1 Maintains the Retromer to Delay Brain Aging Under Dietary Restriction
Dietary restriction (DR) extends life span, but its mechanism is unclear. Polymorphisms in mtd, the fly homolog of OXR1, affect life span and mtd expression under DR. Knockdown of mtd inhibits DR-induced life span extension in female flies. Mtd/OXR1 interacts with the retromer, regulating protein and lipid trafficking. Loss of mtd/OXR1 destabilizes the retromer, causing endolysosomal defects. Restoring retromer function rescues life span, neurodegeneration, and endolysosomal defects. Reduced Mtd/OXR1 is associated with aging and neurological diseases. Overexpressing Mtd/OXR1 alleviates age-related visual decline and tauopathy in flies, highlighting its role in neuronal health and longevity conservation. - A Unified Watson-Crick Geometry Drives Transcription of Six-Letter Expanded DNA Alphabets by coli RNA Polymerase
This study investigates how Escherichia coli RNA polymerase (RNAP) recognizes and processes artificially expanded genetic information system (AEGIS) nucleobases. High-resolution cryogenic electron microscopy structures of RNAP elongation complexes containing AEGIS base pairs reveal shared principles with natural base pairs. The structures show RNAPs in an active state, ready for the chemistry step, with AEGIS base pairs adopting a Watson-Crick geometry. This validates the design philosophy of AEGIS base pairs and supports the hypothesis that transcriptional mismatches occur via tautomerization. The study underscores the importance of Watson-Crick complementarity in AEGIS base pair recognition. - Long COVID-19 Manifests With T Cell Dysregulation, Inflammation and an Uncoordinated Adaptive Immune Response to SARS-CoV-2
Researchers analyzed blood samples from male and female individuals with long COVID-19 and control trajectories 8 months post-SARS-CoV-2 infection. Long COVID-19 patients displayed systemic inflammation, immune dysregulation, and altered T‑cell subset distribution. They exhibited increased frequencies of CD4+ T cells poised for tissue migration, exhausted SARS-CoV-2-specific CD8+ T cells, and disrupted coordination between T- and B-cell responses. Findings suggest improper immune crosstalk in long COVID-19, contributing to symptoms. This study sheds light on pathophysiology, potentially informing therapeutic strategies and improving patient outcomes in managing this debilitating condition. - Tumor Explants Elucidate a Cascade of Paracrine SHH, WNT, and VEGF Signals Driving Pancreatic Cancer Angiosuppression
This study presents a key mechanism that prevents pancreatic ductal adenocarcinoma (PDAC) from undergoing neoangiogenesis, which affects its development, pathophysiology, metabolism, and treatment response. Using human and murine PDAC explants (sex unknown), which effectively retain the complex cellular interactions of native tumor tissues, and single-cell regulatory network analysis, the study identified a cascade of three paracrine pathways as a key suppressor of angiogenesis in KRAS-mutant PDAC cells. This study provides an experimental paradigm for dissecting higher-order cellular interactions in tissues and has implications for PDAC treatment strategies. - Motile Living Biobots Self-Construct From Adult Human Somatic Progenitor Seed Cells
Anthrobots, spheroid-shaped biological robots derived from adult human lung cells (sex unknown), self-construct into multicellular biobots with cilia-powered locomotion. They exhibit diverse behaviors and morphologies, influenced by microenvironmental cues, without genetic editing. Anthrobots can repair scratches in human neural cell sheets, suggesting biomedical potential. This study illuminates plasticity in adult somatic cells and the potential for diverse body shapes and behaviors in living constructs. Anthrobots offer a novel platform for studying morphogenetic processes and discovering structures with biomedical relevance. - Obesity Causes Mitochondrial Fragmentation and Dysfunction in White Adipocytes due to RalA Activation
High-fat diet (HFD) induces mitochondrial fragmentation in white adipocytes via increased expression and activity of the small GTPase RalA. This fragmentation reduces oxidative capacity and promotes weight gain. Targeted deletion of RalA prevents fragmentation, enhances fatty acid oxidation, and reduces HFD-induced weight gain in mice. Mechanistically, RalA promotes mitochondrial fission by dephosphorylating Drp1. Adipose tissue expression of human Drp1 homolog, DNM1L, correlates with obesity and insulin resistance. Chronic RalA activation suppresses energy expenditure by promoting excessive mitochondrial fission, contributing to metabolic dysfunction in obesity. - Epigenetic Dysregulation From Chromosomal Transit in Micronuclei
The authors reported a mechanistic link between epigenetic alterations and chromosomal instability induced during their transit in micronuclei, both being hallmarks of advanced and metastatic cancers. It was demonstrated that the landscape of histone post-translational modifications was profoundly changed due to missegregation of mitotic chromosomes, their sequestration in micronuclei, and subsequent rupture of the micronuclear envelope. The transcriptional redistribution was attributed to micronuclei’s strong positional bias with increased promoter accessibility. The continuous formation and reincorporation of micronuclei promotes epigenetic reprogramming and heterogeneity in cancer. - Association of Age at Menopause and Hormone Therapy Use With Tau and β‑Amyloid Positron Emission Tomography
To understand the predominance (70%) of women among individuals with Alzheimer’s disease, the investigators studied regional tau and β-amyloid (Aβ) in relation to age at menopause and hormone therapy (HT) in postmenopausal women and age-matched men using positron emission tomography. The study demonstrated that females exhibited higher tau deposition compared with age-matched males, particularly in the setting of elevated Aβ; earlier age at menopause and late initiation of HT were associated with increased tau vulnerability. This study suggests women with these conditions may be at higher risk of pathological burden. The article also calls for a mechanistic understanding of menopause and dementia to help clarify the risk-benefit controversies around HT, which could be explored in future studies. This article was highlighted in various news outlets. - Broad Receptor Tropism and Immunogenicity of a Clade 3 Sarbecovirus
Investigators showed that the S glycoprotein of the clade 3 sarbecovirus PRD-0038 in the African Rhinolophus bat has broad ACE2 (angiotensin-converting enzyme 2) usage and that receptor-binding domain (RBD) mutations further expand receptor promiscuity and enable human ACE2 utilization. They generated a cryogenic electron microscopy structure of the RBD bound to ACE2, explaining receptor tropism and highlighting differences between SARS-CoV-1 and SARS-CoV-2. PRD‑0038 S vaccination elicits greater titers of antibodies cross-reacting with vaccine-mismatched clade 2 and clade 1a sarbecoviruses, compared with SARS-CoV-2. These findings underline a potential molecular pathway for zoonotic spillover of a clade 3 sarbecovirus, as well as the need to develop pan-sarbecovirus vaccines and countermeasures. - Intestinal Microbiota Controls Graft-Versus-Host Disease Independent of Donor–Host Genetic Disparity
Allogeneic hematopoietic stem cell transplantation is a curative therapy for hematopoietic malignancies and non-malignant diseases, but acute graft-versus-host disease (GVHD) remains a serious complication. Specifically, severe gut GVHD is the major cause of transplant-related mortality. Here, the authors show that genetically identical mice, sourced from different vendors, had distinct commensal bacterial compositions, which resulted in significantly discordant severity in GVHD. These studies highlight the importance of pre-transplant microbiota composition for the initiation and suppression of immune-mediated pathology in the gastrointestinal tract, demonstrating the impact of non-genetic environmental determinants on transplant outcome. - First-in-Human ImmunoPET Imaging of COVID-19 Convalescent Patients Using Dynamic Total-Body PET and a CD8-Targeted Minibody
Developing noninvasive methods for in vivo quantification of T cell distribution and kinetics is important because most T cells reside in the tissue. Investigators presented the first use of dynamic positron emission tomography (PET) and kinetic modeling for in vivo measurement of CD8+ T cell distribution in healthy individuals and COVID-19 patients. Kinetic modeling results aligned with the expected T cell trafficking effects. Tissue-to-blood ratios were consistent with model net influx rates and flow cytometry analysis. These results provide a promising platform for using dynamic PET to study the total-body immune response and memory. - Cell-Type Memory in a Single-Cell Eukaryote Requires the Continuous Presence of a Specific Transcription Regulator
Investigators studied the fundamental question and underlying mechanism of how a eukaryotic cell type can be stably maintained through many rounds of DNA replication and cell division in a fungal species where two different cell types (i.e., white and opaque) arise from the same genome. Investigators found that destruction of Wor1, the primary transcription activator of the opaque state, led opaque cells to irreversibly lose their memory and switch to the white-cell state within about 1 hour. This study demonstrates that the continuous presence of Wor1 is needed to maintain the opaque cell state. Data also suggested that a threshold concentration of Wor1 is needed to maintain the opaque state, demonstrating the importance of the transcription factor in maintaining cell-type memory. - Multimodal Single-Cell and Whole-Genome Sequencing of Small, Frozen Clinical Specimens
Single-cell RNA sequencing has led to improved understanding of tumor heterogeneity to drug response, but the broad application of those methods remains challenging due to practical requirements that are incompatible with clinical care workflow, such as the need for large and fresh tissues. The researchers demonstrated that several single-cell genomics techniques are feasible from small, frozen tissues and provide biological data outputs similar to those collected from fresh tissue, while reducing artifactual signals and compositional biases introduced by fresh-tissue processing. These results provide a new perspective for translating these methods to clinical studies. - PIKFYVE Inhibition Mitigates Disease in Models of Diverse Forms of ALS
Investigators showed that pharmacological suppression of PIKFYVE activity reduces pathology and extends survival of animal models and patient-derived motor neurons representing diverse forms of amyotrophic lateral sclerosis (ALS). Upon PIKFYVE inhibition, exocytosis is activated to transport aggregation-prone proteins out of the cells, a process that does not require stimulating macroautophagy or the ubiquitin-proteosome system. These findings suggest therapeutic potential to manage multiple forms of ALS. - Hematopoietic Stem Cells Preferentially Traffic Misfolded Proteins to Aggresomes and Depend on Aggrephagy to Maintain Protein Homeostasis
Investigators studied the relationship between the fitness of hematopoietic stem cells (HSCs) and maintenance of proteostasis. Their findings demonstrated that HSCs preferentially depend on aggrephagy, a form of autophagy, to maintain proteostasis. When aggrephagy is disabled, HSCs compensate by increasing proteasome activity, but proteostasis is ultimately disrupted as protein aggregates accumulate and HSC function is impaired. Investigators also showed that Bag3 deficiency blunts aggresome formation in HSCs, resulting in protein aggregate accumulation, myeloid-biased differentiation, and diminished self-renewal activity, thus demonstrating Bag3 as a regulator of HSC proteostasis. HSC aging is associated with loss of aggresomes and reduced autophagic flux. Protein degradation pathways are thus configured in young adult HSCs to preserve proteostasis and fitness but become dysregulated during aging. - Elevated Transferrin Receptor Impairs T Cell Metabolism and Function in Systemic Lupus Erythematosus
Systemic lupus erythematosus (SLE) is an autoimmune disease in which dysfunctional T cells exhibit abnormalities in metabolism. Investigators performed a CRISPR screen to examine mechanisms associated with the role of excess iron in dysfunctional T cells. The transferrin receptor (CD71) was identified as differentially critical for Type 1 T helper cells and inhibitory for induced regulatory T cells. Activated T cells induced CD71 and iron uptake, which was exaggerated in SLE-prone T cells. Disease severity correlated with CD71 expression in cells from male and female patients with SLE, and blocking CD71 in vitro enhanced interleukin 10 secretion. These findings suggest that T cell iron uptake via CD71 contributes to T cell dysfunction and can be targeted to limit SLE-associated pathology. - Cannabinoid Receptor 1 Antagonist Genistein Attenuates Marijuana-Induced Vascular Inflammation
Marijuana use is increasing and is associated with increased risk of cardiovascular disease (CVD); however, the link between marijuana and CVD remains largely unknown. Investigators demonstrated that a psychoactive component of marijuana, Δ9-tetrahydrocannabinol (Δ9‑THC), activates cannabinoid receptor 1 (CB1), causing vascular inflammation, oxidative stress, endothelial dysfunction, and atherosclerosis. The in silico virtual screening study suggested that genistein, a soybean isoflavone, would be a putative CB1 antagonist. Their validation study showed that in male mice, genistein blocked Δ9-THC-induced endothelial dysfunction in wire myograph, reduced atherosclerotic plaque, and minimally penetrated the central nervous system. This study revealed for the first time that genistein is a CB1 antagonist that attenuates Δ9-THC-induced atherosclerosis while preserving clinically useful effects. - Characterizing a Photoacoustic and Fluorescence Imaging Platform for Preclinical Murine Longitudinal Studies
Preclinical studies using animal models require medical imaging technology with sufficient resolution and sensitivity for anatomical, functional, and molecular assessments. Photoacoustic (PA) tomography provides high resolution and specificity, and fluorescence (FL) molecular tomography provides high sensitivity; the combination of these imaging modalities capitalizes on their strengths and mitigates disadvantages. In this publication, the authors describe TriTom, a preclinical imaging system that integrates PA and FL. They characterized the PA spatial resolution, PA sensitivity, PA spectral accuracy, optical spatial resolution, and FL sensitivity of the platform and demonstrated anatomical imaging in mice. This report demonstrates TriTom’s suitability for biomedical imaging applications. - Gigapixel Imaging with a Novel Multi-Camera Array Microscope
Complex biological systems are organized across many spatial scales, and maintaining field of view and resolution has been a longstanding challenge. The investigators constructed a scalable multi-camera array microscope (MCAM) that allows high-resolution, large-field-of-view recording from multiple spatial scales simultaneously. The MCAM enabled the team to observe the behavior and fine anatomical features of numerous freely moving model organisms on multiple spatial scales. This work provides a scalable platform that overcomes current imaging bottlenecks and can allow researchers to observe organisms across microscopic and macroscopic scales. - Recreating the Heart’s Helical Structure–Function Relationship with Focused Rotary Jet Spinning
The investigators developed a tissue engineering approach that enables rapid deposition of cardiomyocyte microfibers with programmable alignments in 3D geometries. Using this focused rotary jet spinning (FRJS) method, they reproduced tissue scaffolds with contractile cells’ helical alignments, resembling complex structures of the musculature and properties of a natural heart. This work represents an important advance toward biofabrication of tissue models for healthy and diseased hearts by manipulating orientation of specific fibers. With the technological advancement over other competing methods, FRJS might provide a pathway for fabricating other tissues and organs with diverse cell populations. - The Ras GTPase–Activating–Like Protein IQGAP1 Bridges Gasdermin D to the ESCRT System to Promote IL‐1β Release via Exosomes
The investigators identified IQGAP1, a scaffold protein, as a gasdermin D (GSDMD)–interacting protein through a nonbiased proteomic analysis. Functional investigation indicated that the interaction is required for lipopolysaccharide (LPS)- and ATP-induced exosome release. Further analysis revealed that IQGAP1 serves as an adaptor that bridges GSDMD and the associated IL‐1β complex to Tsg101 and enables the packaging of GSDMD and IL‐1β into exosomes. This process is dependent on an LPS‐induced increase in GTP‐bound CDC42, a small GTPase known to activate IQGAP1. This study reveals IQGAP1 as a link between inflammasome activation and exosomal release of IL‐1β. - A Deep Learning Platform to Assess Drug Proarrhythmia Risk
Investigators trained a convolutional neural network (CNN) classifier to learn and identify features of in vitro action potential recordings of human induced pluripotent stem cell–derived cardiomyocytes (hiPSC-CMs) that are associated with lethal Torsade de Pointes arrhythmia. The CNN classifier accurately predicted the risk of drug-induced arrhythmia. The risk profiles of the test drugs were similar across hiPSC-CMs derived from different healthy donors. In addition, pathogenic mutations that cause arrhythmogenic cardiomyopathies in patients significantly increased the proarrhythmic propensity to certain intermediate and high-risk drugs in the hiPSC-CMs. These data indicate that deep learning can identify in vitro arrhythmic features that correlate with clinical arrhythmia and discern the influence of patient genetics on the risk of drug-induced arrhythmia. - Obesity Alters Pathology and Treatment Response in Inflammatory Disease
Obesity and metabolic disease have been shown to affect immunotherapeutic outcomes. By studying classical type 2 T helper cells (TH2) in lean and obese male mouse models for atopic dermatitis, the investigators found that the biologic therapies protected lean mice but exacerbated disease in obese mice. RNA sequencing and genome analyses revealed decreased activity of nuclear receptor PPARγ (peroxisome proliferator–activated receptor-γ) in TH2 cells in obese mice, compared to lean mice, indicating that PPARγ is required to prevent aberrant non-TH2 inflammation. Understanding the effects of obesity on immunological disease could inform a potential precision medicine approach to target the immune dysregulation caused by obesity. - Using the Autofluorescence Finder on the Sony ID7000TM Spectral Cell Analyzer to Identify and Unmix Multiple Highly Autofluorescent Murine Lung Populations
The investigators explored a new imaging approach to detect faint fluorescent signals that are masked in the background of cell types that emit high-intensity autofluorescence (AF) signals in a flow cytometry panel. Using a novel AF finder tool on the Sony ID7000™ spectral cell analyzer, the investigators studied multiple AF subsets in complex and heterogeneous murine lung single-cell suspensions. In each reiteration, the AF finder tool screens an entire single-cell population for highly autofluorescent subsets. Each identified subset’s distinct spectra are characterized and subtracted from the fluorescence parameters. Repeated subtraction of these distinct AF spectra resulted in signals of the markers of interest with a resolution increase by several orders of magnitude in some fluorescent channels. The major immune and lung tissue resident cells in a murine model for asthma were easily identified in a multicolor panel using AF subtraction. The findings demonstrate for the first time the practicality of the AF finder tool—particularly when analyzing samples with multiple AF populations of varying intensities—for reducing fluorescence background and increasing signal resolution in spectral flow cytometry. - Distinct Metabolic States Guide Maturation of Inflammatory and Tolerogenic Dendritic Cells
The investigators mapped single-cell metabolic states and immune profiles of inflammatory and tolerogenic monocytic dendritic cells using recently developed multiparametric approaches. Activation scores revealed simultaneous engagement of multiple metabolic pathways in distinct monocytic dendritic cell differentiation stages (e.g., rapid reprogramming of glycolytic monocytes and transient co-activation of mitochondrial pathways followed by maturation of dendritic cells). This single-cell data set provides important insights into metabolic pathways that affect the immune profiles of human dendritic cells. - Chronic TREM2 Activation Exacerbates Aβ-Associated Tau Seeding and Spreading
Researchers used a mouse amyloidosis model in which AD-associated tau protein is injected into the brain to induce amyloid β (Aβ)–dependent tau seeding/spreading. They found that in male and female animals, chronic administration of an activating TREM2 (triggering receptor expressed on myeloid cells 2) antibody increases microglial activation of dystrophic surrounding neuritic plaques (NPs), a subtype of Aβ plaques, but increases NP-tau pathology and neuritic dystrophy without altering Aβ plaque burden. These data suggest that sustained microglial activation through TREM2 that does not result in strong myeloid removal might exacerbate Aβ-induced tau pathology, which could have important clinical implications. - Intrahost Evolution and Forward Transmission of a Novel SARS-CoV-2 Omicron BA.1 Subvariant
Investigators studied persistent SARS-CoV-2 infections in immune-compromised patients to test the hypothesis that intrahost viral evolution in SARS-CoV-2 variants, such as the Omicron variant, can be attributed to suboptimal immune responses. In a male immune-compromised individual with prolonged SARS-CoV-2 infection (during a 12-week period), investigators detected the emergence of eight additional amino acid substitutions within the spike protein of the Omicron BA.1 sub-lineage. They observed five subsequent cases in individuals of both sexes (three of whom were in the same health care system) bearing this unique combination of mutations, indicative of forward transmission. The study confirmed intrahost evolution of SARS-CoV-2 occurring during persistent infection in immunocompromised individuals; this evolution can drive the emergence and spread of new subvariants with extensive mutations in key antigenic regions. This study also highlights the urgent need to develop novel therapeutic strategies to limit duration of infection and viral spread in vulnerable patients. - Wastewater Sequencing Reveals Early Cryptic SARS-CoV-2 Variant Transmission
The investigators explored the use of SARS-CoV-2 RNA concentration in wastewater as a practical approach to estimating community prevalence of COVID-19, detecting emerging variants, and tracking regional infection dynamics. Two obstacles must be overcome to leverage the wastewater-based genomic surveillance: low-quality sequence data and inability to estimate relative lineage abundance in mixed samples. The investigators developed and deployed improved virus concentration protocols and deconvolution software to fully resolve multiple virus strains from wastewater. Results indicate that emerging variants of concern were detected up to 14 days earlier in wastewater samples than via genomic clinical surveillance, and multiple instances of virus spread that were not captured by clinical genomic surveillance were identified by the wastewater-based genomic surveillance. The study provides a scalable solution for wastewater genomic surveillance that allows early detection of SARS-CoV-2 variants and identification of cryptic transmission. The work suggests a critical, needed methodology for early detection of emerging variants and swift public health interventions. - Using the Autofluorescence Finder on the Sony ID7000TM Spectral Cell Analyzer to Identify and Unmix Multiple Highly Autofluorescent Murine Lung Populations
The investigators explored a new imaging approach to detect faint fluorescent signals that are masked in the background of cell types that emit high-intensity autofluorescence (AF) signals in a flow cytometry panel. Using a novel AF finder tool on the Sony ID7000™ spectral cell analyzer, the investigators studied multiple AF subsets in complex and heterogeneous murine lung single-cell suspensions. In each reiteration, the AF finder tool screens an entire single-cell population for highly autofluorescent subsets. Each identified subset’s distinct spectra are characterized and subtracted from the fluorescence parameters. Repeated subtraction of these distinct AF spectra resulted in signals of the markers of interest with a resolution increase by several orders of magnitude in some fluorescent channels. The major immune and lung tissue resident cells in a murine model for asthma were easily identified in a multicolor panel using AF subtraction. The findings demonstrate for the first time the practicality of the AF finder tool—particularly when analyzing samples with multiple AF populations of varying intensities—for reducing fluorescence background and increasing signal resolution in spectral flow cytometry. - Understanding the Computational Demands Underlying Visual Reasoning
Comprehension of the complex visual relations between objects is necessary for visual reasoning. In this study, investigators sought to better understand the computational demands involved in this process. They systematically assessed the ability of modern deep convolutional neural networks (CNNs) to learn to solve the synthetic visual reasoning test (SVRT) challenge. They also extended the CNNs with spatial- and feature-based attention mechanisms and evaluated the ability of the networks to learn to solve the SVRT challenge. They found that the corresponding improvements on individual tasks partially explained their novel taxonomy. This work yields testable neuroscience predictions on differential needs, depending on the type of visual reasoning problem. - High-Fat Diet Causes Inferior Vertebral Structure and Function without Disc Degeneration in RAGE-KO Mice
Back pain and spinal pathologies are associated with obesity in juveniles and adults, but the causal relationships associated with this effect are largely unknown. Using a RAGE-KO (receptor for advanced glycation end-products knockout) mouse model of both sexes, researchers examined the association between high-fat diet and structural and functional changes to vertebrae and intervertebral discs (IVDs). The high-fat diet resulted in inferior vertebral structure and function with some sex differences, no IVD degeneration, and few modifying effects of RAGE‑KO. These findings suggest that diet can affect spinal structures and might increase the risk for spinal injury and degeneration with aging and additional stressors. - A Vaccine Targeting Resistant Tumours by Dual T Cell Plus NK Cell Attack
Most cancer vaccines target peptide antigens, and tumours frequently escape T cell–mediated immunity through mechanisms that interfere with peptide presentation. The investigators developed a new cancer vaccine that maintains efficacy against major histocompatibility complex class I–deficient tumours that are resistant to cytotoxic T cells through the coordinated action of NK (natural killer) cells and CD4+ T cells. The vaccine also inhibits the outgrowth of metastases following surgical removal of primary, highly metastatic tumours. These findings suggest that the new design enables protective immunity even against tumours with common escape mutations. - Matrisome Changes in Parkinson’s Disease
Changes in the extracellular matrix proteins—known collectively as the matrisome—are associated with Parkinson’s disease (PD). Investigators analyzed proteomics data from the prefrontal cortex in patients with PD. They also performed histological analysis. Enrichment of type 1 collagen was found to be associated with PD in both experimental approaches. Additional findings suggest that protein-linked glycans and angiogenesis might play a role in PD. Histological studies can provide a complementary technique for future large-cohort studies of the matrisome. Additionally, this approach could be applied for selecting regions of interest for proteomic analysis. - Cryo-EM Structure of a Kinetically Trapped Dodecameric Portal Protein from the Pseudomonas-Phage PaP3
Pseudomonas-phages have been recognized for their biomedical relevance, but few studies in this area have been performed to date. Investigators used cryogenic electron microscopy (cryo-EM) single-particle analysis to determine the near-anatomic structure of the portal protein of the Pseudomonas-phage PaP3. This structural analysis revealed that the portal exists as a dodecamer with helical symmetry, rather than rotational. This work established the structural repertoire of Pseudomonas-phage portals and could offer insight into the conformational dynamics of the portal barrel. Additionally, the investigators demonstrated the successful use of cryo-EM in exploring such macromolecular assemblies. - Toxic SOD1 Trimers Are Off-Pathway in the Formation of Amyloid-Like Fibrils in ALS
The aggregation of proteins into large insoluble species is a hallmark of many neurodegenerative diseases, but recent evidence suggests that smaller soluble aggregates—such as nonnative superoxide dismutase (SOD1) trimeric oligomers—are responsible for neuronal death. Both factors must be considered in the development of therapeutics for neurodegenerative diseases, such as amyotrophic lateral sclerosis (ALS). Investigators stabilized the trimeric form of SOD1, which decreased the formation of larger aggregates while increasing toxicity to cells. They showed that that trimeric SOD1 is off-pathway in the formation of protective SOD1 inclusion bodies. These results provide insight into the design of therapeutic interventions for ALS. - Basis of Narrow-Spectrum Activity of Fidaxomicin on Clostridioides difficile
Fidaxomicin, an RNA polymerase inhibitor, is used to treat Clostridioides difficile infection. The molecular basis of the pathogen’s narrow-spectrum activity in the gut microbiome, however, is unknown. Investigators used cryogenic electron microscopy to determine the structure of C. difficile RNA polymerase in complex with fidaxomicin. They identified a crucial fidaxomicin-binding determinant of the RNA polymerase that is absent in most gut microbiota. Structural, biochemical, genetic, and bioinformatic analyses were used to identify the sensitizing element for fidaxomicin’s narrow-spectrum activity. These findings can be applied toward a targeted C. difficile drug design. - A Naturally Inspired Antibiotic to Target Multidrug-Resistant Pathogens
Antibiotic resistance has contributed to an increasing number of deaths associated with gram-negative bacterial infections. Colistin, a naturally derived antibiotic, is used as a last-resort treatment; however, the recent spread of mobilized colistin-resistance gene (mcr-1) might lessen colistin’s usefulness. Researchers proposed that a solution to mcr-1-mediated resistance might have evolved among naturally occurring colistin congeners. They conducted a bioinformatic analysis and identified a biosynthetic gene cluster that is predicted to encode a structurally divergent colistin congener. The researchers synthesized macolacin using this structure and provided evidence for its utility in a mouse neutropenic infection model. -
Omicron Spike Function and Neutralizing Activity Elicited by a Comprehensive Panel of Vaccines
The SARS-CoV-2 Omicron variant comprises several sublineages, including BA.1, BA.2, BA.3, BA.4, and BA.5. Several of these sublineages differ in the sequence of the spike glycoprotein, which interacts with the receptor angiotensin-converting enzyme 2 (ACE2). The investigators aimed to understand how the different spike mutations in the Omicron variant sublineages affect host receptor engagement and membrane fusion. They showed that the mutations affect ACE2 binding, fusogenicity, and plasma-neutralizing activity in response to infection or clinical vaccines. These data indicate the effectiveness of vaccine boosters in conferring protection against Omicron-induced severe disease.
- A Molecular Analysis of Memory B Cell and Antibody Responses Against Plasmodium falciparum Merozoite Surface Protein 1 in Children and Adults from Uganda
Novel interventions for malaria, including an efficacious vaccine, are critically needed to reduce the incidence of disease. In this study, researchers sought to understand how memory B cell and antibody responses develop over the course of lifelong Plasmodium falciparum exposure. They demonstrated the evolution of full-length P. falciparum merozoite surface protein 1 (PfMSP1)–specific humoral immune response with cumulative pathogen exposure. They reported a shift from IgM+ to IgG+ B cell memory, diversification of B cells from germline, and stronger recognition of PfMSP1 variants by the plasma IgG repertoire. These results help elucidate the development of B-cell responses to P. falciparum infection. - Dual-Function Antibody Conjugates for Multimodal Imaging and Photoimmunotherapy of Cancer Cells
Precision imaging is an important tool in cancer diagnostics and guiding therapies. The investigators proposed a dual-function antibody conjugate (DFAC) comprising an FDA-approved photosensitizer and a naphthalocyanine-based photoacoustic dye for multimodal infrared imaging and demonstrated the utilization of DFACs for multimodal imaging and photodynamic treatment of squamous cell carcinoma cell line. The proposed DFACs—which were generated by complementing fluorescence imaging with photoacoustic imaging—have potential use in precision imaging applications for preclinical and clinical translation, such as guiding tumor resection surgeries and photodynamic treatment of residual microscopic disease, thereby minimizing local recurrence - Structure-Based Design and Antigenic Validation of Respiratory Syncytial Virus G Immunogens
Respiratory syncytial virus (RSV) is a leading cause of severe lower respiratory tract disease of children, the elderly, and immunocompromised individuals. The investigators demonstrated the use of an RSV glycoprotein (G) variant, S177Q protein, to develop RSV vaccines. The RSV G S177Q protein retains high-affinity binding to protective human and mouse monoclonal antibodies and has equal reactivity as wild-type RSV G protein to human reference immunoglobulin to RSV. Additionally, they determined the high-resolution crystal structure of RSV G S177Q protein in complex with the anti-RSV G antibody 3G12, further validating its antigenic structure. These studies suggest that an engineered RSV G protein is an immunogen candidate for developing RSV vaccines - Proteomic Analysis of Microtubule Inner Proteins (MIPs) in Rib72 Null Tetrahymena Cells Reveals Functional MIPs
Microtubules are dynamic protein structures within cells that perform diverse essential functions, including providing the force required to bend cilia (hairlike structures on the outside of cells) to allow cells to move through fluid. Researchers studied the structure and function of microtubule inner protein Fap115 in the motile cilia to better understand ciliary structure and function. Using mass spectrometry, proteomic analysis, and cryo-electron tomography, the region of microtubules where Fap115 proteins localize was identified. Using a genetic knockout of Fap115 in Tetrahymena thermophila, a unicellular eukaryote, researchers revealed that loss of Fap115 function resulted in aberrant ciliary motion and disrupted cell swimming. - Mechanism of Integrin Activation by Talin and Its Cooperation with Kindlin
Talin is a cytoskeletal protein with many cellular functions. However, how talin head terminal (talin-H) operates in the context of full-length talin and its surroundings remains unknown. Investigators showed that while being capable of inducing integrin activation, talin-H alone exhibits unexpectedly low potency versus a constitutively activated full-length talin. They found that the large C‑terminal rod domain of talin (talin-R), which otherwise masks the integrin binding site on talin-H in inactive talin, dramatically enhances the talin-H potency by dimerizing activated talin and bridging it to the integrin co-activator kindlin-2. These data provide crucial insight into the mechanism of talin and its cooperation with kindlin to promote potent integrin activation, cell adhesion, and signaling - Integrated Analysis of Plasma and Single Immune Cells Uncovers Metabolic Changes in Individuals with COVID-19
A better understanding of the metabolic alterations in immune cells during infection by SARS-CoV-2 infection may elucidate the wide diversity of clinical symptoms experienced by individuals with COVID-19. Investigators reported metabolic changes associated with the peripheral immune response of individuals with COVID-19. The results revealed a robust interplay between plasma metabolites and cell type–specific metabolic reprogramming networks that were associated with disease severity and could predict survival. - IMPDH1 Retinal Variants Control Filament Architecture to Tune Allosteric Regulation
Purines are essential components of RNA and DNA and play roles in metabolism. Investigators studied a regulatory factor for making purines: inosine-5′-monophosphate dehydrogenase 1 (IMPDH1), which plays a unique role in retinal metabolism. Mutation of IMPDH1 causes human blindness. Investigators showed that canonical human IMPDH1 assembles into filaments with different structures depending on its activity state. These results provide insight into regulatory mechanisms for IMPDH1. - Immune Correlates Analysis of the mRNA-1273 COVID-19 Vaccine Efficacy Clinical Trial
Investigators determined that antibodies are the correlate of protection in vaccinated individuals enrolled in the Moderna coronavirus efficacy (COVE) Phase 3 clinical trial. Vaccine recipients were assessed for neutralizing and binding antibodies as correlates of risk for COVID-19 disease and as correlates of protection. All markers were inversely associated with COVID-19 risk and directly associated with vaccine efficacy. These results help define immune marker correlates of protection and may guide approval decisions for messenger RNA (mRNA) and other COVID-19 vaccines. - Selective G Protein Signaling Driven by Substance P–Neurokinin Receptor Dynamics
Investigators determined the cryogenic-electron microscopy structures of active neurokinin-1 receptor, a G protein–coupled receptor, bound to neuropeptide substance P (SP) or the G protein q-biased peptide SP6–11. This data unveils the molecular mechanism of how two stimuli (SP and neurokinin A) yield distinct G protein signaling at the same G protein–coupled receptor. - Spatial Transcriptomics Using Combinatorial Fluorescence Spectral and Lifetime Encoding, Imaging, and Analysis
Investigators introduced a spatialomics technology—termed Multi Omic Single-scan Assay with Integrated Combinatorial Analysis (MOSAICA)—that integrated in situ labeling of mRNA and protein markers with combinatorial fluorescence spectral and lifetime encoded probes, spectral and time-resolved fluorescence imaging, and machine learning–based decoding. Investigators demonstrated MOSAICA’s multiplexing scalability, applied this technology for multiplexed analysis, and showed simultaneous co-detection of protein and mRNA in multiple cell and tissue types. - Severely Ill COVID-19 Patients Display Impaired Exhaustion Features in SARS-CoV-2–Reactive CD8+ T Cells
Investigators studied single-cell transcriptomes of >80,000 virus-reactive CD8+ T cells from COVID-19 patients using a modified antigen-reactive T cell enrichment (ARTE) assay and found substantial diversity in the nature of CD8+ T cell responses to SARS-CoV-2 infection among patients. - Imbalance of Regulatory and Cytotoxic SARS-CoV-2–Reactive CD4+ T Cells in COVID-19
Researchers reported a study of CD4+ T cells in the immune responses of hospitalized and non-hospitalized patients with SARS-CoV-2 infection and identified distinct gene expression patterns of SARS-CoV-2–reactive CD4+ T cells associated with disease severity. - A Noncoding RNA Modulator Potentiates Phenylalanine Metabolism in Mice
Investigators demonstrated that the mouse long noncoding RNA (lncRNA) Pair and human lncRNA HULC associate with phenylalanine hydroxylase and can be utilized to develop mouse and human induced pluripotent stem cell (iPSC) models of phenylketonuria for preclinical research. - An NR2F1-Specific Agonist Suppresses Metastasis by Inducing Cancer Cell Dormancy
Investigators discovered an agonist of the nuclear receptor NR2F1 that activates dormancy programs in malignant cells. This dormancy program arrested the growth of multiple human cell lines and patient-derived organoids and inhibited lung metastases of head and neck squamous cell carcinomas in mice.
Progress on Theme 3: Specialized Research Training in Animal Models and Related Resources
Programs and Activities Highlights
- Opportunities for Veterinary Scientists at NIH
ORIP staff participated in a virtual discussion with veterinary students on opportunities for veterinary scientists at NIH, which was hosted through the University of Pennsylvania School of Veterinary Medicine on June 24, 2024. - Notice of Funding Opportunity Reissue (PA-24-176) and ORIP Information Update for PA-24-176
The purpose of the NIH Mentored Research Scientist Development Award (K01) is to provide support and protected time (3–5 years) for an intensive, supervised career development experience in the biomedical, behavioral, or clinical sciences leading to research independence. When NIH reissued the notice of funding opportunity, ORIP added specific information to the funding opportunity PA-24-176 in April 2024. For investigator-initiated applications submitted in response to the parent funding opportunity announcement—Mentored Research Scientist Development Award (Parent K01 – Independent Clinical Trial Not Allowed)—ORIP support is limited to graduate veterinarians (i.e., D.V.M.s, V.M.D.s, and equivalents) only. The K01 allows 4 years of protected time to develop an awardee’s research career. Once a K01 has been obtained, the awardee is permitted, and actively encouraged, to apply for additional research grants anytime during the tenure of their K award. - T32 and T35 Directors Consortium
Two ORIP program staff members attended the T32/T35 Directors Consortium Meeting on March 11, 2024. ORIP participated in a panel discussion on the topic of leveraging the T35 experiences to create a pipeline for qualified T32 trainees. ORIP’s T32 Postdoctoral Program for Veterinarians trains highly qualified veterinarians for research careers in biomedical areas related to comparative medicine. - Special Emphasis Research Career Award in Comparative or Translational Medicine
These guidelines summarize policies governing the Special Emphasis Research Career Award (SERCA) in Comparative or Translational Medicine, which is offered by ORIP’s Division of Comparative Medicine. Potential applicants should also refer to these guidelines for the Mentored Research Scientist Development Award (Parent K01), PA-20-190. Differences between SERCA and the Parent K01 are described in these guidelines. This award is intended solely to provide support and protected time to graduate veterinarians for an intensive, supervised career development experience in the biomedical sciences leading to research independence. - T32/T35 Directors Consortium Meeting
An ORIP program staff member attended the T32/T35 Directors Consortium Meeting on December 11, 2023. ORIP participated in a panel discussion focused on leveraging the T35 experiences to create a pipeline for qualified T32 trainees. - Veterinary Scholars Symposium 2023
A satellite meeting with T32 and T35 training program directors was held on August 4, 2023, during the Veterinary Scholars Symposium. Participants discussed challenges in recruiting and retaining veterinary students and veterinarians into research, as well as continuation of the previous R13 conference grant and development of a new R13 grant. Participants asked questions about providing support to investigators or mentors, extending support for veterinarians past 36 months, and providing support for combined residencies (clinical and research). The participants suggested that grants management representatives attend the next T32 Directors Consortium to present on allowable costs. Additionally, program reviews of the NIH Grants Policy Statement were suggested. - T32/T35 Directors Consortium Meeting
ORIP program staff members attended the T32/T35 Directors Consortium Meeting on September 18, 2023, at the University of Minnesota. Grants management issues were addressed, such as pre-award costs and stipends. Details of leveraging the T35 as a pipeline for the T32 were noted. - Host Genetic Variation Impacts SARS-CoV-2 Vaccination Response in the Diversity Outbred Mouse Population
The COVID-19 pandemic led to the rapid and worldwide development of highly effective vaccines against SARS-CoV-2. Although host genetic factors are known to affect vaccine efficacy for such respiratory pathogens as influenza and tuberculosis, the impact of host genetic variation on vaccine efficacy against COVID-19 is not well understood. Investigators used the diversity outbred mouse model (males) to study the effects of genetic variation on vaccine efficiency. Data indicate that variations in vaccine response in mice are heritable, similar to that in human populations. - Newly Identified Roles for PIEZO1 Mechanosensor in Controlling Normal Megakaryocyte Development and in Primary Myelofibrosis
Mechanisms through which mature megakaryocytes (Mks) and their progenitors sense the bone marrow extracellular matrix to promote lineage differentiation are only partially understood. The authors report that PIEZO1, a mechanosensitive cation channel, is expressed in mouse and human Mks, and activation of PIEZO1 increased the number of immature Mks in mice. Piezo1/2 knockout mice of both sexes show an increase in Mk size and platelet count, both at basal state and upon marrow regeneration. Together, these data suggest that PIEZO1 places a brake on Mk maturation and platelet formation in physiology, and its upregulation might contribute to aggravating disease. - The Gene Expression Profile and Cell of Origin of Canine Peripheral T-Cell Lymphoma
Peripheral T-cell lymphoma (PTCL) refers to a heterogenous group of T-cell neoplasms with poor treatment responses and survival times. Canine PTCL clinically and immunophenotypically resembles the most common human subtype, PTCL-NOS (PTCL-not otherwise specified) and is a naturally occurring model for human PTCL. Gene expression profiling in human PTCL-NOS has helped characterize this ambiguous diagnosis into distinct subtypes, but similar gene expression profiling in canine PTCL is lacking. Canine CD4+ PTCL most closely resembles the GATA3-PTCL subtype of PTCL-NOS and may originate from an earlier stage of T-cell development than the more conventionally posited mature T-helper cell origin. - National Veterinary Scholars Symposium
ORIP held a Training Directors’ meeting at the National Veterinary Scholars Symposium, which was held in San Juan, Puerto Rico, in August 2023. This meeting highlights the essential role of scientific research in veterinary medicine and provides veterinary medical students who have conducted original research an opportunity to present their research. - ORIP Concept Clearance: HIV/AIDS Scholars Programs (K01, R21, R13) Reissue
ORIP presented this concept clearance for reissue at the January 19–20, 2023 NIH Council of Councils meeting. The initiative will facilitate development of effective HIV/AIDS treatment and prevention by increasing the workforce that can employ nonhuman primates as preclinical models for HIV/AIDS research. - T32 and T35 Joint Director’s Consortium Meeting
ORIP provided NIH updates at the T32 and T35 Joint Consortium Meeting on March 20, 2023. Updates included information on various notices related to training (NOT-OD-22-055, NOT-OD-23-076, NOT-OD-23-084); the Advisory Committee to the Director Working Group on Re-envisioning NIH-Supported Postdoctoral Training and associated requests for information and listening sessions; an Open Mike Extramural Nexus article, “Number of Postdoctoral Researchers Supported by NIH Grant Awards FY 2017–FY 2022”; and review of prior approval actions. - Funding Opportunity Announcement: HIV/AIDS Scholars Using Nonhuman Primate (NHP) Models Program (K01 Independent Clinical Trial Not Allowed)
ORIP is partnering with the National Institute of Allergy and Infectious Diseases and National Institute of Mental Health to provide salary and research support for early-stage investigators who are working in the field of HIV/AIDS translational studies, using NHPs as preclinical models. - T32 Training Directors Consortium Meeting
The T32 Training Directors Consortium Meeting was held virtually in August 2022. ORIP program staff attended the meeting and participated in discussions of items of concern. - Webinar on Opportunities at NIH for Veterinarian-Scientists
ORIP was invited to speak on opportunities at NIH for veterinarian-scientists during a webinar that was held on June 1, 2022. The audience was composed of veterinary students who were engaged in summer research at Mississippi State University College of Veterinary Medicine and Tuskegee University College of Veterinary Medicine. Associated Deans of Research from other veterinary schools in the region—Louisiana State University School of Veterinary Medicine, Auburn University College of Veterinary Medicine, and The University of Tennessee College of Veterinary Medicine—also were invited - NIH Extramural Loan Repayment Program Webinar
ORIP participated in the NIH Extramural Loan Repayment Program Webinar, which was presented to an audience largely composed of veterinary students engaged in summer research from Michigan State University College of Veterinary Medicine and Tuskegee University College of Veterinary Medicine. Associate deans of research from other veterinary schools in the region (e.g., Louisiana State University, Auburn University, The University of Tennessee) also were invited to attend. - Research Enhancement Award Program (REAP) for Health Professional Schools and Graduate Schools
ORIP participates in the REAP for Health Professional Schools and Graduate Schools initiative to support small-scale research grants at institutions that do not receive substantial funding from the NIH. The program provides biomedical research experiences primarily for health professional, undergraduate and graduate students and enhances the research environment at applicant institutions. ORIP emphasis is given to colleges of veterinary medicine applications proposing comparative studies of a wide range of biological models to improve their value in translational research. - Research Supplements to Promote Diversity in Health-Related Research
ORIP participates in the NIH-wide Research Supplements to Promote Diversity in Health-Related Research initiative. ORIP/Division of Comparative Medicine Diversity Supplements seek to make significant contributions to the research career development of individuals with disabilities, certain individuals from disadvantaged backgrounds, and individuals from racial and ethnic groups that have been shown by the National Science Foundation to be underrepresented in health-related sciences on a national basis. Since 2021, ORIP has supported four awardees—one baccalaureate degree holder, two doctoral students, and one post-doctoral researcher. These ORIP-supported awardees will contribute to diversifying the biomedical research workforce. - ORIP Individual Career Development and Institutional Training Awards
In 2021–2022, ORIP is supporting 32 Individual Training Grants to veterinary students and veterinarians seeking a Ph.D., as well as to graduate veterinarians, to provide them with research experience to become independent biomedical investigators. Additionally, ORIP is supporting 13 Institutional Research Training Grants to train veterinarians for research careers in comparative medicine and 20 Short-Term Institutional Research Training Grants to provide short-term research training experiences for veterinary students. - ORIP Supports Individual Predoctoral Fellowships to Promote Diversity in Health-Related Research
ORIP accepts applications from highly qualified veterinary students or holders of degrees in veterinary medicine to enhance the diversity of the health-related research workforce (PA-21-052). - ORIP Provides Support for Early-Stage Investigators Using Nonhuman Primate Research Models
This funding opportunity announcement (PAR-20-258) provides early-stage investigators with support and “protected time” (up to 5 years) for intensive, research-focused career development program activities under the guidance of an experienced mentorship team with expertise in both the preclinical application of nonhuman primate models and in translation of the results from such studies to clinical application. Participating NIH Institutes/Centers/Offices (ICOs) include the National Institute of Allergy and Infectious Diseases (lead ICO), National Institute on Aging, National Institute on Alcohol Abuse and Alcoholism, National Center for Complementary and Integrative Health, and ORIP.
ORIP-Supported Research Highlights
- Deletion of Mouse Lysyl Oxidase in Megakaryocytes Affects Bone Properties in a Sex-Dependent Manner
Lysyl oxidase (LOX) is a facilitator of extracellular matrix cross-linking, and the importance of LOX in bone formation has been addressed in both in vitro and in vivo studies. Using newly developed megakaryocyte-specific LOX knockout mice, the researchers show that LOX expressed in these scarce bone marrow cells leads to changes in bone volume and mechanical strength in male mice. No significant changes were observed, however, within the female experimental groups. The authors’ findings suggest that sex hormones could contribute to differences within these dynamics. - Synthetic Protein Circuits for Programmable Control of Mammalian Cell Death
Natural cell death pathways have been shown to eliminate harmful cells and shape immunity. Researchers used synthetic protein-level cell death circuits, collectively termed “synpoptosis” circuits, to proteolytically regulate engineered executioner proteins and mammalian cell death. They show that the circuits direct cell death modes, respond to combinations of protease inputs, and selectively eliminate target cells. This work provides a foundation for programmable control of mammalian cell death. Future studies could focus on programmable control of cell death in various contexts, including cancer, senescence, fibrosis, autoimmunity, and infection. - Controlled Stiffness of Direct-Write, Near-Field Electrospun Gelatin Fibers Generates Differences in Tenocyte Morphology and Gene Expression
Tendinopathy is associated with mobility issues, and the cell–matrix interactions involved in the development of tendinopathy are not fully understood. In this study, researchers fabricated micron-scale fibrous scaffolds that mimic native collagen fiber size and orientation. They reported that matrix metalloproteinase and proteoglycans (possible indicators of tendinopathy) were more upregulated in the presence of high-stiffness fibers than low-stiffness fibers. The authors’ findings suggest that scaffolds can serve as in vitro models for tendinopathy. Additionally, their work suggests that matrix mechanical properties contribute to cell–matrix interactions during tendinopathy formation. - Transcriptome- and Proteome-Wide Effects of a Circular RNA Encompassing Four Early Exons of the Spinal Muscular Atrophy Genes
Spinal muscular atrophy (SMA) is a leading genetic cause of mortality in infants and often results from a deficiency of deletions of or mutations in the SMN1 gene. In this study, researchers report the transcriptome- and proteome-wide effects of overexpression of C2A 2B-3–4, a circular RNA produced by SMN1 and SMN2, in cells. They report that C2A-2B-3–4 is associated with expression of genes associated with chromatin remodeling, transcription, spliceosome function, ribosome biogenesis, lipid metabolism, cytoskeletal formation, cell proliferation, and neuromuscular junction formation. More work is needed to investigate the role of these genes in processes associated with SMA and other pathological conditions, including cancer and male infertility. - Potent HPIV3-Neutralizing IGHV5-51 Antibodies Identified From Multiple Individuals Show L Chain and CDRH3 Promiscuity
Human parainfluenza virus 3 fusion glycoprotein (HPIV3 F), responsible for facilitating viral entry into host cells, is a major target of neutralizing antibodies that inhibit infection. More work is needed to understand these dynamics. Researchers characterized the genetic signatures, epitope specificity, neutralization potential, and publicness of HPIV3-specific antibodies identified across multiple individuals (sex not specified). From this work, they identified 12 potently neutralizing antibodies targeting three nonoverlapping epitopes on HPIV3 F. Six of the antibodies used immunoglobulin heavy variable gene (IGHV5-51). These antibodies used different L chain variable genes (VL) and diverse H chain CDR 3 (CDRH3) sequences. These findings help elucidate the genetic and functional characteristics of HPIV3-neutralizing antibodies and indicate the existence of a reproducible H chain variable–dependent antibody response associated with VL and CDRH3 promiscuity. - Ultrasoft Platelet-Like Particles Stop Bleeding in Rodent and Porcine Models of Trauma
Platelet transfusions are the current standard of care to control bleeding in patients following acute trauma, but their use is limited by short shelf life and limited supply. Immunogenicity and contamination risks also are a concern. Using ultrasoft and highly deformable nanogels coupled to fibrin-specific antibody fragments, researchers developed synthetic platelet-like particles (PLPs) as an alternative for immediate treatment of uncontrolled bleeding. They report that PLPs reduced bleeding and facilitated healing of injured tissue in mice, rat, and swine models (sex not specified) for traumatic injury. These findings can inform further translational studies of synthetic PLPs for the treatment of uncontrolled bleeding in a trauma setting. - Pathogenesis and Virulence of Coronavirus Disease: Comparative Pathology of Animal Models for COVID-19
Researchers have used animal models that can replicate clinical and pathologic features of severe human coronavirus infections to develop novel vaccines and therapeutics in humans. The purpose of this review is to describe important animal models for COVID-19, with an emphasis on comparative pathology. The highlighted species included mice, ferrets, hamsters, and nonhuman primates. Knowledge gained from studying these animal models can help inform appropriate model selection for disease modeling, as well as for vaccine and therapeutic developments. - Epigenetic MLH1 Silencing Concurs With Mismatch Repair Deficiency in Sporadic, Naturally Occurring Colorectal Cancer in Rhesus Macaques
Rhesus macaques serve as a useful model for colorectal cancer (CRC) in humans, but more data are needed to understand the molecular pathogenesis of these cancers. Using male and female rhesus macaques, researchers investigated mismatch repair status, microsatellite instability, genetic mutations, transcriptional differences, and epigenetic alterations associated with CRC. Their data indicate that epigenetic silencing suppresses MLH1 transcription, induces the loss of MLH1 protein, abrogates mismatch repair, and drives genomic instability in naturally occurring CRC in rhesus macaques. This work provides a uniquely informative model for human CRC. - Intestinal Epithelial Adaptations to Vertical Sleeve Gastrectomy Defined at Single-Cell Resolution
Perturbations in the intestinal epithelium have been linked to the pathogenesis of metabolic disease. Bariatric procedures, such as vertical sleeve gastrectomy (VSG), cause gut adaptations that induce robust metabolic improvements. Using a male mouse model, the authors assessed the effects of VSG on different cell lineages of the small intestinal epithelium. They show that Paneth cells display increased expression of the gut peptide Reg3g after VSG. Additionally, VSG restores pathways pertaining to mitochondrial respiration and cellular metabolism, especially within crypt-based cells. Overall, this work demonstrates how adaptations among specific cell types can affect gut epithelial homeostasis; these findings can help researchers develop targeted, less invasive treatment strategies for metabolic disease. - Antibiotic-Induced Gut Dysbiosis Elicits Gut-Brain Axis Relevant Multi-Omic Signatures and Behavioral and Neuroendocrine Changes in a Nonhuman Primate Model
Gut microbiome–mammalian cell interactions influence the development of metabolic, immune-mediated, and neuropsychiatric disorders. Dysbiosis of the gut microbiome has been linked to behavioral characteristics in previous nonhuman primate (NHP) studies, but additional studies using NHPs are necessary to understand microbiota–gut–brain communication. The authors sought to evaluate whether antibiotic-induced gut dysbiosis can elicit changes in gut metabolites and behavior indicative of gut–brain axis disruption in common marmosets of both sexes. For the first time in an NHP model, this study demonstrated that antibiotics induce gut dysbiosis, alter gut metabolites relevant to gut–brain communication, affect neuroendocrine responses in response to stressful stimuli, and change social behavior. - HIV-1 Remission: Accelerating the Path to Permanent HIV-1 Silencing
Current HIV treatment strategies are focused on forced proviral reactivation and elimination of reactivated cells with immunological or toxin-based technologies. Researchers have proposed the use of a novel “block-lock-stop” approach, which entails the long-term durable silencing of viral expression and permanent transcriptional deactivation of the latent provirus. More research is needed to understand the (1) epigenetic architecture of integrated provirus, (2) cell types and epigenetic cell states that favor viral rebound, (3) molecular functions of Tat (a protein that controls transcription of HIV) and host factors that prevent permanent silencing, (4) human endogenous retrovirus silencing in the genome, and (5) approaches to generate defective proviruses. Additionally, community engagement is crucial for this effort. - Baseline Tumor Gene Expression Signatures Correlate With Chemoimmunotherapy Treatment Responsiveness in Canine B Cell Lymphoma
Dogs develop spontaneous diffuse large B cell lymphoma (DLBCL), and veterinary clinical trials have been employed to treat canine DLBCL and to inform clinical trials for humans. Investigators evaluated gene expression in lymph node aspirates from 18 trial dogs and defined good responders as those who relapsed after 90 days, and poor responders as those who relapsed prior to 90 days. They found increased CCND3 correlated with poor prognosis and increased CD36 correlated with good prognosis, as is observed in humans. These findings identify biomarkers that may help guide the choice of chemoimmunotherapy treatment in dogs. - A SACS Deletion Variant in Great Pyrenees Dogs Causes Autosomal Recessive Neuronal Degeneration
ARSACS (autosomal recessive spastic ataxia of Charlevoix-Saguenay) is an early-onset, slowly progressive neurodegenerative disorder. To date, no naturally occurring large animal model has been reported for ARSACS. In this study, the authors describe a novel spontaneous genetic model for SACS-associated neuronal degeneration using Great Pyrenees dogs of both sexes. The canine models described in this study fit closely with the typical early‑onset ARSACS phenotype in humans, and molecular genetic studies demonstrated that these dogs exhibit a deleterious SACS mutation. The clinical and histopathological descriptions of this canine disorder contribute to the description of human ARSACS. - Global Frequency Analyses of Canine Progressive Rod-Cone Degeneration-Progressive Retinal Atrophy and Collie Eye Anomaly Using Commercial Genetic Testing Data
Hundreds of genetic variants associated with canine traits and disorders have been identified; however, the geographic distributions and changes in allele and genotype frequencies over prolonged, continuous periods of time are lacking. This study utilized a large set of genotypes from dogs tested for progressive rod-cone degeneration-progressive retinal atrophy (prcd‑PRA) and collie eye anomaly (CEA). Both diseases exhibited significant differences in genotype frequencies (p=2.7×10-152 for prcd-PRA and 0.023 for CEA) with opposing graphical trends. This study shows that genetic testing informed breeding decisions to produce fewer affected dogs. - Host-Derived Growth Factors Drive ERK Phosphorylation and MCL1 Expression to Promote Osteosarcoma Cell Survival During Metastatic Lung Colonization
Mortality from osteosarcoma is closely linked to lung metastasis, even though the lung appears to be a hostile environment for tumor cells. Using female mice, researchers assessed changes in both host and tumor cells during colonization. Their findings suggest that the mitogen-activated protein kinase (MAPK) pathway is significantly elevated in early and established metastases, which correlates with expression of anti-apoptotic genes (e.g., MCL1). The authors conclude that niche-derived growth factors drive increased MAPK activity and MCL1 expression in osteosarcoma, promoting metastatic colonization. This gene is a promising target for future therapeutic development. - A Gut-Restricted Glutamate Carboxypeptidase II Inhibitor Reduces Monocytic Inflammation and Improves Preclinical Colitis
Many patients with moderate-to-severe inflammatory bowel disease (IBD) do not have adequate disease control. Glutamate carboxypeptidase II (GCPII) offers a promising target for therapeutic development. Researchers generated a class of GCPII inhibitors. They demonstrated that the inhibitor reduced monocytic inflammation in mice and protected against the loss of barrier integrity in primary human colon epithelial air–liquid interface monolayers. Their findings suggest that local inhibition of GCPII could be applied for the development of IBD therapeutics. - Identification of an Anergic BND Cell–Derived Activated B Cell Population (BND2) in Young-Onset Type 1 Diabetes Patients
B cells have been shown to play a role in the pathogenesis of type 1 diabetes (T1D), but the specificity, phenotype, and function of B cells in young-onset T1D is not fully understood. The investigators performed a cross-sectional analysis comparing insulin-reactive to tetanus-reactive B cells in the blood of male and female T1D patients and control participants using mass cytometry. They found elevated insulin-reactive BND2 cells in the blood and pancreatic lymph nodes of young-onset T1D donors. This work suggests that an antigen-specific B cell subset could contribute to the rapid progression of young-onset T1D. - The Eotaxin-1/CCR3 Axis and Matrix Metalloproteinase-9 Are Critical in Anti-NC16A IgE-Induced Bullous Pemphigoid
Bullous pemphigoid is associated with eosinophilic inflammation and circulating and tissue-bound IgG and IgE autoantibodies. Researchers previously established the pathogenicity of anti-NC16A IgE through passive transfer of patient-derived autoantibodies to double-humanized mice. In this study, they characterized the molecular and cellular events that underlie eosinophil recruitment and eosinophil-dependent tissue injury. Their work establishes the eotaxin-1/CCR3 axis and matrix metalloproteinase-9 as key players in the disease and as candidate therapeutic targets for drug development and testing. - Structural Insights into the Broad Protection against H1 Influenza Viruses by a Computationally Optimized Hemagglutinin Vaccine
Influenza is an ongoing public health concern, and computationally optimized broadly reactive antigen (COBRA) hemagglutinin proteins represent a potential strategy for formulating broadly effective influenza vaccines. Researchers determined the crystal structure of COBRA P1, as well as its binding to 1F8, a broadly neutralizing antibody. This work provides valuable insights into the underlying molecular basis for the broad effectiveness of P1, and these insights can be applied to future vaccine designs. - Effect of Viral Strain and Host Age on Clinical Disease and Viral Replication in Immunocompetent Mouse Models of Chikungunya Encephalomyelitis
Chikungunya virus (CHIKV) is associated with neurologic complications, but studies in the central nervous system are challenging to perform in humans. Using a mouse model of both sexes, researchers established the relative severity of neurological disease across multiple stages of neurodevelopment in three strains of CHIKV. The disease was found to be strain dependent, with differences in severity of neurological disease, viral titers in the brain and spinal cord, and proinflammatory gene expression and CD4+ T cell infiltration in the brain. This work provides a mouse model for future studies of CHIKV pathogenesis and the host immune response. - Effects of Pulsatile Intravenous Follicle-Stimulating Hormone Treatment on Ovarian Function in Women with Obesity
By performing intravenous administration of pulsatile recombinant follicle-stimulating hormone, researchers established conditions for effective hypothalamic suppression in women with normal and high body mass index (BMI). In women with obesity, the treatment resulted in E2 and inhibin B levels comparable to those in normal-weight women. This work offers a potential strategy to mitigate some of the adverse effects of high BMI on fertility, assisted reproduction, and pregnancy outcomes. - Canine Models of Charcot-Marie-Tooth: MTMR2, MPZ, and SH3TC2 Variants in Golden Retrievers with Congenital Hypomyelinating Polyneuropathy
Both demyelination and hypomyelination of the nervous system are associated with various clinical diseases. Using whole-genome sequencing, researchers determined the genetic underpinnings of congenital hypomyelinating polyneuropathy in canines of both sexes. These variants genetically describe the first peripheral nervous system–exclusive hypomyelinating polyneuropathies in dogs. By testing for these mutations, breeders can prevent the production of affected offspring. - Phytochemical Compound PB125 Attenuates Skeletal Muscle Mitochondrial Dysfunction and Impaired Proteostasis in a Model of Musculoskeletal Decline
Age-related musculoskeletal decline contributes to disability and cardiometabolic diseases and is linked to impaired mitochondrial function and disrupted proteostasis. Using male and female Hartley guinea pigs, investigators tested a purported phytochemical activator of nuclear factor erythroid 2-related factor 2, which helps promote redox homeostasis, proteome maintenance, and mitochondrial energetics. They reported that the activator, PB125, improved mitochondrial respiration and attenuated disease- and age-related declines in skeletal muscle protein synthesis. This study provides insights for the development of interventions in humans. - Cell-Specific Regulation of Gene Expression Using Splicing-Dependent Frameshifting
Cell type–specific control of gene expression is essential for researchers but is technically challenging. Researchers describe a suite of tools that direct pan-neuronal, excitatory neuron, and photoreceptor-specific gene expression via a splicing-linked expression design (SLED) strategy. SLED can use strong constitutive promoters, without sacrificing precision, by decoupling the tradeoff between promoter strength and selectivity. This approach can be used to selectively deliver fluorescent reporters and calcium indicators to various neuronal subtypes in vivo. SLED can be applied to many different basic and translational research studies. - Progression and Resolution of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) Infection in Golden Syrian Hamsters
Animal models have been essential to studies of COVID-19 pathogenesis, treatment, and prevention of disease. The hamster model offers several key advantages for COVID-19 research; for example, hamsters consistently develop bronchointerstitial pneumonia, which is characterized by acute inflammation, edema, and necrosis and progresses to proliferative interstitial pneumonia. Male and female golden Syrian hamsters were inoculated intranasally with SARS-CoV-2 to track clinical, pathology, virology, and immunology outcomes. The authors report that inoculation reliably induces acute damage to the respiratory tract, followed by a macrophage-dominant pulmonary immune response. These findings can serve as a baseline of SARS-CoV-2 infection outcome measures and could be valuable for determining the efficacy of therapeutic and preventive interventions for COVID-19. - American Alligators Are Capable of West Nile Virus Amplification, Mosquito Infection and Transmission
West Nile virus (WNV) outbreaks have caused large-scale mortalities in farmed American alligators in the southeastern United States, but WNV overwintering is poorly understood. Researchers examined potential ways in which alligators may contribute to the natural ecology of WNV. They reported that alligators are capable of becoming infected by mosquito bite and can serve as a source of infection for mosquitoes. Additionally, they determined that water can serve as a source of infection for alligators but does not easily serve as an intermediate means for transmission between birds and alligators. Taken together, these findings suggest that American alligators are capable of serving as an amplifying host for WNV. Further studies could further examine these species’ contribution to maintenance and overwintering of WNV. - Naturally Occurring Osteochondrosis Latens Lesions Identified by Quantitative and Morphological 10.5 T MRI in Pigs
Juvenile osteochondritis dissecans (JOCD) is a pediatric orthopedic disorder that is associated with pain and gait deficits. JOCD lesions form in the knee, elbow, and ankle joints and can progress to early-onset osteoarthritis. In this study, researchers used a noninvasive magnetic resonance imaging (MRI) method to identify naturally occurring lesions in intact knee and elbow joints of juvenile pigs. This work can be applied to noninvasive identification and monitoring of early JOCD lesions and determination of risk factors that contribute to their progression in children. - Effects of Acute Femoral Head Ischemia on the Growth Plate and Metaphysis in a Piglet Model of Legg-Calvé-Perthes Disease
Legg-Calvé-Perthes disease (LCPD) can lead to permanent deformity of the femoral head and premature osteoarthritis, but the underlying cause remains unknown. More work is needed to determine optimal treatment methods for LCPD. Using a piglet model for LCPD, researchers assessed the effects of acute femoral head ischemia on the proximal femoral growth plate and metaphysis. They reported that alterations to the growth plate zones and metaphysis occurred by 2 days post-ischemia and persisted at 7 days post-ischemia. These findings suggest that growth disruption may occur sooner after the onset of ischemia than researchers had hypothesized previously. - Exosome Cell Origin Affects In Vitro Markers of Tendon Repair in Ovine Macrophages and Tenocytes
The underlying pathogenesis of rotator cuff tendinopathy reflects a combination of intrinsic and extrinsic factors, and recent work suggests that cell-to-cell communication drives the severity of tendon changes. Researchers are interested in the role of extracellular vesicles in tendon mechanical resilience, tissue organization, and anti-inflammatory macrophage phenotype predominance in response to tendon injury. In this study, investigators demonstrated how exosomes differ functionally based on cell source. This work suggests that control of exosome composition could lead to more effective therapies for certain tissues. - A LGR5 Reporter Pig Model Closely Resembles Human Intestine for Improved Study of Stem Cells in Disease
The constant epithelial regeneration in the intestine is the sole responsibility of intestinal epithelial stem cells (ISCs), which reside deep in the intestinal crypt structures. To effectively study ISCs, tools to identify this cell population are necessary. This study validates ISC isolation in a new porcine Leucine-Rich Repeat–Containing G Protein–Coupled Receptor 5 (LGR5) reporter line and demonstrates the use of these pigs as a novel colorectal cancer model. Overall, this novel porcine model provides critical advancement in the field of translational gastrointestinal research. - Phytochemical Compound PB125 Attenuates Skeletal Muscle Mitochondrial Dysfunction and Impaired Proteostasis in a Model of Musculoskeletal Decline
Age-related musculoskeletal decline contributes to disability and cardiometabolic diseases and is linked to impaired mitochondrial function and disrupted proteostasis. Using male and female Hartley guinea pigs, investigators tested a purported phytochemical activator of nuclear factor erythroid 2–related factor 2, which helps promote redox homeostasis, proteome maintenance, and mitochondrial energetics. They reported that the activator, PB125, improved mitochondrial respiration and attenuated disease- and age-related declines in skeletal muscle protein synthesis. This study provides insights for the development of interventions in humans. - Human Hematopoietic Stem Cell Engrafted IL-15 Transgenic NSG Mice Support Robust NK Cell Responses and Sustained HIV-1 Infection
A major obstacle to human natural killer (NK) cell reconstitution is the lack of human interleukin‑15 (IL-15) signaling because murine IL-15 is a poor stimulator of the human IL-15 receptor. Researchers show that immunodeficient NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ (NSG) mice expressing a transgene encoding human IL‑15 (NSG-Tg(IL-15)) have physiological levels of human IL-15 and support long-term engraftment of human NK cells when transplanted with human umbilical cord blood–derived hematopoietic stem cells (HSCs). These mice demonstrate robust and long-term reconstitution with human immune cells but do not develop graft-versus-host disease, allowing long-term studies of human NK cells. The HSC-engrafted mice can sustain HIV-1 infection, resulting in human NK cell responses. This work provides a robust novel model for studying NK cell responses to HIV-1. - Cerebrospinal Fluid Protein Markers Indicate Neuro-Damage in SARS‑CoV‑2-Infected Nonhuman Primates
In this study, researchers examined the proteins expressed in cerebrospinal fluid (CSF) in nonhuman primates (NHPs) to better understand how COVID-19 infection can result in brain pathology, a common outcome. The study found that even in NHPs with minimal or mild COVID-19, CSF proteins were significantly dysregulated compared with uninfected NHPs. Furthermore, the most affected proteins were enriched in the same brain regions that show lesions after COVID‑19 infection, including the cerebral cortex, basal ganglia, and brain stem. Collectively, these regions have wide-ranging control over such crucial functions as cognition, motor control, and breathing, showing how even mild COVID-19 infection can result in significant neurological impairment. - Identification of a Heterogeneous and Dynamic Ciliome During Embryonic Development and Cell Differentiation
Ciliopathies are a class of diseases that arise when the structure or function of the cilium is compromised. To definitively determine the extent of heterogeneity within the ciliome, investigators compared the ciliomes of six distinct embryonic domains. The data comprehensively revealed that about 30% of the ciliome is differentially expressed across analyzed tissues in the developing embryo. Furthermore, upregulation of numerous ciliary genes correlated with osteogenic cell fate decisions, suggesting that changes in the ciliome contribute to distinct functions of cell types in vertebrate species. - In-Depth Virological and Immunological Characterization of HIV-1 Cure After CCR5Δ32/Δ32 Allogeneic Hematopoietic Stem Cell Transplantation
Evidence suggests that CCR5Δ32/Δ32 hematopoietic stem cell transplantation (HSCT) can cure HIV-1, but the immunological and virological correlates are unknown. Investigators performed a longitudinal virological and immunological analysis of the peripheral blood and tissue compartments of a 53-year-old male patient more than 9 years after CCR5Δ32/Δ32 allogeneic HSCT and 48 months after analytical treatment interruption. Sporadic traces of HIV-1 DNA were detected in peripheral T cell subsets and tissue-derived samples, but repeated ex vivo quantitative and in vivo outgrowth assays in humanized mice of both sexes did not reveal replication-competent virus. - Pancreatic Cancer Cells Upregulate LPAR4 in Response to Isolation Stress to Promote an ECM-enriched Niche and Support Tumour Initiation
Understanding drivers of tumor initiation is critical for cancer therapy. Investigators found transient increase of lysophosphatidic acid receptor 4 (LPAR4) in pancreatic cancer cells exposed to environmental stress or chemotherapy. LPAR4 induced tumor initiation, stress tolerance, and drug resistance by downregulating miR-139-5p, a tumor suppressor, and upregulating fibronectin. This study reveals the LPAR4-mediated tumor-initiating niche. - Orthotopic Transplantation of the Full-Length Porcine Intestine After Normothermic Machine Perfusion
Among abdominal solid organ transplants, intestinal transplantation is the least commonly performed and is associated with one of the lowest 5-year patient survival rates. Ischemia-reperfusion injury, which occurs during preservation and transplantation, is associated with postoperative sepsis and allograft rejection. Using a translational porcine model of both sexes, the investigators tested the hypothesis that oxygenated machine perfusion of intestinal grafts would be a feasible approach to improve outcomes after intestine transplantation, as has been demonstrated in other transplanted organs. This study presents the first transplant-validated protocol for successful preservation of intestinal allografts by machine perfusion in a porcine model. - Canine Reference Genome Accuracy Impacts Variant Calling: Lessons Learned from Investigating Embryonic Lethal Variants
With increasingly affordable whole-genome sequencing, hundreds of canine genomes now can be analyzed for embryonic lethal mutations. Investigators examined whole-genome sequence data from 675 dogs of both sexes to investigate for variants with missing homozygosity and high predicted impact. They identified 45 likely embryonic lethal mutations in 32 genes but found that all but one of those were labeled incorrectly and were artifacts associated with a widely utilized canine reference genome. This effect is a major obstacle to studies focusing on loci with high heterozygosity. The investigators propose that by using newer, multiple reference genomes, researchers could reduce artifacts and identify variants more accurately. - Mechanisms and Modeling of Wound Repair in the Intestinal Epithelium
The epithelium’s ability to repair wounded regions is critical to maintaining barrier integrity. In this publication, the authors review in vitro injury models and intestinal cell lines (e.g., IPEC-J2, Caco-2, T-84, HT-29, IEC-6). Controlled artificial wounds allow investigators to explore reparative physiology in cell lines that model diverse aspects of intestinal physiology. More information on these systems (i.e., for creating intestinal injury and understanding the differences in monolayers used for in vitro work) is essential for designing studies that properly capture relevant physiology for the study of intestinal wound repair. - Inhaled Recombinant Human IL-15 in Dogs with Naturally Occurring Pulmonary Metastases from Osteosarcoma or Melanoma: A Phase 1 Study of Clinical Activity and Correlates of Response
Recombinant human interleukin-15 (rhIL-15) has been recognized for its potential as an immunotherapeutic agent for cancer. To date, however, human clinical studies have been limited. Researchers investigated inhaled rhIL-15 in dogs of both sexes with naturally occurring lung metastases from osteosarcoma or melanoma. They observed that both the cytotoxicity of circulating effector cells in the blood while on therapy and the baseline absolute lymphocyte count appear to correlate with clinical benefit. These data support the exploration of combinatorial therapies using inhaled rhIL-15 in both dogs and humans. - A Randomized Controlled Trial of Dietary Rice Bran Intake on Microbiota Diversity, Enteric Dysfunction, and Fecal Secretory IgA in Malian and Nicaraguan Infants
Malnutrition and diarrhea are leading causes of death worldwide in children younger than 5 years of age. Researchers assessed the effects of rice bran supplementation on the intestinal mucosa and local immune system of Malian and Nicaraguan infants at high risk for malnutrition. They reported differences in secretory IgA, markers of environmental enteric dysfunction, and microbiota diversity with supplementation. Their data suggest that rice bran is a promising food supplement that improves mucosal health and microbiota diversity. Long‑term investigations of this supplement in children are needed to understand these effects more fully. - Genetic Characterization of Microsporum canis Clinical Isolates in the United States
Microsporum canis can cause superficial fungal infections in domestic cats and humans. Genotypic characterization of M. canis isolates in the United States had not been conducted previously. In this study, investigators evaluated genetic variants among M. canis isolates from domestic cats in veterinary clinics across the United States. Analysis of microsatellite markers revealed three genetic clusters associated with M. canis. Clinic location and disease severity both were significant predictors of microsatellite variants. Future studies could explore differences in clinical presentation, unknown functions inherent in superficial fungal metabolism, and innovative therapeutic modalities. - Naturally Occurring Osteochondrosis Latens Lesions Identified by Quantitative and Morphological 10.5 T MRI in Pigs
Juvenile osteochondritis dissecans (JOCD) affects the articular–epiphyseal cartilage complex, but its specific pathogenesis is unknown. Noninvasive approaches are needed to characterize the progression of lesions (e.g., osteochondrosis latens [OCL], osteochondrosis manifesta [OCM]) associated with JOCD in vivo. Researchers used magnetic resonance imaging (MRI) to identify naturally occurring OCL lesions at predilection sites in intact joints of juvenile pigs. Histological assessment confirmed the presence of OCL or OCM lesions at each of these sites. Necrotic vascular profiles also were characterized. Future studies in clinical MRI systems are needed to determine the applicability of these methods for early diagnosis in humans. - Characterizing the Metabolic Role of STAT3 in Canine Osteosarcoma
Canine osteosarcoma represents an ongoing therapeutic challenge, and researchers are interested in targeting aberrant signaling pathways associated with the disease. Constitutive activation of STAT3—a transcription factor that mediates numerous biological processes—contributes to survival and proliferation of osteosarcoma cell lines. Investigators characterized metabolic benchmarks associated with STAT3 loss in canine osteosarcoma. Deletion of STAT3 was associated with decreased basal and compensatory glycolysis and diminished invasive capacity. These data support the contribution of STAT3 to aerobic glycolysis and cellular invasive capacity in canine osteosarcoma. Further work is needed to leverage STAT3 as a therapeutic target. - Pharmacogenetic Gene–Drug Associations in Pediatric Burn and Surgery Patients
Simultaneous administration of many medications is common in management of critically ill patients. The researchers investigated drug–drug interactions in these treatments during hospitalization, which might decrease drug efficacy or increase adverse reactions. Genetic and medication data from 30 pediatric burn and surgery patients were analyzed to identify pharmacogene–drug associations. Nineteen patients were identified with predicted altered gene functions. Approximately one-third of the patients tested had functionally impactful genotypes in each of the primary drug metabolizing pathways. This study suggests that the vast variability in drug efficacy is partly due to genetic variants and that pharmacogenetic analysis may help optimize dosing regimens - Structural Basis for Ultrapotent Antibody-Mediated Neutralization of Human Metapneumovirus
No vaccines or therapeutics currently are available for human metapneumovirus (hMPV), a leading cause of morbidity and hospitalization among children worldwide. For vaccine development, the investigators isolated a panel of human monoclonal antibodies (mAbs) against the hMPV fusion (F) protein, the latter of which is the sole target of neutralizing antibodies. They found the majority of the mAbs target diverse epitopes on the hMPV F protein and identified a highly potent mAb, MPV467. They determined the structure of MPV467 in complex with the hMPV F protein, which revealed a complex novel prefusion-specific epitope. These findings revealed the immunodominant antigenic epitopes on the hMPV F protein, identified a mAb therapy for hMPV F disease prevention and treatment, and discovered a prefusion-specific epitope on the hMPV F protein - Losartan Blocks Osteosarcoma-Elicited Monocyte Recruitment and, Combined with the Kinase Inhibitor Toceranib, Exerts Significant Clinical Benefit in Canine Metastatic Osteosarcoma
Osteosarcoma is the most common primary malignant tumor of bone. Overall survival rates for osteosarcoma patients have remained unchanged since improvements associated with the introduction of multidrug chemotherapy protocols in the 1980s. An investigator—under a career development award and training grant, both provided by ORIP—found that a novel therapeutic approach combining losartan and tocerabib was effective in dogs with spontaneous osteosarcoma. This outcome supports further study of this drug combination as a novel therapeutic approach for high-risk patients with metastatic osteosarcoma. - Antibody-Based CCR5 Blockade Protects Macaques from Mucosal SHIV Transmission
Researchers found that competitive inhibition of HIV Env-CCR5 binding via the CCR5-specific antibody Leronlimab protects male and female rhesus macaques against infection following repeated intrarectal challenges with a CCR5-tropic simian-human immunodeficiency virus (SHIV), suggesting that CCR5 blockade with Leronlimab is a promising approach to HIV prophylaxis. - Effects of Persistent Modulation of Intestinal Microbiota on SIV/HIV Vaccination in Rhesus Macaques
Researchers evaluated whether probiotics could improve the immunogenicity and protective efficacy of a simian immunodeficiency virus (SIV)/HIV DNA vaccine plus HIV protein vaccine combination in male rhesus macaques and found that a combination of probiotics and vaccination did not affect rectal SIV/HIV target populations or reduce the rate of heterologous SHIV acquisition during intrarectal challenge. - Psychosocial Stress Alters the Immune Response and Results in Higher Viral Load During Acute SIV Infection in a Pigtailed Macaque Model of HIV
Researchers compared commonly measured parameters of HIV progression between singly and socially housed SIV-infected male pigtailed macaques and found that singly housed animals had a higher viral load in the plasma and cerebrospinal fluid and showed greater CD4+ T cell declines than socially housed macaques. These findings suggest that psychosocial stress could augment the progression of HIV infection.
Progress on Theme 4: Awareness of ORIP Resources and Programs
Programs and Activities Highlights
- ORIP-Supported Research Spotlighted in NIH Research Matters
Research in part supported by the NIH Shared Instrumentation Programs (S10OD025245) was highlighted in NIH Research Matters. A study found that people with two copies of the APOE4 gene variant, called APOE4 homozygotes, almost always develop Alzheimer's disease (AD) pathology by age 55 and experience symptoms about 7 to 10 years earlier than those without the variant. This research suggests that AD in APOE4 homozygotes shares characteristics with other genetically determined forms of AD, emphasizing the need for targeted diagnosis and treatment strategies and more diverse studies across different populations. - American College of Laboratory Animal Medicine Forum 2024
The American College of Laboratory Animal Medicine Forum 2024 was held April 28–May 1, 2024, in Myrtle Beach, South Carolina. The ORIP Director participated in a panel discussion focused on facility and infrastructure improvements to animal programs. As a representative for ORIP, she provided a high-level discussion focused on funding mechanisms, as well as research projects that NIH is interested in funding. - Small Business Fact Sheet
ORIP revised its fact sheet on its Small Business Programs. ORIP reviewed the previous fact sheet for accuracy and updated funding numbers. This is one of several fact sheets that serve as valuable resources for potential investigators to learn about ORIP resources and programs. This fact sheet was published in English and Spanish. - ORIP-Supported Research on NIH Director’s Blog and NIH Research Matters
Research in part supported by the NIH Shared Instrumentation Grant (S10OD028685) was highlighted in both the NIH Director’s Blog and NIH Research Matters. A study found that a specific subtype of the mouth bacterium Fusobacterium nucleatum, called Fusobacterium nucleatum animalis (Fna) C2, may drive colorectal tumor growth. This bacterium's genetic properties allow it to survive in the stomach and infect colorectal tumors, suggesting it could be targeted for colorectal cancer detection and treatment. The findings, reported in Nature, highlight the potential for microbial-based therapies and the need to explore connections between oral health and colorectal cancer. - NIH 2024 INCLUDE Investigators Meeting
The 2024 NIH INCLUDE (INvestigation of Co-occurring conditions across the Lifespan to Understand Down syndromE) Investigators Meeting was held April 15–16, 2024. The event brought together researchers, clinicians, and members of the Down syndrome community to share findings and foster collaboration. Sessions covered a range of topics, including data sharing, clinical trials, and preclinical considerations. ORIP staff served on the meeting planning team, helped review travel award applications, and represented NIH in a breakout session on challenges, gaps, and future directions in the field. - ORIP Grantee Featured in Los Angeles Times
ORIP grantee Dr. Amro Hamdoun’s research (funded by R21OD034075) on stable germline transgenesis in Lytechinus pictus was featured in the Los Angeles Times in an article titled: “Sea urchins made to order: Scripps scientists make transgenic breakthrough.” Hamdoun’s laboratory demonstrated the successful insertion of a piece of foreign DNA for a fluorescent protein that resulted in the first transgenic sea urchin. This work will transform sea urchins into an animal model for understanding neurobiology, developmental biology, and toxicology. - ORIP-Supported Research on NIH Director’s Blog
Research in part supported by an NIH Shared Instrumentation Grant (S10OD023423-01) was highlighted on the NIH Director’s Blog. A new DNA sequencing technique called HiDEF-seq, developed by researchers led by Dr. Gilad Evrony at New York University, can detect early DNA errors before they become permanent mutations, improving our understanding of how mutations arise and potentially helping predict health risks. This method enables highly accurate sequencing of single DNA strands, identifying precursors to mutations and offering new ways to monitor DNA damage from environmental exposures. - BioGRID: Walkthrough
ORIP published a video walkthrough of the Biological General Repository for Interaction Datasets (BioGRID) program in March 2024. BioGRID is an open-access public database that uses structured curation to capture protein, genetic, and chemical interaction data from model organisms and humans. - Aquatic Models Fact Sheet
ORIP updated its Aquatic Models Fact Sheet with up-to-date information in March 2024. ORIP updated previous program numbers and ensured that current notice of funding opportunity due dates are accurate. This fact sheet is one of several fact sheets that are valuable resources for potential investigators to learn about ORIP, ORIP-supported resources, and programs. - ORIP Director’s Blog
In a blog post published in March 2024, the ORIP director describes how ORIP supported critical research infrastructure and technology during the COVID-19 pandemic. ORIP staff evaluated and developed new management plans, preserved and maintained animal resources, continued research center operations to support biomedical communities, and built new resources for COVID-19 research. Overall, the pandemic illustrated the value of critical infrastructure and technology to readily support urgent, emerging, and sustained research needs. - National Science Foundation Site Visit: Gen-4 Engineering Research Centers
An ORIP staff member participated in a site visit in November 2023 to evaluate the annual progress of a National Science Foundation (NSF)–supported Engineering Research Center (ERC). The ERC program supports high-risk, high-payoff research centers focused on advancing engineered systems technology and education through multidisciplinary, cross-sector partnerships. - National Disease Research Interchange Board of Directors Meeting
ORIP presented at the National Disease Research Interchange Board of Directors Meeting, which was held on October 27, 2023. Additionally, ORIP participated in discussions on the Human Tissue and Organ Research Resource and provided insight on ORIP’s strategic directions. Attendance and presentation at the meeting provided a unique opportunity for program staff to promote more awareness about ORIP’s mission, resources, funding opportunities, and strategic goals. - Fall 2023 National Primate Research Center Directors Meeting
The Fall National Primate Research Center (NPRC) Directors meeting, held in October 2023, included program updates from NIH. ORIP also obtained scientific, personnel, and infrastructure briefings regarding each NPRC and participated in a site visit of the California National Primate Research Center. - Research Resource Identifiers Project
ORIP published a promotional video overview of the Research Resource Identifiers (RRID) project in October 2023. The RRID initiative aims to promote research resource identification, discovery, and reuse. It helps solve the challenges of reproducibility, efficiency, and connectivity that result from researchers not being precise when citing biological resources. - Post-Award Procedures and Technical Design Requirements for NIH C06 Projects
ORIP presented financial and program requirements, technical design review process, and other post-award procedures to fiscal year 2023 C06 grant recipients on September 25, 2023. The goal of this presentation was to improve the compliance and management of NIH-funded construction infrastructure programs. - Allocation of Nonhuman Primates (NHPs) for Research
In June 2023, ORIP staff provided an overview to program directors at the National Institute of Allergy and Infectious Diseases (NIAID) regarding the allocation of NHPs from the National Primate Research Centers (NPRCs) and other ORIP-supported colonies. Because the ORIP-supported colonies provide NHPs to a significant number of NIAID-funded studies, there was great interest in better understanding the processes and priorities that are used in allocating animals. This outreach generated several additional rounds of discussion and helped clarify priorities, such as that NIH-supported studies take precedence over industry-funded projects and that investigators’ animal requests must be matched carefully regarding animal characteristics, such as species, age, sex, and immunological status. - Community Assessment Workshop
An ORIP staff member presented on ORIP’s R24 research resources at the Aquatic Germplasm and Genetic Resources Center during the third NIH-HUB Project Meeting on August 5, 2023, which was a part of the Community Assessment Workshop. - Virtual Site Visit to the New York Structural Biology Center
ORIP conducted a virtual site visit to the New York Structural Biology Center on February 1, 2023, to discuss the usage and operations of a facility funded by an ORIP construction award (C06RR015495) before the end of its NIH oversight commitment period. - National Institutional Development Awards (IDeA) Symposium of Biomedical Research Excellence (NISBRE)
ORIP hosted a breakout session featuring the NIH S10 and R24 programs at NISBRE, which was held virtually in December 2022. ORIP also presented on the ORIP/DCI programs and answered participants’ questions. NISBRE is a national scientific meeting that showcases the scientific and training accomplishments of the IDeA program of the National Institute of General Medical Sciences. - America’s Seed Fund Week: One-on-Ones
ORIP program staff reached out to potential small business applicants and invited them to interact with ORIP program officers in July 2022. The potential applicants were afforded the opportunity to ask questions and obtain information about ORIP’s small business programs. Staff from the U.S. Department of Health and Human Services’ Small Business Innovation Research and Small Business Technology Transfer programs also participated. - NIH-Wide Portfolio Analysis of COVID-19 Research Involving Vertebrate Animals in Fiscal Year (FY) 2020 and FY 2021
ORIP completed a portfolio analysis of NIH COVID-19 research involving vertebrate animals conducted during FY 2020 and FY 2021. The effort involved screening, review, and analysis of extramural and intramural research portfolios, excluding contracts. These findings were shared with program staff from other NIH Institutes, Offices, and Centers at the NIH-Wide Workshop on COVID-19 and Animal Model Resources: Lessons Learned and Pandemic Preparedness, held in October 2022. - National Primate Research Centers Interactive Webinars
ORIP staff organized and conducted interactive webinars to train key personnel of the National Primate Research Centers on how to complete and submit their supplementary Research Performance Progress Reports, as well as to introduce a new section for reporting nonhuman primate breeding production based on offspring survival to 30 days. - Animals for Geroscience: Needs for Translational and Preclinical Research
ORIP participated in an interactive discussion panel during a virtual NIH workshop that was held on October 6–7, 2022. The workshop focused the use of animals in basic geroscience-focused research studies across academia, government, and industry settings. Speakers at the workshop examined the challenges and opportunities of studying a variety of animal models for translational geroscience research, as well as the development of interventions that target fundamental aging processes. - America’s SEED Fund: Powered by NIH
The Small Business Administration sponsored the America’s SEED Fund: Powered by NIH meeting in July 2022. ORIP program staff participated in a discussion panel titled “Secrets of a Successful Submission” during the meeting. - America’s Seed Fund Week: One-on-Ones
ORIP program staff reached out to potential small business applicants and invited them to interact with ORIP program officers in July 2022. The potential applicants were afforded the opportunity to ask questions and obtain information about ORIP’s small business programs. Staff from the U.S. Department of Health and Human Services’ Small Business Innovation Research and Small Business Technology Transfer programs also participated. - Overview Video of the Monarch Initiative
An overview video of the Monarch Initiative was posted on ORIP’s website in August 2022. The Monarch Initiative, an ORIP-supported resource, is the biomedical research community’s most comprehensive cross-species phenotype information resource, delivering powerful computational phenotypic tools. - Nonhuman Primate (NHP) Model Systems: State of the Science and Future Needs
ORIP attended the Committee on the State of the Science and Future Needs for NHP Model Systems Public Workshop in August 2022. The workshop was convened by the National Academies of Sciences, Engineering, and Medicine's Committee on the State of the Science and Future Needs for NHP Model Systems. ORIP prepared materials for the Division of Program Coordination, Planning, and Strategic Initiatives Acting Director’s panel presentation highlighting NIH perspectives on this topic. - Sixth HIV Vaccine Trials Network (HVTN) Translational HIV Vaccine Early Stage Investigator (ESI) Conference
ORIP evaluated the success and impact of the sixth HVTN Translational HIV Vaccine ESI Conference, which was held in September 2022. ORIP also contributed to the conference by participating actively in rotating networking and mentoring sessions with ESIs. - TechConnect Small Business Innovation Research (SBIR)/Small Business Technology Transfer (STTR) Spring Innovation Conference
The TechConnect SBIR/STTR Spring Innovation Conference was held at National Harbor in Fort Washington, MD, June 13–15, 2022. Conference attendees met with and listened to SBIR/STTR leaders and experts. Discussions included submission of competitive proposals and leveraging of available resources. Attendees also networked with global industry scouts, investors, and key federal leadership representing the nation’s largest seed fund. ORIP was represented among several NIH program managers who attended the conference. They provided materials and answered questions. Other represented agencies included the Defense Advanced Research Projects Agency, National Aeronautics and Space Administration, National Oceanic and Atmospheric Administration, National Science Foundation, U.S. Air Force, U.S. Navy, U.S. Department of Agriculture, and U.S. Department of Defense - Pilot Centers for Precision Disease Modeling—Comprehensive Functional and Phenotyping Analysis of Disease-Causing Variants in Model Organisms
ORIP participation in the Sequencing Strategies for Population and Cancer Epidemiology Studies (SeqSPACE) webinar series, held May 10, 2022, included presentations on the Pilot U54 Centers of the Precision Disease Models, reports from the U54 principal investigators, and a question-and-answer session. The series was hosted by the National Cancer Institute - Biomedical Research Facilities (PAR-22-088) and Limited Competition: Development and Renovation of Housing, Breeding, and Research Spaces for Existing NIH-supported NHP Colonies (PAR-21-363)
ORIP participated in outreach to fiscal year 2022 C06 applicants (PAR-22-088 and PAR-21-363). They provided an explanation of goals and objectives and facilitated a live question-and-answer session - Quarterly T32 Director’s Consortium Meeting
ORIP offers T32 and T35 Institutional Research Training Grants to encourage veterinarians and veterinary students, respectively, to consider careers in biomedical research. ORIP-supported T32 programs are available at 20 institutions, 17 of which are veterinary schools. Directors of the T32 training grants formed a consortium and meet quarterly. Meeting topics have included reviewing new funding opportunity announcements and updates to NIH policy in relation to Ruth L. Kirschstein National Research Service Award training grants. Training Directors have been discussing the possible formation of a combined T32/T35 Training Director’s Consortium. - U42 Annual Research Performance Progress Report Webinar
ORIP held webinars on February 9 and 17, 2022, to instruct key personnel of ORIP-supported Specific-Pathogen-Free Macaque Breeding Colonies on how to complete their annual Specific-Pathogen-Free Report using a new internet platform. These colonies are maintained free of four or more defined pathogens, and they are well characterized in terms of medical histories and genes that influence infectious disease outcomes. Owing to these attributes, specific-pathogen-free macaques are models for research on HIV vaccines, preventive approaches, and next-generation treatments, for which the specific-pathogen-free breeding colonies are prioritized. ORIP’s new platform facilitates the reporting of colony demographics, animal allocation statistics, and grants supported, thereby enhancing ORIP’s optimal stewardship of this resource for the advancement of AIDS research. - Aquatic Models of Human Diseases Special Webinar: Emerging Aquatic Models and Husbandry
Aquatic animal species, such as zebrafish (Danio rerio), are powerful models for studying human development, behavior, genetics, and disease. Studies indicate that the length, weight, sexual maturation, fecundity, and mortality of zebrafish can vary significantly with different diets, which varies between different facilities and laboratories. The needs and challenges of developing defined diets and optimized feed management strategies for relevant aquatic species used in research were assessed and discussed in the 2018 workshop, “Defined Reference Diets for Zebrafish and Other Aquatic Biomedical Research Models: Needs and Challenges,” convened by ORIP. Because of the workshop, Administrative Supplements for pilot projects of a multiplatform candidate reference diet test for biomedical fish models were awarded to three research resources programs. A summary of the results of those pilots were discussed during the February 2022 webinar, “Aquatic Models of Human Diseases Special Webinar: Emerging Aquatic Models and Husbandry.” - Thirteenth Comparative Medicine Resource Directors Meeting: Innovating, Adapting, and Sustaining Resources for a Dynamic Biomedical Landscape
This virtual meeting held in August 2021 provided a forum for exchange of new information, advances, and ideas and facilitated the development of synergistic working groups, interactions, and collaborations among resources. Session topics included services and research activities, dissemination of specialized knowledge, standard operating procedures, and best practices and support for community outreach. - Pre-Application Webinar for the Research Resource Equipment Program
ORIP held a pre-application webinar for the funding opportunity announcement to support Modern Equipment for Shared-Use Biomedical Research Facilities (PAR-21-326). ORIP staff members assisted potential applicants by explaining the objectives, criteria, and requirements of this funding opportunity announcement. Pre-webinar and live questions from attendees were addressed and answered by program staff. - ORIP Webinar: “What Do Veterinarians (Who Are Also Scientists) Bring to Biomedical Research?”
ORIP's Division of Comparative Medicine supports up to two webinars per year on topics of interest to K01 and F30 awardees. At the fall 2021 meeting, two veterinary scientists discussed their experiences as veterinarians working in biomedical research, as well as the knowledge, skills, and abilities that veterinary scientists bring to the biomedical research enterprise. - 2021 National Small Busines Innovation Research (SBIR) Week—Virtual Event
ORIP participated in this national conference by meeting one-on-one with members of the small business community to provide feedback on project concepts, describe the interest areas of ORIP's Small Business Program, and respond to questions about the SBIR/Small Business Technology Transfer (STTR) application process.
Collaborations with Other NIH Institutes/Centers/Offices
- 2024 Workshop for Early Career Investigators in HIV
The 2024 Workshop for Early Career Investigators in HIV was held virtually on April 24, 2024. Hosted by the Office of AIDS Research (OAR), the workshop provided early-stage investigators, early-career investigators (ECIs), and undergraduate and graduate students with information about OAR and HIV research at NIH, as well as NIH resources that are available to support ECIs. An ORIP staff member served on the organizing committee for this workshop. - Conferences Supported by ORIP and the Office of AIDS Research
The California National Primate Research Center (CNPRC) hosted the 40th Annual Symposium on Nonhuman Primate (NHP) Models for AIDS in October 2023. This symposium offered a key scientific forum for exchanging information and developing collaborations among HIV investigators using NHPs who presented and discussed their work in advancing translation of NHP studies into real-world prophylactic and therapeutic strategies. The HIV Vaccine Trials Network hosted its 7th Annual Translational HIV Vaccine Early-Stage Investigator (ESI) Conference in conjunction with this symposium. This conference provided keynote presentations, learning and skill-building opportunities on priority topics, professional development sessions on grant writing, funding opportunities for and challenges of doing NHP research, advice on working successfully with NIH program officials, information on managing research funding, and opportunities for ESIs to network and interact in translational HIV research. ORIP and the Office of AIDS Research supported these meetings through R13 awards. - Planning Discussion for Human Tissue and Organ Research Resource (HTORR) U42 Notice of Funding Opportunity Reissue in Fall 2024
The HTORR Steering Committee met on August 30, 2023, to address issues planned for the fall meeting. Program staff from ORIP, NEI, NHLBI, NIAID, NIAMS, and NIDDK were represented. Consideration was given to reissue a program announcement and include open competition. Stronger language was recommended related to quality control and innovations. An external advisory committee in the notice of funding opportunity has been incorporated. - Request for Information: Infrastructure for Research in Nonhuman Primates (NOT-OD-23-150)
ORIP and OAR were seeking information on capabilities of commercial organizations, nonprofit organizations, and universities to provide research resources necessary for the conduct of biomedical research in nonhuman primates (NHPs), as well as information on expectations for future needs for NHP resource expectations by the research community between July and September 2023. - Genomics Research to Elucidate the Genetics of Rare diseases (GREGoR) Steering Committee Meeting
An ORIP staff member presented on pilot centers for precision animal models at the GREGoR Steering Committee Meeting, which was held on June 21, 2023. The U54 Pilot Center Program was introduced to the Steering Committee and members of the GREGoR consortium, which is aimed at increasing the proportion of Mendelian disorders with an identified genetic cause through enhanced data sharing; collaboration; and an increased focus on the application of new technologies, sequencing strategies, and analytical approaches. - National Institute on Aging (NIA) Summer Retreat
Two ORIP program staff members presented to NIA leadership and other NIA staff a concept for a new initiative on June 5, 2023. The concept focuses on improving rigor and reproducibility of animal studies by addressing extrinsic factors. - ORIP Small Business Programs
An ORIP staff member presented on ORIP’s Small Business Programs, highlighting the value of supporting technologies to advance research discoveries, at the DPCPSI Deputy Directors Group Meeting on April 12, 2023. - NIH-Wide Workshop on COVID-19 and Animal Model Resources: Lessons Learned and Pandemic Preparedness
In October 2022, ORIP organized an NIH-wide workshop aimed at sharing lessons learned by NIH Institutes, Centers, and Offices (ICOs) about the accessibility of animal models and other research resources needed for the investigation of SARS-CoV-2 and COVID-19. Given the experiences of the ICOs with challenges related to the current global pandemic, the workshop also included discussions of how to best prepare for the next pandemic in terms of facilitating the accessibility of critical animal models and related resources. - The Helping to End Addiction Long-term® Initiative, or NIH HEAL® Initiative, Pain Request for Applications (RFA) Reissue
ORIP’s small business programs signed on to the NIH HEAL Initiative Small Business Innovation Research and Small Business Technology Transfer RFA reissues on pain management and treatment (RFA-NS-23-006 and RFA-NS-23-007) in August 2022. Ten NIH Institutes and Centers had signed on previously to the NIH-wide funding opportunity announcement. - NIH-Supported Resources for Neuroscience Research Using Nonhuman Primates
ORIP participated virtually in a conference on NIH-supported resources for neuroscience research using nonhuman primates, which was held at the University of California, San Diego, in La Jolla, CA. The conference was supported by 1R13NS122451-01A1, which was co‑funded by ORIP - Coordinating Committee on Research on Women's Health (CCRWH) Monthly Meeting: ORIP Updates and Forecasts
An ORIP staff member presented ORIP’s strategic plan progress and program highlights at the Coordinating Committee meeting. - Trans-NIH Nonhuman Primate Resource Planning Working Group Provides Recommendations to Address Domestic Sourcing
The Trans-NIH Nonhuman Primate Resource Planning Working Group, co-chaired by an ORIP member, met in November 2021 to discuss challenges and provide recommendations for the domestic sole sourcing of nonhuman primates necessary for basic and translational research. - Meeting of NIH Staff and the Indian Council of Medical Research
ORIP and other NIH officials met with representatives of the Indian Council of Medical Research in December 2021 to discuss a potential future collaboration that could include a visiting scientist program and the sharing of nonhuman primate models.