Progress on Priority 1: Model Resources to Advance the Study of Human Diseases
Programs and Activities Highlights
- Czech Centre for Phenogenomics Conference 2024
An ORIP staff member represented the International Mouse Phenotyping Consortium–Knockout Mouse Phenotyping Project at an international community phenotyping conference on September 17–18, 2024. He participated in discussions regarding the use of animal models for the development of preclinical drug testing pipelines, as well as genome editing therapeutics. - ORIP Concept Clearance (Reissue): National Primate Research Centers Program
In light of the National Primate Research Centers (NPRCs) Program’s demonstrated success and the critical need to ensure national availability of nonhuman primate (NHP) resources, ORIP requested concept clearance from the Council of Councils on September 12, 2024, to continue its support for the program. The NPRCs Program complements and enables the missions of the other NIH institutes and centers by providing the animals, facilities, expertise, and resources required to enable NHP research in specific disease areas. - Cryopreservation Workshop Session II: Cryopreservation and Development of Sustainable Germplasm Repositories for Aquatic Biomedical Models
The second session of the Cryopreservation and Other Preservation Approaches for Animal Models Workshop was held on September 9–10, 2024. Session II focused on the advancements and future needs of genetic resources of aquatic biomedical models, including zebrafish, Xenopus, Ambystoma, and Xiphophorus. The meeting featured three keynote talks and panel presentations by more than 36 panelists. The workshop participants discussed challenges associated with funding and training, the need for standardized protocols, and the benefits that universal data management systems and training hubs would provide.
ORIP-Supported Research Highlights
- In Vivo Prime Editing Rescues Alternating Hemiplegia of Childhood in Mice
Alternating hemiplegia of childhood (AHC) is a neurodevelopmental disease that can cause involuntary muscle contractions, low muscle tone, paralysis on one side of the body, abnormal eye movements, seizures, and intellectual disability. There currently is no treatment. AHC is caused by a mutation in the gene ATP1A3; three variations of the ATP1A3 gene mutation are responsible for 65% of cases. Researchers used prime editing and base editing tools to correct ATP1A3 gene mutations in cells isolated from AHC patients and two mouse models for AHC (sex not specified). Results showed that physical characteristics of AHC were corrected and that treated mice had an extended lifespan. These findings support the potential use of prime editing and base editing tools to treat this neurological disease. - Determinants of Successful AAV-Vectored Delivery of HIV-1 bNAbs in Early Life
More than 100,000 children are infected with HIV each year through vertical (mother-to-child) transmission. Antiretroviral treatment lapses can occur during postpartum care, which then increases levels of HIV in the mother, resulting in an increased risk of transmission to the infant through breastfeeding. Broadly neutralizing antibodies (bNAbs) defend the host from pathogens and have shown potential as a safe therapy for infants. Gene transfer using adeno-associated virus (AAV) offers an opportunity to provide preventive care for infants at risk of getting HIV. Researchers used an infant rhesus macaque model (sex not specified) for simian immunodeficiency virus (SIV)—equivalent to HIV but in nonhuman primates—to determine whether a single intramuscular injection of AAV-bNAb could protect against SIV vertical transmission. The therapy was more effective in newborn rhesus macaques than in older infants and juveniles, and the newborns also were less likely to develop anti-drug antibodies. Results showed that functional antibodies were present even after 4 years. These findings support the possible use of AAV-bNAb to protect infants from contracting HIV. - Multiplexed Proteomic Biosensor Platform for Label-Free Real-Time Simultaneous Kinetic Screening of Thousands of Protein Interactions
Existing methods for producing functional protein libraries are costly and time-consuming, and they lack real-time kinetic (protein interaction) screening abilities. Researchers developed an automated platform for high-throughput production and screening of a library of proteins on biosensor surfaces. Biosensors are devices that can bind a specific protein in a sample containing many proteins to generate a measurable signal unique to the protein of interest. This allows researchers to complete large-scale kinetic measurements for drug discovery, biomarker identification, and diagnostic development. The platform created by the researchers is known as the Sensor-Integrated Proteome On Chip (SPOC®). SPOC uses nanowells to capture 2,400 proteins at the same time on a single gold biosensor chip. The SPOC biosensor chip can then be analyzed with different techniques to generate kinetic data. The SPOC will allow researchers to understand protein interactions on a large scale for research and clinical applications. - miR-33 Inhibition as a Novel Therapeutic Approach for Treating Muscular Dystrophy
Duchenne muscular dystrophy (DMD) is a devastating disorder caused by changes in the dystrophin gene sequence, which results in the absence of a functional dystrophin protein. Several microRNAs (a type of RNA that can bind to other molecules) can alter DMD by changing gene expression. In this review article, the authors discuss inhibiting microRNAs as a new therapy for DMD. Researchers have shown in a DMD mouse model (sex not specified) that blocking miR-33a/b, a microRNA, can improve muscle regeneration (regrowth of damaged tissue) and reduce DMD symptoms. Anti-microRNA oligonucleotides (AMOs) are short chains of DNA or RNA that block microRNAs. Injection of an AMO that blocks miR-33a/b in the DMD mouse model improved muscle regrowth and increased gene pathways involved in muscle regrowth. These studies highlight the impact of microRNA signaling pathways in DMD and show how they could serve as targets for new therapies to treat the disease. - Distinguishing PEX2 and PEX16 Gene Variant Severity for Mild, Severe, and Atypical Peroxisome Biogenesis Disorders
Peroxisomes are structures in cells that play an important role in metabolism and chemical changes of complex fats. Peroxisomal biogenesis disorders (PBDs) are caused by mutations in peroxin (PEX) genes. PBDs are autosomal recessive diseases—a mutated PEX gene must be passed down from both parents. In patients, symptoms of PBD range from mild to severe multi-organ system defects depending on gene mutations and even different mutations in the same gene. Researchers wanted to understand how different mutations cause the variation in symptoms seen in patients. In fruit flies, the researchers replaced the fly Pex genes with two human PEX genes—PEX2 and PEX16—and different mutant forms of these genes. Researchers found that some mutations caused severe symptoms, such as seizure-like behavior, while others were milder. Introducing a normal functional copy of the human PEX genes into the flies with mutant Pex genes alleviated the symptoms. Further studies with fruit flies will help us understand how different PEX gene mutations affect PBD severity in patients. - Functional Analysis of Pathogenic Variants in LAMB1-Related Leukoencephalopathy Reveals Genotype–Phenotype Correlations and Suggests Its Role in Glial Cells
Cells are surrounded by a matrix, known as the basement membrane, that provides structural support and enhances signaling. Laminin B1 (LAMB1) is a matrix protein in the basement membrane that helps form this supportive structure around cells. Mutations (mistakes in the DNA sequence) in the LAMB1 gene can cause rare neurological disorders. Researchers studied the fruit fly version of the LAMB1 gene, which is LanB1. Using fruit flies, the researchers were able to gain insight into the link between LAMB1 gene mutations and disease symptoms. The LanB1 protein is found in a subset of brain cells, called glia cells, and in the blood–brain barrier. Reducing the amount of LanB1 protein in the blood–brain barrier caused shorter lifespans and movement defects in the fruit flies. Human LAMB1 was not functional in flies, but fly experiments showed that some LanB1 mutations cause severe defects, while others were milder. Tests in human cells suggested some LAMB1 mutations might cause disorders, even in the presence of a normal copy of LAMB1. This study reveals the role of LanB1 in keeping the healthy structure of the fly blood–brain barrier and understanding the consequences of different LAMB1 mutations in humans. - SIV Proviruses Seeded Later in Infection Are Harbored in Short-Lived CD4+ T Cells
HIV can stay dormant for years by mixing its genetic materials into immune cells, making it difficult to remove. It remains unclear which HIV-infected cells survive long term. This study used samples from simian immunodeficiency virus (SIV)–infected macaques (sex not specified) to examine how and when virus-infected cells become part of the long-lived reservoir. Researchers discovered that newer viral sequences were found in short-lived CD4+ T cells, and long-lived cells contained older, more genetically varied viruses. These results suggest that viruses from early infection persist in the long term, whereas newer infections remain in cells that die off quickly, providing insights on future HIV treatment strategies. - Transplantation of Human Kidney Organoids Elicited a Robust Allogeneic Response in a Humanized Mouse Model
Kidney organoids are helping advance studies focused on kidney diseases. However, the use of kidney organoids in studies on tissue transplanted from one organism into another is not well explored. Researchers used a humanized mouse model (sex not specified) that contained a human immune system. Following the transplantation of kidney organoids into the humanized mice, researchers looked at the immune response at days 20 and 30. Results showed that transplantation of kidney organoids caused infiltration of immune cells and an increase in the number of T cells. This study provides a novel platform for bridging the gap between mouse and human studies, which may hasten the development of drugs that reduce tissue rejection in organ transplants. - Caspase-11 Drives Macrophage Hyperinflammation in Models of Polg-Related Mitochondrial Disease
Mitochondria are the energy-producing organelles within cells. Mitochondrial diseases lead to chronic health impairments, which can be worsened by environmental exposures, including bacterial infections. Researchers used a mouse model (sex not specified) to study polymerase gamma (Polg)–related mitochondrial disease. They found that infection with the bacteria Pseudomonas aeruginosa causes macrophages (a type of immune cell) to have an increased response. The response happens through cytokine-mediated increases of caspase-11 and guanylate-binding proteins, which leads to lung inflammation. These findings will help scientists find targets to develop therapies to limit infection- and inflammation-related complications in mitochondrial diseases. - Synaptic Dysregulation in a Mouse Model of GRIN2D Developmental and Epileptic Encephalopathy
Researchers studied a mutation in the GRIN2D gene that is linked to severe developmental delays and epilepsy in children. Using a mouse model (both sexes used), researchers showed that the mutation caused early-onset seizures, abnormal brain activity, and learning impairments. Functional analysis demonstrated increased synaptic activity, leading to heightened hippocampal excitability. These findings highlight how this mutation alters excitatory and inhibitory neuronal signaling in the brain. This work suggests that precision genetic therapy is a promising treatment strategy for patients with mutations in the GRIN2D gene. - Amyotrophic Lateral Sclerosis and Frontotemporal Dementia Mutation Reduces Endothelial TDP-43 and Causes Blood–Brain Barrier Defects
Mutations in the TARDBP gene are linked to neurodegenerative diseases, such as familial frontotemporal dementia (FTD) and amyotrophic lateral sclerosis (ALS). This study showed that the protein TDP-43 is reduced in brain endothelial cells of male and female mice with this mutation—leading to blood–brain barrier (BBB) disruption and causing inflammation, protein buildup, and cognitive issues—mimicking key features of neurodegeneration. These findings suggest that endothelial TDP-43 loss contributes directly to BBB breakdown and disease pathology in ALS-FTD.
Progress on Priority 2: Modern Physical Infrastructure to Accelerate Research Discoveries in Human Health and Diseases
Programs and Activities Highlights
- Site Visit: SUNY Stony Brook
ORIP conducted a virtual site visit to the State University of New York (SUNY) Stony Brook on May 2, 2025. ORIP manages NIH extramural construction programs that fund the construction, renovation, and modernization of research spaces. Grant C06RR029841 provided funding for the construction of the first Animal Biosafety Level 3 Laboratory in the Division of Laboratory Animal Resources at the SUNY Stony Brook Health Sciences Center. Grant C06RR014510 funded the renovation of 25,443 square feet of space on the third floor of the Life Sciences Building at SUNY Stony Brook to create research space for an interdepartmental molecular and cellular biology research group focused on yeast. - Site Visit: Medical University of South Carolina
ORIP conducted a virtual site visit to the Medical University of South Carolina (MUSC) on June 16, 2025. Grant C06RR018823 provided funding to outfit two floors of shell space in the new Darby Children’s Research Institute (CRI) building at MUSC. Renovated research laboratories supported multi-departmental, interdisciplinary research programs in developmental neuroscience and cardiovascular developmental biology and cutting-edge core facilities in lipidomics, cell sorting, tissue engineering, and histology. Grant C06RR015455 funded the upgrade of the sanitization and related support facilities in the existing animal facility on the seventh floor of the Basic Science Building at MUSC and the outfitting of the adjoining shell space on the seventh floor of the CRI into a rodent barrier facility.
ORIP-Supported Research Highlights
- Integrative Multi-Omics Analysis Uncovers Tumor-Immune-Gut Axis Influencing Immunotherapy Outcomes in Ovarian Cancer
Ovarian cancer is the deadliest gynecologic cancer, and effective, long-lasting treatments are needed to maintain a good quality of life for patients with recurrent disease. Researchers conducted a phase 2 clinical trial (NCT02853318) to assess the efficacy of combining pembrolizumab, bevacizumab, and oral cyclophosphamide to treat recurrent ovarian cancer in 40 patients (sex not specified). Their results indicated that the combination regimen extended the patient progression-free survival compared with the single-drug therapies alone. The combination therapy improved quality of life, increased rates of disease control, induced more favorable microbial patterns, and enhanced amino acid and lipid metabolism. These clinical results provide additional treatment options for women with recurrent ovarian cancer. - Lysosomal Dysfunction and Inflammatory Sterol Metabolism in Pulmonary Arterial Hypertension
Dysregulation of lysosomal activity and cholesterol metabolism causes inflammation, but the relevance of these functions to pulmonary arterial hypertension (PAH) is unclear. Researchers examined this topic using both human (both sexes included) and male rodent (rats and mice) endothelial cells (ECs). They found that nuclear receptor coactivator 7 (NCOA7) functions as a homeostatic brake and prevents oxysterol-induced inflammation, EC dysfunction, and PAH. Genetic predisposition to NCOA7 deficiency was driven by single-nucleotide polymorphism, which alters endothelial immunoactivation and correlates with mortality in humans. This study links fundamental lysosomal biology and oxysterol metabolism to EC behavior and may guide potential molecular diagnostics and therapeutics in PAH. - Genetic QT Score as a Predictor of Sudden Cardiac Death in Participants with Sleep-Disordered Breathing in the UK Biobank
Obstructive sleep apnea is characterized by repetitive upper airway collapse. Patients with obstructive sleep apnea have prolonged corrected QT (QTc) intervals during the daytime, which is linked to increased risk for ventricular arrhythmias, sudden cardiac death (SCD), and all-cause mortality. The goal of this study was to evaluate the association between a polygenic risk score for QT prolongation (QTc-PRS), QTc intervals, and mortality in male and female patients enrolled in the UK Biobank. The researchers found that the QTc-PRS was associated with SCD among participants with sleep apnea but not among those without sleep apnea. This work suggests that sleep apnea is a significant modifier of genetic risk. Additionally, Black participants with sleep apnea had a particularly high risk of SCD. - The Saponin Monophosphoryl Lipid A Nanoparticle Adjuvant Induces Dose-Dependent HIV Vaccine Responses in Nonhuman Primates
Researchers tested an HIV vaccine booster using an adjuvant (an ingredient that helps vaccines work better) called saponin monophosphoryl lipid A nanoparticle (SMNP). Using male and female nonhuman primates, researchers found that higher doses of SMNP triggered stronger immune responses, including robust B-cell activation and the production of two neutralizing antibodies (important for long-lasting protection). Only high-dose groups showed significant levels of these antibodies. Findings highlight the importance of dose-dependent potency of vaccines in shaping immune responses. This study suggests SMNP’s potential for use in humans as a next-generation vaccine. - Inferring Drug–Gene Relationships in Cancer Using Literature-Augmented Large Language Models
Scientific literature contains a wealth of information on cancer and cancer drugs. Researchers developed GeneRxGPT, a large language model (LLM)–powered tool that analyzes biomedical literature to uncover drug–gene relationships for cancer treatment. By integrating PubMed data and advanced LLMs, this tool overcomes limitations of static LLMs (e.g., outdated knowledge, misleading results). A case study in liver cancer, supported by an ORIP S10–funded computing cluster, showed a key link between a particular set of mutations and sensitivity to the cancer drug sorafenib, pointing to a new treatment strategy. Designed for accessibility, GeneRxGPT is a promising resource to accelerate cancer drug discovery by helping researchers explore drug–gene interactions. - Engineered Epithelial Curvature Controls Paneth Cell Localization in Intestinal Organoids
Intestinal organoids, laboratory-grown mini-organs that model the intestine, are emerging as a new complementary approach in research. Researchers have developed a new method to design the architecture of intestinal organoids by engineering the curvature of their tissue. Using an ORIP-funded photomanipulation and imaging system combined with a light-sensitive hydrogel, researchers precisely controlled the width and depth of intestinal organoids, mimicking the natural intestinal folds in humans. This structure guided the placement of Paneth cells, which are key to gut health and immunity. By improving the consistency of cell organization, this approach enhances the reproducibility and functionality of organoid models, making them more useful for studying diseases, testing drugs, and advancing restorative medicine. - Giant Polyketide Synthase Enzymes in the Biosynthesis of Giant Marine Polyether Toxins
Researchers identified the “PKZILLAs,” massive polyketide synthase genes in the harmful algae Prymnesium parvum that are responsible for producing prymnesins—large polyether toxins linked to fish kills. PKZILLA-1 and PKZILLA-2 encode enormous proteins, each with more than 90 enzyme domains, producing precursors to A-type and B-type prymnesins. This discovery unveils the long-mysterious biosynthesis of these toxins, providing insights into the genetic and enzymatic mechanisms behind polyether production. It challenges previous size expectations in biological systems, offering new perspectives on polyketide biosynthesis. - Noninvasive Targeted Modulation of Pain Circuits With Focused Ultrasonic Waves
This study explores noninvasive modulation of the anterior cingulate cortex using low-intensity transcranial-focused ultrasound to treat chronic pain. In a randomized crossover trial with 20 male and female patients, 60% experienced significant pain reduction immediately after active stimulation, with sustained effects on days 1 and 7, compared with minimal improvements with sham stimulation. Pain was reduced by 60% immediately post-stimulation and by 43% and 33% on days 1 and 7, respectively. The approach was well tolerated, with only mild, temporary side effects, highlighting its potential as a noninvasive alternative to brain surgery for pain management.
Progress on Priority 3: Innovative Cross-Disciplinary Research Training in Model Systems for Human Health and Diseases
Programs and Activities Highlights
- ORIP and National Eye Institute Training Programs
Training Program Directors from ORIP and the National Eye Institute met on September 16, 2024, to compare their training program outcomes and future strategies. ORIP's Division of Comparative Medicine offers career development support for individuals with D.V.M. or Ph.D. degrees, as well as predoctoral veterinary students.
ORIP-Supported Research Highlights
- Epidemiologically Relevant Phthalate Mixture and Mono(2-Ethyl-5-Hydroxyhexyl) Phthalate Exposure Alter Cell Energy Metabolism in Primary Mouse Granulosa Cells
Many products—including plastic food containers, medical tubing, children’s toys, and personal care products—contain phthalate diesters. Phthalates leach from plastics and negatively affect the environment and female reproductive system. In women, phthalate exposure is linked to endometriosis, polycystic ovarian syndrome, and reduced fertility. The molecular pathways by which phthalate exposure affects ovaries remain understudied. Researchers tested the effects of different phthalate exposures on granulosa cells (cells that support egg cell development). Using female mouse granulosa cells, the researchers found that short-term exposure to phthalates altered the expression of specific genes, Ldha and Glut1, and affected the ability to create energy. These data indicate that phthalate exposure alters metabolism in granulosa cells. - Sphingosine-1-Phosphate Signaling Mediates Shedding of Measles Virus–Infected Respiratory Epithelial Cells
Measles virus (MeV) is an infectious respiratory virus that has a significant global impact and is a major cause of childhood mortality. A single infected person can transmit MeV to nearly 20 other people. Respiratory epithelial cells (cells that line the respiratory system) are the target of MeV infection, and shedding these cells into airborne droplets allows transmission to other people. Researchers used epithelial cells isolated from the tracheas of rhesus macaques (sex not specified) to understand the mechanisms underlying how MeV-infected epithelial cells are shed. Results showed that sphingosine-1-phosphate (S1P) signaling plays a key role in cell shedding. Inhibiting S1P signaling delayed MeV-infected epithelial shedding and increased the amount of virus in the epithelial lining. These findings demonstrate the key role of host cellular responses in MeV infection - A Potential Role for c-MYC in the Regulation of Meibocyte Cell Stress
The integrated stress response (ISR) controls cell survival and promotes apoptosis (a type of cell death) through the protein CHOP during prolonged or severe stress. The ISR has not been evaluated in cancers originating in the glands of the eyelid, such as ocular adnexal sebaceous carcinoma (SebCA). Although SebCA is uncommon, mortality rates of up to 40% have been reported. Researchers studied the role of MYC in regulating the ISR in human meibomian gland epithelial cells (HMGECs) located in the eyelid. Results showed that inhibiting MYC in HMGECs stimulates the ISR, results in a smaller increase in the number of cells, and promotes apoptosis. These data support the role of high MYC as an underlying mechanism for SebCA tumorigenesis. - Activated Polyreactive B Cells Are Clonally Expanded in Autoantibody Positive and Patients with Recent-Onset Type 1 Diabetes
Patients who are prediabetic do not have symptoms but do have autoantibodies (cells that target a normal molecule in the body) present. However, it remains largely unknown how autoreactive B cells affect the development of Type 1 diabetes (T1D). Researchers isolated B cells from the blood of patients with T1D, patients who were prediabetic (AAB), and relatives who were not diabetic and not autoreactive. Results showed that B cells from AAB and T1D patients have altered gene expression in cell signaling and inflammation pathways. These results provide a foundation for future studies focused on identifying biomarkers or creating cell-targeted treatments for T1D. - From In Vitro Development to Accessible Luminal Interface of Neonatal Bovine-Derived Intestinal Organoids
Diarrhea caused by infectious agents in the intestine of newborns remains a major human health concern. Three-dimensional (3D) culture techniques of intestinal epithelial cells have been developed to study host–pathogen interactions as new approach methodologies (NAMs) that complement animal research. With these methods, primary intestinal stem cells from donor intestinal crypts are cultured within an extracellular matrix, which supports the self-organization of the multipotent cells into 3D structures known as intestinal organoids. In this study, the team developed intestinal organoids and organoid-derived single-layer cell cultures to enable research on early-life intestinal function and disease. These organoids captured key aspects of the gastrointestinal lining, how it functions, and the unique roles of different cell types. These models replicate the in vivo intestinal epithelium through their multicellularity, self-replication, and differentiation into mature epithelial cell types and provide a complementary platform for studying human health and disease. - Alterations in Tumor Aggression Following Androgen Receptor Signaling Restoration in Canine Prostate Cancer Cell Lines
Prostate cancer (PCa) ranks second worldwide in cancer-related mortality, but only a few animal models exhibit naturally occurring PCa that recapitulates the symptoms of the disease. Neutered dogs have an increased risk of PCa and often lack androgen receptor (AR) signaling, which is involved in upregulating tumorigenesis but can also suppress aggressive cell growth. In this study, researchers sought to understand more about the role of AR signaling in canine PCa initiation and progression by restoring AR in canine PCa cell lines and treating them with dihydrotestosterone. One cell line exhibited AR-mediated tumor suppression; one cell line showed altered proliferation (but not migration or invasion); and a third cell line exhibited AR-mediated alterations in migration and invasion (but not proliferation). The study highlights the heterogeneous nature of PCa in dogs and humans but suggests that AR signaling might have therapeutic potential under certain conditions.
Progress on Priority 4: Outreach and Awareness of ORIP Resources and Programs
Programs and Activities Highlights
- Oklahoma Medical Research Foundation Construction Supports Inclusive Immunology Research and Proactive Initiatives
ORIP published a research highlight in August 2024 featuring the Oklahoma Medical Research Foundation (OMRF). ORIP’s Extramural Construction Programs supported renovations to facilities for research in molecular immunology and genetics, cardiovascular biology, and autoimmune diseases at OMRF. With this funding, OMRF has been able to expand the breadth of its research, recruit exceptional scientists and trainees, and foster exciting collaborations across Oklahoma and the nation. OMRF has formed partnerships with local Tribes; as a result, OMRF and Tribal clinicians were able to identify better blood markers for the Oklahoma Tribal citizens and change the way that rheumatic diseases are screened for. - S10-Funded Work Featured on Nature Genetics Cover
A recent paper, titled A common flanking variant is associated with enhanced stability of the FGF14-SCA27B repeat locus, was featured on the cover of the July 2024 issue of Nature Genetics. This international study describes a common flanking variant that is associated with enhanced stability of repeat locus. These findings offer insight into the mechanism protecting against tandem repeat expansion, a known cause of more than 40 neurological disorders, including schizophrenia. This work received support through ORIP’s S10 shared instrumentation programs (S10OD026880 and S10OD030463) and was performed using 1,027 samples from the NIH All of Us Research Program.