Grantee Publications

• Neuropeptide S Receptor 1 is a Nonhormonal Treatment Target in Endometriosis
• Previous Exposure to Dengue Virus is Associated with Increased Zika Virus Burden at the Maternal-fetal Interface in Rhesus Macaques
• Effects of Early Daily Alcohol Exposure on Placental Function and Fetal Growth in a Rhesus Macaque Model
• Multiplexed Drug-based Selection and Counterselection Genetic Manipulations in Drosophila
• Neutralizing Antibody Vaccine for Pandemic and Pre-Emergent Coronaviruses
• Blocking α₄β₇ Integrin Delays Viral Rebound in SHIVSF162P3-Infected Macaques Treated with Anti-HIV Broadly Neutralizing Antibodies
• PD-1 Blockade and Vaccination Provide Therapeutic Benefit Against SIV by Inducing Broad and Functional CD8⁺ T Cells in Lymphoid Tissue
• New SHIVs and Improved Design Strategy for Modeling HIV-1 Transmission, Immunopathogenesis, Prevention, and Cure
• IL-21 Enhances Influenza Vaccine Responses in Aged Macaques with Suppressed SIV Infection
• Circulating Integrin α₄β₇+ CD4 T Cells Are Enriched for Proliferative Transcriptional Programs in HIV Infection
• Innate Immunity Stimulation via CpG Oligodeoxynucleotides Ameliorates Alzheimer’s Disease Pathology in Aged Squirrel Monkeys
• Whole-organism 3D Quantitative Characterization of Zebrafish Melanin by Silver Deposition Micro-CT
• MIC-Drop: A Platform for Large-scale in Vivo CRISPR Screens
• SARS-CoV-2 Vaccines Elicit Durable Immune Responses in Infant Rhesus Macaques
• Cryopreservation and Preparation of Thawed Spermatozoa from Rhesus Macaques (Macaca mulatta) for In Vitro Fertilization
• Postpubertal Spermatogonial Stem Cell Transplantation Restores Functional Sperm Production in Rhesus Monkeys Irradiated Before and After Puberty
• Bilateral Visual Projections Exist in Non-teleost Bony Fish and Predate the Emergence of Tetrapods
• Establishing an Immunocompromised Porcine Model of Human Cancer for Novel Therapy Development with Pancreatic Adenocarcinoma and Irreversible Electroporation
• IL-21 and IFNα Therapy Rescues Terminally Differentiated NK Cells and Limits SIV Reservoir in ART-Treated Macaques
• Antibody-Based CCR5 Blockade Protects Macaques from Mucosal SHIV Transmission
• A Yeast Expressed RBD-based SARS-CoV-2 Vaccine Formulated with 3M-052-alum Adjuvant Promotes Protective Efficacy in Non-human Primates
• Western-style Diet Consumption Impairs Maternal Insulin Sensitivity and Glucose Metabolism During Pregnancy in a Japanese Macaque Model
• Early Treatment with a Combination of Two Potent Neutralizing Antibodies Improves Clinical Outcomes and Reduces Virus Replication and Lung Inflammation in SARS CoV-2 Infected Macaques
• Modulation of MHC-E Transport by Viral Decoy Ligands Is Required for RhCMV/SIV Vaccine Efficacy
• In Vitro and in Vivo Functions of SARS-CoV-2 Infection-Enhancing and Neutralizing Antibodies
• A Participant-Derived Xenograft Model of HIV Enables Long-Term Evaluation of Autologous Immunotherapies
• Mitigation of Endemic GI-Tract Pathogen-Mediated Inflammation Through Development of Multimodal Treatment Regimen and Its Impact on SIV Acquisition in Rhesus Macaques
• Protection of Newborn Macaques by Plant-Derived HIV Broadly Neutralizing Antibodies: A Model for Passive Immunotherapy During Breastfeeding
• Recrudescence of Natural Coccidioidomycosis During Combination Antiretroviral Therapy in a Pigtail Macaque Experimentally Infected with Simian Immunodeficiency Virus
• Cell-specific Transcriptional Control of Mitochondrial Metabolism by TIF1γ Drives Erythropoiesis
• Identification of Basp1 as a Novel Angiogenesis-regulating Gene by Multi-model System Studies
• Acoustofluidic Rotational Tweezing Enables High-speed Contactless Morphological Phenotyping of Zebrafish Larvae
• Fluorescence-based Sorting of Caenorhabditis Elegans via Acoustofluidics
• Larval Zebrafish Use Olfactory Detection of Sodium and Chloride to Avoid Salt Water
• Algorithms Underlying Flexible Phototaxis in Larval Zebrafish
• A Platform for Experimental Precision Medicine: The Extended BXD Mouse Family
• A Novel Non-human Primate Model of Pelizaeus-Merzbacher Disease (PMD)
• 'Enhancing' Red Cell Fate through Epigenetic Mechanisms
• Neutralizing Antibody Vaccine for Pandemic and Pre-emergent Coronaviruses
• Loss of Gap Junction Delta-2 (GJD2) Gene Orthologs Leads to Refractive Error in Zebrafish
• Interneuron Origins in the Embryonic Porcine Medial Ganglionic Eminence
• Mineralocorticoid Receptor Blockade Normalizes Coronary Resistance in Obese Swine Independent of Functional Alterations in Kv Channels
• A Symphony of Destruction: Dynamic Differential Fibrinogenolytic Toxicity by Rattlesnake (Crotalus and Sistrurus) Venoms
• Characterization of Axolotl Lampbrush Chromosomes by Fluorescence in Situ Hybridization and Immunostaining
• Advancing Human Disease Research with Fish Evolutionary Mutant Models
• Factor XII Plays a Pathogenic Role in Organ Failure and Death in Baboons Challenged with Staphylococcus aureus
• Systems Vaccinology of the BNT162b2 mRNA Vaccine in Humans
• Modified Adenovirus Prime–Protein Boost Clade C HIV Vaccine Strategy Results in Reduced Viral DNA in Blood and Tissues Following Tier 2 SHIV Challenge
• Interleukin-15 Response Signature Predicts RhCMV/SIV Vaccine Efficacy
• Combining In Vivo Corneal Confocal Microscopy With Deep Learning-Based Analysis Reveals Sensory Nerve Fiber Loss in Acute Simian Immunodeficiency Virus Infection
• Single-cell Protein Activity Analysis Identifies Recurrence-associated Renal Tumor Macrophages
• Thioesterase Superfamily Member 1 Undergoes Stimulus-coupled Conformational Reorganization to Regulate Metabolism in Mice
• Phase Separation Drives Aberrant Chromatin Looping and Cancer Development
• Cryopreservation Method for Drosophila Melanogaster Embryos
• MRI Characteristics of Japanese Macaque Encephalomyelitis (JME): Comparison to Human Diseases
• Evidence in Primates Supporting the Use of Chemogenetics for the Treatment of Human Refractory Neuropsychiatric Disorders
• Rhesus Macaques Build New Social Connections After a Natural Disaster
• Functional Convergence of a Germline-Encoded Neutralizing Antibody Response in Rhesus Macaques Immunized with HCV Envelope Glycoproteins
• Psychosocial Stress Alters the Immune Response and Results in Higher Viral Load During Acute SIV Infection in a Pigtailed Macaque Model of HIV
• Creb5 Establishes the Competence for Prg4 Expression in Articular Cartilage
• Nonhuman Primate Models for SARS-CoV-2 Research: Cryopreservation as a Means to Maintain Critical Models and Enhance the Genetic Diversity of Colonies
• A Chromosome-level Genome of Astyanax Mexicanus Surface Fish for Comparing Population-specific Genetic Differences Contributing to Trait Evolution
• The SARS-CoV-2 Receptor and Other Key Components of the Renin-Angiotensin-Aldosterone System Related to COVID-19 are Expressed in Enterocytes in Larval Zebrafish

• A Novel Tau-Based Rhesus Monkey Model of Alzheimer’s Pathogenesis

• Evaluating a New Class of AKT/mTOR Activators for HIV Latency-Reversing Activity Ex Vivo and In Vivo

• Virus Control in Vaccinated Rhesus Macaques Is Associated with Neutralizing and Capturing Antibodies against the SHIV Challenge Virus but Not with V1V2 Vaccine–Induced Anti-V2 Antibodies Alone

• Immune Variations Throughout the Course of Tuberculosis Treatment and its Relationship with Adrenal Hormone Changes in HIV-1 Patients Co-Infected with Mycobacterium tuberculosis

• Natural Killer Cells Activated through NKG2D Mediate Lung Ischemia-Reperfusion Injury

• Fructose Stimulated De Novo Lipogenesis Is Promoted by Inflammation

• Best Practices for Correctly Identifying Coronavirus by Transmission Electron Microscopy

• A chromosome-level genome of Astyanax mexicanus surface fish for comparing population-specific genetic differences contributing to trait evolution

• A Novel Tau-Based Rhesus Monkey Model of Alzheimer’s Pathogenesis

• A pulsatile release platform based on photo-induced imine-crosslinking hydrogel promotes scarless wound healing

• Resident Memory T Cells Form during Persistent Antigen Exposure Leading to Allograft Rejection

• Metabolomics analysis of follicular fluid coupled with oocyte aspiration reveals importance of glucocorticoids in primate periovulatory follicle competency

• The giant axolotl genome uncovers the evolution, scaling, and transcriptional control of complex gene loci

• Cytomegaloviral Determinants of CD8⁺ T Cell Programming and RhCMV/SIV Vaccine Efficacy

• Sensitive Tracking of Circulating Viral RNA Through All Stages of SARS-CoV-2 Infection

• Polyfunctional Tier 2–Neutralizing Antibodies Cloned Following HIV-1 Env Macaque Immunization Mirror Native Antibodies in a Human Donor
• Autologous Transplant Therapy Alleviates Motor and Depressive Behaviors in Parkinsonian Monkeys
• Trim-Away Mediated Knock Down Uncovers a New Function for Lbh During Gastrulation of Xenopus laevis
• Immunogenic BNT162b Vaccines Protect Rhesus Macaques from SARS-CoV-2
• Modified Vaccinia Ankara Vector-Based Vaccine Protects Macaques from SARS-CoV-2 Infection, Immune Pathology and Dysfunction in the Lung
• Lung Expression of Human Angiotensin-Converting Enzyme 2 Sensitizes the Mouse to SARS-CoV-2 Infection
• SARS-CoV-2 Induces Robust Germinal Center CD4 T Follicular Helper Cell Responses in Rhesus Macaques
• Myelin‐specific T Cells in Animals with Japanese Macaque Encephalomyelitis
• Antibody-Mediated Depletion of Viral Reservoirs is Limited in SIV-Infected Macaques Treated Early With Antiretroviral Therapy
• A Modular Master Regulator Landscape Controls Cancer Transcriptional Identity
• Increased Proviral DNA in Circulating Cells Correlates with Plasma Viral Rebound in SIV-Infected Rhesus Macaques after Antiretroviral Therapy Interruption
• Thresholds for Post-Rebound SHIV Control after CCR5 Gene-Edited Autologous Hematopoietic Cell Transplantation
• Severely Ill COVID-19 Patients Display Impaired Exhaustion Features in SARS-CoV-2-Reactive CD8+ T Cells
• Endogenous Zebrafish Neural Cre Drivers Generated by CRISPR/Cas9 Short Homology Directed Targeted Integration
• Deploying MMEJ using MENdel in Precision Gene Editing Applications for Gene Therapy and Functional Genomics

 

 

Neuropeptide S Receptor 1 is a Nonhormonal Treatment Target in Endometriosis

Tapmeier, Thomas T. et al., Science Translational Medicine, 25 August 2021; Volume 13, Issue 608

https://www.science.org/doi/10.1126/scitranslmed.abd6469?_ga=2.257127624.2111692662.1631560700-993035711.1631035339

Investigators analyzed genetic sequences of humans (n=32 families) and pedigree rhesus macaques (n=849) with spontaneous endometriosis to uncover potential targets for treatment. Target associations indicated a common insertion/deletion variant in NPSR1, the gene encoding neuropeptide S receptor 1. Immunocytochemistry, RT-PCR, and flow cytometry experiments indicated NPSR1 was expressed in the glandular epithelium of eutopic and ectopic endometrium. In a mouse model of endometriosis, an inhibitor if NPSR1-mediated signaling blocked proinflammatory TNFα release, monocyte chemotaxis, and inflammatory cell infiltrate. Further studies in nonhuman primates are needed; however, these results provide support for a nonhormonal treatment of endometriosis.
Supported by ORIP (R24OD011173 and P51OD011106).

 

Previous Exposure to Dengue Virus is Associated with Increased Zika Virus Burden at the Maternal-fetal Interface in Rhesus Macaques

Crooks, Chelsea M. et al., PLoS (Public Library of Science) Pathogens Neglected Tropical Diseases, 30 July 2021

https://journals.plos.org/plosntds/article?id=10.1371/journal.pntd.0009641

Pre-existing immunity to dengue virus (DENV) results in antibody-dependent enhancement (ADE) among DENV serotypes; Zika virus (ZIKV) has homology with DENV suggesting pre-existing DENV immunity may have an impact on ZIKV pathogenesis during pregnancy. In a rhesus macaque model, prior DENV-2 exposure resulted in a higher burden of ZIKV viral RNA in maternal-fetal interface tissues as compared to DENV-naive macaques. However, pre-existing DENV immunity had no detectable impact on ZIKV replication kinetics in maternal plasma; all pregnancies progressed to term without adverse outcomes at delivery. Investigating potential ADE in pregnant women is important as vaccines against DENV and ZIKV are developed.
Supported by ORIP (P51OD011106) and NIAID.

 

Effects of Early Daily Alcohol Exposure on Placental Function and Fetal Growth in a Rhesus Macaque Model

Lo, Jamie O. et al., American Journal of Obstetrics and Gynecology, 5 August 2021

https://www.sciencedirect.com/science/article/pii/S0002937821008309?via%3Dihub

In a rhesus macaque model of chronic prenatal alcohol exposure, daily consumption during early pregnancy significantly diminished placental perfusion at mid to late gestation and significantly decreased the oxygen supply to the fetal vasculature throughout pregnancy. These findings were associated with the presence of microscopic placental infarctions. Although placental adaptations may compensate for early environmental perturbations to fetal growth, placental blood flow and oxygenation were reduced, consistent with the evidence of placental ischemic injury that persisted throughout pregnancy.
Supported by ORIP (P51OD011092), NICHD, and NIAAA.

 

Multiplexed Drug-based Selection and Counterselection Genetic Manipulations in Drosophila

Matinyan, Nick, et al., Cell Reports, 36, 109700, 14 September 2021

https://www.cell.com/cell-reports/pdf/S2211-1247(21)01147-5.pdf

Many highly efficient methods exist which enable transgenic flies to contribute to diagnostics and therapeutics for human diseases. A recent publication, “Multiplexed drug-based selection and counterselection genetic manipulations in Drosophila”, brings the transgenic efficiency up to a new level. Researchers described a drug-based genetic platform with four selection and two counterselection markers increasing transgenic efficiency by > 10-fold compared to established methods in flies. Researchers also developed a plasmid library to adapt this technology to other model organisms. This highly efficient transgenic approach significantly increases the power of not only D. melanogaster but many other model organisms for biomedical research.

 

Neutralizing Antibody Vaccine for Pandemic and Pre-Emergent Coronaviruses

Saunders, Kevin O. et al., Nature, 2021 June;594(7864):553-559

https://doi.org/10.1038/s41586-021-03594-0

SARS-CoV-2 is a new member of the betacoronavirus (beta-CoV) genus, which also includes two common mild beta-CoVs and the life-threatening SARS-CoV-1 and MERS-CoV. Vaccines that elicit protective immunity against SARS-CoV-2 and beta-CoVs that circulate in animals could prevent future pandemics. Researchers designed a novel 24-mer SARS-CoV-2 receptor binding domain-sortase A conjugated nanoparticle vaccine (RBD-scNP). Investigators demonstrated that the immunization of macaques with RBD-scNP, and adjuvanted with 3M-052 and alum, elicits cross-neutralizing antibody responses against bat coronaviruses, SARS-CoV, and multiple SARS-CoV-2 variants of concern. This pioneering approach serves as a multimeric protein platform for the further development of generalized anti-beta-CoV vaccines.
Supported by ORIP (U42OD021458), NIAID, and NCI.

 

Blocking α₄β₇ Integrin Delays Viral Rebound in SHIV_SF162P3-Infected Macaques Treated with Anti-HIV Broadly Neutralizing Antibodies

Frank, Ines et al., Science Translational Medicine, 18 August 2021, Volume 13, Issue 607:eabf7201

https://doi.org/10.1126/scitranslmed.abf7201

To explore therapeutic potentials of combining anti-HIV broadly neutralizing antibodies (bNAbs) with α₄β₇ integrin blockade using the monoclonal antibody Rh-α4β7, investigators treated SHIV_SF162P3-infected, viremic macaques with bNAbs only or bNAbs and Rh-α₄β₇. Treatment with bNAbs alone decreased viremia below 200 copies/ml in eight out of eight macaques, but seven of the monkeys rebounded within 3 weeks. In contrast, three of six macaques treated with both Rh-α4β7 and bNAbs maintained viremia below 200 copies/ml for 21 weeks, whereas three of those monkeys rebounded after six weeks. These findings suggest that α₄β₇ integrin blockade may prolong virologic control by bNAbs in SHIV_SF162P3-infected macaques.
Supported by ORIP (P51OD011104, U42OD010568, U42OD024282, P40OD028116), NIAID, and NCI.

 

PD-1 Blockade and Vaccination Provide Therapeutic Benefit Against SIV by Inducing Broad and Functional CD8⁺ T Cells in Lymphoid Tissue

Rahman, Sheikh Abdul et al., Science Immunology, 3 September 2021, Volume 6, Issue 63:eabh3034.

https://doi.org/10.1126/sciimmunol.abh3034

Effective HIV therapies must induce functional CD8⁺ T cells and clear latent viral reservoirs during antiretroviral therapy (ART). Using a rhesus macaque model, researchers showed that therapeutic vaccination under ART using a CD40L plus TLR7 agonist-adjuvanted DNA/modified vaccinia Ankara vaccine regimen induced robust SIV-specific CD4⁺ and CD8⁺ T cell responses. Addition of an anti-PD-1 antibody to the SIV vaccine increased cytotoxic CD8⁺ T cells in lymph nodes after ART interruption, correlating to the control of virus and prolonged survival compared with the vaccine alone. Thus, combining immune checkpoint blockade with vaccination may be a promising avenue toward an HIV cure.
Supported by ORIP (P51OD011132), NIAID, and Emory University Integrated Cellular Imaging Microscopy Core.

 

New SHIVs and Improved Design Strategy for Modeling HIV-1 Transmission, Immunopathogenesis, Prevention, and Cure

Li, Hui et al., Journal of Virology, 3 March 2021;95(11):e00071-21

https://doi.org/10.1128/JVI.00071-21

Researchers knew that substitution of HIV-1 Env residue 375-serine by aromatic residues enhances binding to rhesus CD4 enabling primary HIV-1 Envs to support replication as simian-human immunodeficiency virus (SHIV) chimeras in rhesus monkeys. The investigators constructed SHIVs containing 10 primary Envs corresponding to HIV-1 subtypes A, B, C, AE, and AG. Only one with histidine at Env375 replicated efficiently in rhesus cells. Replacement of wildtype Env375 residues by tryptophan, tyrosine, phenylalanine, or histidine in the other 9 SHIVs led to efficient replication. These new SHIVs transmit via mucosal routes like HIV-1 and have use for vaccine testing in nonhuman primates.
Supported by ORIP (U42OD021458, P40OD012217), NIAID, NCI, and Bill & Melinda Gates Foundation.

 

IL-21 Enhances Influenza Vaccine Responses in Aged Macaques with Suppressed SIV Infection

Kvistad, Daniel et al., JCI Insight, 7 September 2021, online ahead of print.

https://doi.org/10.1172/jci.insight.150888

Aging with HIV is associated with low-grade systemic inflammation, immune senescence, and impaired antibody (Ab) responses to vaccines such as influenza (flu). Researchers investigated the role of Interleukin (IL)-21, a CD4 T follicular helper cell regulator, on flu vaccine Ab response in rhesus macaques in the context of age and controlled simian immunodeficiency virus (SIV) mac239 infection. They found that IL-21 enhanced flu vaccine-induced Ab responses in SIV+ (anti-retroviral therapy-suppressed) aged rhesus macaques, adjuvanting the flu vaccine by modulating lymph node germinal center activity. Thus, strategies to supplement IL-21 in aging might improve vaccine responses in people aging with HIV.
Supported by ORIP (R24OD010947) and NIAID.

 

Circulating Integrin α₄β₇⁺ CD4 T Cells Are Enriched for Proliferative Transcriptional Programs in HIV Infection

Lakshmanappa, Yashavanth S. et al., FEBS (Federation of European Biochemical Societies) Letters, 5 September 2021;595(17):2257-2270

https://doi.org/10.1002/1873-3468.14163

HIV preferentially infects α₄β₇⁺ CD4 T cells, forming latent reservoirs that contribute to HIV persistence, yet the properties of α₄β₇⁺ CD4 T cells are poorly understood. Investigating HIV-infected humans and SHIV-infected rhesus macaques, investigators demonstrated that α₄β₇⁺ CD4 T cells in blood are enriched for genes regulating cell cycle progression and cellular metabolism. In contrast, rectal α₄β₇⁺ CD4 T cells exhibited a core tissue-residency gene expression program. These features were conserved across primate species, suggesting that the tissue environment influences memory T-cell transcriptional networks. These findings provide an important foundation for understanding the role of α₄β₇ in HIV infection.
Supported by ORIP (K01OD023034, R24OD010976) and NIAID.

 

Innate Immunity Stimulation via CpG Oligodeoxynucleotides Ameliorates Alzheimer’s Disease Pathology in Aged Squirrel Monkeys

Patel, Akash G., et al., Brain, A Journal of Neurology, July 2021, Volume 144, Issue 7

https://academic.oup.com/brain/article/144/7/2146/6297373?login=true

Alzheimer's disease is the only illness among the top 10 causes of death for which there is no disease-modifying therapy. The authors have shown in transgenic Alzheimer's disease mouse models that harnessing innate immunity via TLR9 agonist CpG oligodeoxynucleotides (ODNs) modulates age-related defects associated with immune cells and safely reduces amyloid plaques, oligomeric amyloid-β, tau pathology, and cerebral amyloid angiopathy (CAA). They used a non-human primate model of sporadic Alzheimer's disease pathology that develops extensive CAA-elderly squirrel monkeys. They demonstrate that long-term use of Class B CpG ODN 2006 induces a favorable degree of innate immunity stimulation.  CpG ODN 2006 has been well established in numerous human trials for a variety of diseases. This evidence together with their earlier research validates the beneficial therapeutic outcomes and safety of this innovative immunomodulatory approach.
Supported by ORIP (P40 OD010938), NINDS, NIA, NCI

 

Whole-organism 3D Quantitative Characterization of Zebrafish Melanin by Silver Deposition Micro-CT

Katz, Spencer R., et al, Elife, 16 September 2021;10:e68920. PMC8445617

https://www.biorxiv.org/content/10.1101/2021.03.11.434673v1

This research team combined micro-CT with a novel application of ionic silver staining to characterize melanin distribution in whole zebrafish larvae. The resulting images enabled whole-body, computational analyses of regional melanin content and morphology. Normalized micro-CT reconstructions of silver-stained fish consistently reproduced pigment patterns seen by light microscopy and allowed direct quantitative comparisons of melanin content. Silver staining of melanin for micro-CT provides proof-of-principle for whole-body, 3D computational phenomic analysis of a specific cell type at cellular resolution. Advances such as this in whole-organism, high-resolution phenotyping provide superior context for studying the phenotypic effects of genetic, disease, and environmental variables.
Supported by ORIP (R24 OD018559)

 

MIC-Drop: A Platform for Large-scale in Vivo CRISPR Screens

Parvez, Saba et al., Science, 3 September 2021;373(6559):1146-1151

https://pubmed.ncbi.nlm.nih.gov/34413171/

CRISPR screens in animals are challenging because generating, validating, and keeping track of large numbers of mutant animals is prohibitive. These authors introduce Multiplexed Intermixed CRISPR Droplets (MIC-Drop), a platform combining droplet microfluidics, single-needle en masse CRISPR ribonucleoprotein injections, and DNA barcoding to enable large-scale functional genetic screens in zebrafish. In one application, they showed that MIC-Drop could identify small-molecule targets. Furthermore, in a MIC-Drop screen of 188 poorly characterized genes, they discovered several genes important for cardiac development and function. With the potential to scale to thousands of genes, MIC-Drop enables genome-scale reverse genetic screens in model organisms.
Supported by ORIP (R24 OD017870), NIGMS, NHLBI

 

SARS-CoV-2 Vaccines Elicit Durable Immune Responses in Infant Rhesus Macaques

Garrido, Carolina et al., Science Immunology, 15 June 2021; Vol. 6, Issue 60.

https://immunology.sciencemag.org/content/6/60/eabj3684

The immunogenicity of two SARS-CoV-2 vaccines was evaluated in both sexes of infant rhesus macaques (n=8/group). Neither vaccine, stabilized prefusion SARS-CoV-2 S-2P spike (S) protein encoded by mRNA encapsulated in lipid nanoparticles or the purified S protein mixed with 3M-052, a synthetic TLR7/8 agonist in a squalene emulsion, induced adverse effects. Both elicited high magnitude neutralizing antibody titers peaking at week 6. S-specific T cell responses were dominated by IL-17, IFN-γ, or TNF-α. Antibody and cellular responses were stable through week 22. These data provide proof-of concept for a pediatric SARS-CoV-2 vaccine with the potential for durable immunity to decrease transmission of COVID-19.
Supported by ORIP (P51OD011107), NIAID, and NCI.

 

Cryopreservation and Preparation of Thawed Spermatozoa from Rhesus Macaques (Macaca mulatta) for In Vitro Fertilization

De Carvalho, Fernanda M. et al., 23 May 2021; Journal of the American Association for Laboratory Animal Science.

https://www.ingentaconnect.com/content/aalas/jaalas/pre-prints/content-jaalas-20-000028

Optimizing procedures for cryopreservation and subsequent thawing for rhesus macaques is required to prevent cryodamage that negatively impacts artificial insemination and in vitro fertilization rates. Investigators systematically assessed two cryopreservation methods and four recovery methods in three interdependent experiments. Results demonstrated that slow-freezing, coupled with density gradient centrifugation provided the highest efficacy in functional sperm for in vitro use. Additional studies are required to further optimize sperm cryopreservation in rhesus macaques.
Supported by ORIP (P51OD011092).

 

Postpubertal Spermatogonial Stem Cell Transplantation Restores Functional Sperm Production in Rhesus Monkeys Irradiated Before and After Puberty

Shetty, Gunapala et al., 06 May 2021; Andrology.

https://onlinelibrary.wiley.com/doi/10.1111/andr.13033

Cancer treatment of prepubertal patients impacts future fertility due to the abolition of spermatogonial stem cells (SSCs). Prepubertal rhesus monkeys (n=6) were unilaterally castrated, and the remaining testes irradiated twice to insure loss of SSCs; the animals were treated with a vehicle or GnRH antagonist for 8 weeks (n=3/treatment). The cryopreserved prepubertal testicular tissue was allergenically transplanted into the intact testes of the monkeys after puberty. Recovery of viable donor epididymal sperm was observed in half the monkeys. These results illustrate that sperm production can be restored in primates by transplantation of testicular cells from cryopreserved untreated prepubertal testes into seminiferous tubules of the remaining testes.
Supported by ORIP (P51OD011092), NICHD, and NCI.

 

Bilateral Visual Projections Exist in Non-teleost Bony Fish and Predate the Emergence of Tetrapods

Vigouroux, Robin J., et al. Science. 09 April 2021; 372(6538):150-156.

https://science.sciencemag.org/content/372/6538/150

In most vertebrates, camera-style eyes contain retinal ganglion cell neurons that project to visual centers on both sides of the brain. However, in fish, ganglion cells were thought to innervate only the contralateral side, suggesting that bilateral visual projections appeared in tetrapods. Here, Vigouroux et al., showed that bilateral visual projections exist in non-teleost fishes and that the appearance of ipsilateral projections does not correlate with terrestrial transition or predatory behavior. However, overexpression of human ZIC2 induces ipsilateral visual projections in zebrafish. Therefore, the existence of bilateral visual projections likely preceded the emergence of binocular vision in tetrapods.
Supported by ORIP (R01 OD011116).

 

Establishing an Immunocompromised Porcine Model of Human Cancer for Novel Therapy Development with Pancreatic Adenocarcinoma and Irreversible Electroporation

Hendricks-Wenger, Alissa et al., Scientific Reports. 07 April 2021; 11(1):7584 (PMC8027815).

https://www.nature.com/articles/s41598-021-87228-5

Efficacious interventions to treat pancreatic cancer lack a preclinical model to recapitulate patients' anatomy and physiology. The authors developed RAG2/IL2RG deficient pigs using CRISPR/Cas9 with the novel application of cancer xenograft studies of human pancreatic adenocarcinoma. These pigs were successfully generated using on-demand genetic modifications in embryos. Human Panc01 cells injected into the ears of RAG2/IL2RG deficient pigs demonstrated 100% engraftment. The electrical properties and response to irreversible electroporation of the tumor tissue were found to be similar to excised human pancreatic cancer tumors. This model will be useful to bridge the gap of translating therapies from the bench to clinical application.
Supported by ORIP (R21OD027062), NIBIB, NCI

 

IL-21 and IFNα Therapy Rescues Terminally Differentiated NK Cells and Limits SIV Reservoir in ART-Treated Macaques

Harper, Justin et al., Nature Communications. 2021 May 17; 12:2866.

https://doi.org/10.1038/s41467-021-23189-7

Nonpathogenic simian immunodeficiency virus (SIV) infections in natural hosts, such as vervet monkeys, are characterized by a lack of gut microbial translocation, robust secondary lymphoid natural killer cell responses, and limited SIV dissemination in lymph node B-cell follicles. Using antiretroviral therapy-treated, SIV-infected rhesus monkeys—a pathogenic model—researchers showed that interleukin-21 and interferon alpha therapy generate terminally differentiated blood natural killer cells with potent human leukocyte antigen-E-restricted activity in response to SIV envelope peptides. The correlated reduction of replication-competent SIV in lymph node demonstrates that vervet-like natural killer cell differentiation can be rescued in rhesus monkeys to promote viral clearance.
Supported by ORIP (P51OD011132, R24OD010947), NIAID, NCI, French National Agency for Research on AIDS and Viral Hepatitis, and Fondation J. Beytout.

 

Antibody-Based CCR5 Blockade Protects Macaques from Mucosal SHIV Transmission

Chang, Xiao L. et al., Nature Communications. 2021 Jun 7; 12:3343.

https://doi.org/10.1038/s41467-021-23697-6

The efficacy of antiretroviral therapy (ART) as pre-exposure prophylaxis against HIV is hindered by incomplete patient adherence and ART-resistant variants. Researchers found that competitive inhibition of HIV Env-CCR5 binding via the CCR5-specific antibody Leronlimab protects rhesus macaques against infection following repeated intrarectal challenges with a CCR5-tropic simian-human immunodeficiency virus (SHIV_SF162P3). Biweekly injection of Leronlimab at 50 mg/kg provided complete protection from SHIV infection. Tissue biopsies from protected macaques post-challenge revealed complete CCR5 receptor occupancy and an absence of viral DNA. After Leronlimab washout, transfer of hematologic cells into naïve monkeys did not transmit infection, supporting the initiation of clinical trials.
Supported by ORIP (P51OD011092, K01OD026561, P40OD028116) and NIAID.

 

A Yeast Expressed RBD-based SARS-CoV-2 Vaccine Formulated with 3M-052-alum Adjuvant Promotes Protective Efficacy in Non-human Primates

Pino, Maria et al., Science Immunology, 15 July 2021; Volume 6, Issue 61

https://immunology.sciencemag.org/content/6/61/eabh3634

Using a rhesus macaque model (n=5 males per group), investigators tested a receptor binding domain (RBD) recombinant protein formulation COVID vaccine candidate combined with an aluminum-based formulation of 3M’s Toll-like receptor 7 and 8 agonist 3M-052 (3M-052/Alum) and found the RBD+3M-052/Alum formulation produced a superior overall immune response than RBD+alum alone as demonstrated by higher SARS-CoV-2 neutralizing antibodies, improved Th1 biased CD4+T cell reactions, and increased CD8+ T cell responses. Collectively, these data suggest that the RBD+3M-052-alum formulation provides robust immune responses against SARS-CoV-2 and supports the development of this potential effective and easy to scale COVID vaccine candidate.
Supported by ORIP (P51OD011132) and NIAID.

 

Western-style Diet Consumption Impairs Maternal Insulin Sensitivity and Glucose Metabolism During Pregnancy in a Japanese Macaque Model

Elsakr, Joseph M. et al., Scientific Reports, 21 June 2021; Volume 11, Issue 12977

https://www.nature.com/articles/s41598-021-92464-w

Using a Japanese macaque model, investigators assessed the metabolic effects of obesity and a calorically dense, Western-style diet (WSD; 36.3% fat), either alone or together, on maternal glucose tolerance and insulin levels in dams during pregnancy (n = 95 females followed over multiple pregnancies [n = 273]). With prolonged WSD feeding, multiple diet switches, and/or increasing age and parity, WSD was associated with increasingly higher insulin levels during glucose tolerance testing, indicative of insulin resistance. The results suggest that prolonged or recurrent calorically dense WSD and/or increased parity, rather than obesity per se, drive excess insulin resistance and metabolic dysfunction.
Supported by ORIP (P51OD011092), NIDDK and NIMH.

 

Early Treatment with a Combination of Two Potent Neutralizing Antibodies Improves Clinical Outcomes and Reduces Virus Replication and Lung Inflammation in SARS CoV-2 Infected Macaques

Van Rompay, Koen K. A. et al., PloS Pathogens, 6 July 2021; Volume 17, Issue 7

https://journals.plos.org/plospathogens/article?id=10.1371/journal.ppat.1009688

The therapeutic efficacy of a combination of two SARS-CoV-2 monoclonal antibodies (mAbs), C135-LS and C144-LS, were investigated in young adult macaques (3 groups of 4 animals; equal sex distribution). Animals were treated intravenously with low or high doses of C135-LS and C144-LS mAbs or control mAb 24 hours post-infection with SARS-CoV-2. Compared to controls, animals treated with either dose of the anti-SARS-CoV-2 mAbs showed improved clinical scores, lower levels of virus replication in upper and lower respiratory tract, and reduced interstitial pneumonia, as measured by lung histology. The study provides proof-of-concept for development of these mAbs for treatment of COVID-19 during early infection.
Supported by ORIP (P51OD011107) and NIAID.

 

Modulation of MHC-E Transport by Viral Decoy Ligands Is Required for RhCMV/SIV Vaccine Efficacy

Verweij, Marieke C. et al., Science. 30 April 2021;372(6541):eabe9233.

https://doi.org/10.1126/science.abe9233

Rhesus cytomegalovirus (RhCMV) strain 68-1-vectored simian immunodeficiency virus (SIV) vaccines elicit strong CD8⁺ T cell responses that can clear SIV infections. Peptides targeted by these T cells are presented on major histocompatibility complex (MHC) II and MHC-E rather than MHC-Ia. Researchers showed that VL9 drives intracellular transport of MHC-E and recognition of RhCMV-infected targets by MHC-E-restricted CD8⁺ T cells. Specific pathogen-free (SPF) rhesus macaques vaccinated with a mutant 68-1 RhCMV lacking VL9 showed no priming of MHC-E-restricted CD8⁺ T cells and no protection against SIV, suggesting that future effective CMV-based HIV vaccines will require MHC-E-restricted CD8⁺ T cell priming.
Supported by ORIP (U42OD023038, P51OD011092), NIAID, NCI, and Bill & Melinda Gates Foundation.

 

In Vitro and in Vivo Functions of SARS-CoV-2 Infection-Enhancing and Neutralizing Antibodies

Li, Dapeng et al., Cell. 18 June 2021;S0092-8674(21)00756-X.

https://doi.org/10.1016/j.cell.2021.06.021

Antibody-dependent enhancement of infection is a concern for clinical use of 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. Cryo-electron microscopy of RBD and NTD antibodies demonstrated function-specific binding modes. RBD and NTD antibodies mediated both neutralization and infection enhancement in vitro. However, infusion of these antibodies into mice or macaques resulted in suppression of virus replication, demonstrating that antibody-enhanced infection in vitro does not necessarily predict enhanced infection in vivo. RBD-neutralizing antibodies having cross-reactivity against coronaviruses were protective against SARS-CoV-2, the most potent of which was DH1047.
Supported by ORIP (P40OD012217, U42OD021458, S10OD018164), NIAID, NCI, NIGMS, NIH Common Fund, Defense Advanced Research Projects Agency, State of North Carolina, New York State, Burroughs Wellcome Fund, Simons Foundation, and Ting Tsung & Wei Fong Chao Foundation

 

A Participant-Derived Xenograft Model of HIV Enables Long-Term Evaluation of Autologous Immunotherapies

McCann, Chase D. et al., Journal of Experimental Medicine. 14 May 2021;218(7):e20201908.

https://doi.org/10.1084/jem.20201908

HIV-specific CD8⁺ T cells partially control viral replication but rarely provide lasting protection due to immune escape. Investigators showed that engrafting NSG mice with memory CD4⁺ T cells from HIV⁺ donors enables evaluation of autologous T cell responses while avoiding graft-versus-host disease. Treating HIV-infected mice with clinically relevant T cell products reduced viremia. In vivo activity was significantly enhanced when T cells were engineered with surface-conjugated nanogels carrying an Interleukin-15 superagonist but was ultimately limited by the pervasive selection of escape mutations, recapitulating human patterns. This “participant-derived xenograft” model provides a powerful tool for developing T cell-based therapies for HIV.
Supported by ORIP (R01OD011095), NIAID, NIDA, NIMH, NINDS, NCATS, Canadian Institutes of Health Research, and Michael Smith Foundation for Health Research

 

Mitigation of Endemic GI-Tract Pathogen-Mediated Inflammation Through Development of Multimodal Treatment Regimen and Its Impact on SIV Acquisition in Rhesus Macaques

Bochart, Rachele M. et al., PLoS Pathogens. 10 May 2021;17(5):e1009565.

https://doi.org/10.1371/journal.ppat.1009565

In addition to being premier HIV models, rhesus macaques are models for other infectious diseases and colitis, where background colon health and inflammation may confound results. Starting with the standard SPF model, researchers established a gastrointestinal pathogen-free (GPF) colony via multimodal therapy (enrofloxacin, azithromycin, fenbendazole, and paromomycin) to eliminate common endemic pathogens (EPs). This treatment combined with continued pathogen exclusion eliminated common EPs, improved mucosal barriers, and reduced mucosal and systemic inflammation without microbiota disruption. GPF animals challenged with SIV intrarectally demonstrated a more controlled and consistent rate of SIV acquisition, suggesting the value of this model for HIV studies.
Supported by ORIP (U42OD023038, P51OD011092), NCI, and NIAID.

 

Protection of Newborn Macaques by Plant-Derived HIV Broadly Neutralizing Antibodies: A Model for Passive Immunotherapy During Breastfeeding

Rosenberg, Yvonne J. et al., Journal of Virology. 30 June 2021;JVI0026821; Online ahead of print.

https://doi.org/10.1128/JVI.00268-21

Preventing vertical transmission of HIV to newborns is an unmet medical need in resource poor countries. Using a breastfeeding macaque model with multiple simian-human immunodeficiency virus challenge, researchers assessed the protective efficacy of two human broadly neutralizing antibodies (bnAbs) against HIV, PGT121 and VRC07-523, which are produced by a plant expression system. Despite the transient presence of plasma viral RNA, the bnAbs prevented productive infection in all newborns with no sustained plasma viremia, compared to viral loads ranging from 10³ to 5x10⁸ in four untreated controls. Thus, plant-expressed antibodies show promise as passive immunoprophylaxis in a breastfeeding model in newborns.
Supported by ORIP (U42OD023038, P51OD011092) and NIAID.

 

Recrudescence of Natural Coccidioidomycosis During Combination Antiretroviral Therapy in a Pigtail Macaque Experimentally Infected with Simian Immunodeficiency Virus

Guerriero, Kathryn A. et al., AIDS Research and Human Retroviruses. July 2021;37(7):505-509.

https://doi.org/10.1089/AID.2020.0228

Coccidioidomycosis is a common fungal infection in people living with HIV, particularly in regions where Coccidioides is endemic, such as the US Southwest. Researchers diagnosed a recrudescent case of previously treated, naturally occurring coccidioidomycosis in a pigtail macaque experimentally infected with simian immunodeficiency virus (SIV) and virally suppressed on combination antiretroviral therapy (cART). Coccidioides IgG antibody titer became detectable before discontinuation of cART, but symptomatic coccidioidomycosis developed after cART withdrawal. This animal was screened and treated in accordance with the guidelines for coccidioidomycosis prevention and treatment. The researchers conclude that macaques with coccidioidomycosis history should be excluded from HIV studies.
Supported by ORIP (P51OD010425), NIAID, and NIMH.

 

Cell-specific Transcriptional Control of Mitochondrial Metabolism by TIF1γ Drives Erythropoiesis

Rossmann, Marlies P. et al. Science. 14 May 2021;372(6543):716-721. PMC8177078

https://science.sciencemag.org/content/372/6543/716

Transcription and metabolism both influence cell function but dedicated transcriptional control of metabolic pathways that regulate cell fate has rarely been defined. The authors discovered that inhibition of the pyrimidine biosynthesis enzyme dihydroorotate dehydrogenase (DHODH) rescues erythroid differentiation in bloodless zebrafish moonshine (mon) mutant embryos defective for transcriptional intermediary factor 1 gamma (tif1γ). Upon tif1γ loss, CoQ levels are reduced, and a high succinate/α-ketoglutarate ratio leads to increased histone methylation. A CoQ analog rescues mon's bloodless phenotype. These results demonstrate that mitochondrial metabolism is a key output of a lineage transcription factor that drives cell fate decisions in the early blood lineage.
Supported by ORIP (R24 OD017870), NIGMS, NHLBI, NCI.

 

Identification of Basp1 as a Novel Angiogenesis-regulating Gene by Multi-model System Studies

Khajavi, Mehrdad et al. Federation of American Societies for Experimental Biology (FASEB). 2021 May;35(5):e21404. PMC8218237

https://pubmed.ncbi.nlm.nih.gov/33899275/

The authors previously used genetic diversity in inbred mouse strains to identify quantitative trait loci (QTLs) responsible for differences in angiogenic response. Employing a mouse genome-wide association study (GWAS) approach, the region on chromosome 15 containing Basp1 was identified as being significantly associated with angiogenesis in inbred strains. To investigate its role in vivo, they knocked out basp1 in transgenic kdrl:zsGreen zebrafish embryos using a widely adopted CRISPR-Cas9 system. They further showed that basp1 promotes angiogenesis by upregulating β-catenin gene and the Dll4/Notch1 signaling pathway. These results provide the first in vivo evidence to indicate the role of Basp1 as an angiogenesis-regulating gene.
Supported by ORIP (R24 OD017870), NEI, HHMI.

 

Acoustofluidic Rotational Tweezing Enables High-speed Contactless Morphological Phenotyping of Zebrafish Larvae

Chen, Chuyi et al. Nature Communications 18 February 2021;12(1):1118. PMC7892888

https://www.nature.com/articles/s41467-021-21373-3

These authors demonstrate an acoustofluidic rotational tweezing platform that enables contactless, high-speed, 3D multispectral imaging and digital reconstruction of zebrafish larvae for quantitative phenotypic analysis. The acoustic-induced polarized vortex streaming achieves contactless and rapid (~1 s/rotation) rotation of zebrafish larvae enabling multispectral imaging of the zebrafish body and internal organs. They developed a 3D reconstruction pipeline that yields accurate 3D models based on the multi-view images for quantitative evaluation. With its contactless nature and advantages in speed and automation, the acoustofluidic rotational tweezing system has the potential to be a valuable asset for developmental biology and pre-clinical drug development in pharmacology.
Supported by ORIP (R43 OD024963), NCI, NIGMS.

 

Fluorescence-based Sorting of Caenorhabditis Elegans via Acoustofluidics

Zhang, Jinxin et al. Lab on a Chip. 19 May 2020(10):1729-1739. PMC7239761.

https://pubs.rsc.org/en/content/articlelanding/2020/lc/d0lc00051

The authors present an integrated acoustofluidic chip capable of identifying worms of interest based on expression of a fluorescent protein in a continuous flow and then separate them in a high-throughput manner. Utilizing planar fiber optics, their acoustofluidic device requires no temporary immobilization of worms for interrogation/detection, thereby improving the throughput. The device can sort worms of different developmental stages (L3 and L4 stage worms) at high throughput and accuracy. In their acoustofluidic chip, the time to complete the detection and sorting of one worm is only 50 ms, which outperforms nearly all existing microfluidics-based worm sorting devices.
Supported by ORIP (R43 OD024963), NIEHS, NIDDK.

 

Larval Zebrafish Use Olfactory Detection of Sodium and Chloride to Avoid Salt Water

Herrera, Kristian J. et al. Current Biology. 22 February 2021;31(4):782-793.e3. PMC7904661.

https://www.sciencedirect.com/science/article/pii/S0960982220317644

Zebrafish are freshwater fish unable to tolerate high-salt environments and would benefit from neural mechanisms that enable the navigation of salt gradients to avoid high salinity. Yet zebrafish lack epithelial sodium channels, the primary conduit land animals use to taste sodium. This suggests fish may possess novel, undescribed mechanisms for salt detection. In the present study, the authors show that zebrafish indeed respond to small temporal increases in salt by reorienting more frequently. In summary, this study establishes that zebrafish larvae can navigate and thus detect salinity gradients and that this is achieved through previously undescribed sensory mechanisms for salt detection.
Supported by ORIP (R43 OD024879, R44 OD024879), NINDS.

 

Algorithms Underlying Flexible Phototaxis in Larval Zebrafish

Chen, Alex B. et al. Journal of Experimental Biology. 24 May 2021;224(10):jeb238386. PMC8180250.

https://journals.biologists.com/jeb/article/224/10/jeb238386/268333/Algorithms-underlying-flexible-phototaxis-in

Given that physiological and environmental variables undergo constant fluctuations over time, how do biological control systems maintain control over these values? The authors demonstrate that larval zebrafish use phototaxis to maintain environmental luminance at a set point, that the value of this set point fluctuates on a time scale of seconds when environmental luminance changes, and it is determined by calculating the mean input across both sides of the visual field. Feedback from the surroundings drives allostatic changes to the luminance set point. The authors describe a novel behavioral algorithm with which larval zebrafish exert control over a sensory variable.
Supported by ORIP (R43 OD024879, R44 OD024879), NINDS.

 

A Platform for Experimental Precision Medicine: The Extended BXD Mouse Family

Ashbrook, David G. et al., Cell Systems, 17 March 2021; Volume 12, Issue 3, Pages 235-247

https://www.sciencedirect.com/science/article/abs/pii/S2405471220305032

Systems genetics using rodent models has recently been revitalized thanks to several resources:   the BXD family, the Hybrid Mouse Diversity Panel, and the Collaborative Cross. The main limitation has been modest mapping power and precision due to small strain numbers. Investigators expanded the BXD family of mice to 140 fully isogenic strains. Heritable traits can be mapped with precision. Current BXD phenomes include much omics data and thousands of quantitative traits. BXDs can be extended by a single-generation cross up to 19,460 isogenic F1 progeny.  This extended BXD family is an effective platform for testing causal modeling and predictive validation.
The BXD Resource at the Jackson Laboratory is supported by ORIP P40 OD011102 awarded to Dr. Cathleen M. Lutz.

 

A Novel Non-human Primate Model of Pelizaeus-Merzbacher Disease (PMD)

Sherman, Larry S. et al., ScienceDirect, October 2021; Volume 158

https://www.sciencedirect.com/science/article/pii/S096999612100214X

Pelizaeus-Merzbacher (PMD) disease in humans is a severe hypomyelinating disorder of the central nervous system (CNS) linked to mutations in the proteolipid protein-1 (PLP1) gene. Investigators report on three spontaneous cases of male neonatal rhesus macaques (RMs) with clinical symptoms of hypomyelinating disease. Genetic analysis revealed that the parents of these related RMs carried a rare, hemizygous missense variant in exon 5 of the PLP1 gene. These RMs represent the first reported NHP model of PMD, providing an opportunity for studies to promote myelination in pediatric hypomyelinating diseases as other animal models of PMD do not fully mimic human disorders.
Supported by ORIP (R24OD021324, P51OD011092, and S10OD025002) and NINDS.

 

'Enhancing' Red Cell Fate through Epigenetic Mechanisms

Rossmann, Marlies P. and Zon, Leonard I., Current Opinion in Hematology, 1 May 2021; Volume 28, Issue 3, Pages 129-137

https://journals.lww.com/co-hematology/Fulltext/2021/05000/_Enhancing__red_cell_fate_through_epigenetic.2.aspx

Transcription of erythroid-specific genes is regulated by the three-dimensional (3D) structure and composition of chromatin, which changes during erythroid differentiation. These authors address recent developments delineating the interface of chromatin regulation and erythroid-specific lineage transcription. They survey the erythroid chromatin landscape, erythroid enhancer-promotor interactions, super-enhancer functionality, the role of chromatin modifiers and epigenetic crosstalk, as well as the progress in mapping red blood cell (RBC) trait-associated genetic variants within cis-regulatory elements (CREs) identified in genome-wide association study (GWAS) efforts. New emerging technologies allow investigation of small cell numbers have advanced our understanding of chromatin dynamics during erythroid differentiation in vivo.
Supported by ORIP (R24 OD017870), NHLBI.

 

Neutralizing Antibody Vaccine for Pandemic and Pre-emergent Coronaviruses

Saunders, Kevin O. et al., Nature, 2021 June, Volume 594, Pages 553-559

https://www.nature.com/articles/s41586-021-03594-0#citeas

Betacoronaviruses caused outbreaks of severe acute respiratory syndrome (SARS) and Middle East respiratory syndrome, as well as the pandemic of SARS coronavirus 2 (SARS-CoV-2)1-4.  Here the authors show that the immunization of macaques with nanoparticles conjugated with the receptor-binding domain of SARS-CoV-2, and adjuvanted with 3M-052 and alum, elicits cross-neutralizing antibody responses against bat coronaviruses, SARS-CoV, and SARS-CoV-2 (including the B.1.1.7, P.1 and B.1.351 variants). Nucleoside-modified mRNAs that encode a stabilized transmembrane spike or monomeric receptor-binding domain also induced cross-neutralizing antibody responses against SARS-CoV-2. These results demonstrate that current mRNA-based vaccines may provide some protection from future outbreaks of zoonotic betacoronaviruses.
Supported by ORIP (P40 OD012217, U42 OD021458), NIAID, NCI.

 

Loss of Gap Junction Delta-2 (GJD2) Gene Orthologs Leads to Refractive Error in Zebrafish

Quint, Wim H. et al., Communications Biology, 2021 June 3, Volume 4, Pages 1-14

https://www.nature.com/articles/s42003-021-02185-z

Myopia is the most common developmental disorder of juvenile eyes. Although little is known about the functional role of GJD2 in refractive error development, the authors find that depletion of gjd2a (Cx35.5) or gjd2b (Cx35.1) orthologs in zebrafish cause changes in eye biometry and refractive status. Their immunohistological and scRNA sequencing studies show that Cx35.5 (gjd2a) is a retinal connexin; its depletion leads to hyperopia and electrophysiological retina changes. They found a lenticular role; lack of Cx35.1 (gjd2b) led to a nuclear cataract that triggered axial elongation. The results provide functional evidence of a link between gjd2 and refractive error.
Supported by ORIP (R24 OD026591), NIGMS, NINDS.

 

Interneuron Origins in the Embryonic Porcine Medial Ganglionic Eminence

Casalia, Mariana L. et al, Journal of Neuroscience, 7 April 2021, Volume 41, Issue 14, Pages 3105-3119

https://www.jneurosci.org/content/41/14/3105

The authors report that transcription factor expression patterns in porcine embryonic subpallium are similar to rodents. Their findings reveal that porcine embryonic MGE progenitors could serve as a valuable source for interneuron-based xenotransplantation therapies. They demonstrate that porcine medial ganglionic eminence exhibits a distinct transcriptional and interneuron-specific antibody profile, in vitro migratory capacity, and are amenable to xenotransplantation. This is the first comprehensive examination of embryonic interneuron origins in the pig; because a rich neurodevelopmental literature on embryonic mouse medial ganglionic eminence exists (with some additional characterizations in monkeys and humans), their work allows direct neurodevelopmental comparisons with this literature.
Supported by ORIP (U42 OD011140), NINDS.

 

Mineralocorticoid Receptor Blockade Normalizes Coronary Resistance in Obese Swine Independent of Functional Alterations in Kv Channels

Goodwill, Adam G. et al., Basic Research in Cardiology, 20 May 2021; Volume 116, Article 35

https://link.springer.com/article/10.1007/s00395-021-00879-3#citeas

Impaired coronary microvascular function (e.g., reduced dilation and coronary flow reserve) predicts cardiac mortality in obesity. Mineralocorticoid receptor (MR) antagonism improves coronary microvascular function in obese humans and animals. Inhibition of Kv channels reduced coronary blood flow and augmented coronary resistance under baseline conditions in lean but not obese swine and had no impact on hypoxemic coronary vasodilation. MR blockade prevented obesity-associated coronary arteriolar stiffening independent of cardiac capillary density and changes in cardiac function. These data indicate that chronic MR inhibition prevents increased coronary resistance in obesity independent of Kv channel function and is associated with mitigation of obesity-mediated coronary arteriolar stiffening.
Supported by ORIP (U42 OD011140, S10 OD023438), NHLBI, NIBIB.

 

A Symphony of Destruction: Dynamic Differential Fibrinogenolytic Toxicity by Rattlesnake (Crotalus and Sistrurus) Venoms

Seneci, Lorenzo et al., Comparative Biochemistry and Physiology Part C: Toxicology and Pharmacology, 2021 July, Volume 245, 109034

https://www.sciencedirect.com/science/article/pii/S1532045621000612

This study adopts rattlesnakes as a model group to investigate the evolutionary history of venom coagulotoxicity in the context of phylogenetics, natural history, and biology. Venom-induced clotting of human plasma and fibrinogen was determined and mapped onto the rattlesnake phylogenetic tree to reconstruct the evolution of coagulotoxicity across the group. Results indicate that venom phenotype is often independent of phylogenetic relationships in rattlesnakes, suggesting the importance of diet and/or other environmental variables. This study is the most comprehensive effort to date to characterize the evolutionary and biological aspects of coagulotoxins in rattlesnake venom. Further research at finer taxonomic levels is recommended.
Supported by ORIP (P40 OD010960).

 

Characterization of Axolotl Lampbrush Chromosomes by Fluorescence in Situ Hybridization and Immunostaining

Keinath, Melissa C., et al., Experimental Cell Research, 15 April 2021;401(2):112523

https://pubmed.ncbi.nlm.nih.gov/33675804/

The lampbrush chromosomes (LBCs) in oocytes of the Mexican axolotl (Ambystoma mexicanum) were identified by their relative lengths and predicted centromeres; they have never been associated completely with the mitotic karyotype, linkage maps, or genome assembly. The authors identified nine of the axolotl LBCs using RNAseq to identify actively transcribed genes and thirteen BAC (bacterial artificial clone) probes containing pieces of active genes. This study presents a simple and reliable way to identify each axolotl LBC cytologically and to anchor chromosome-length sequences to the LBCs by immunostaining and fluorescence in situ hybridization. This data will facilitate a more detailed analysis of LBC loops.
Supported by ORIP (P40 OD019794, R24 OD010435), NIGMS.

 

Advancing Human Disease Research with Fish Evolutionary Mutant Models

Beck, Emily A., et al., Trends in Genetics, 29 July 2021;S0168-9525(21)00191-8 (article acknowledge ORIP workshop session “Validation of Non-Zebrafish Aquatic Models for Preclinical Research”)

https://www.cell.com/trends/genetics/fulltext/S0168-9525(21)00191-8

Model organism research is essential to understand disease mechanisms. However, laboratory-induced genetic models can lack genetic variation and often fail to mimic disease severity. Evolutionary mutant models (EMMs) are species with evolved phenotypes that mimic human disease. They have improved our understanding of cancer, diabetes, and aging. Fish are the most diverse group of vertebrates, exhibiting a kaleidoscope of specialized phenotypes, many that would be pathogenic in humans but are adaptive in the species' specialized habitat. Evolved compensations can suggest avenues for novel disease therapies. This review summarizes current research using fish EMMs to advance our understanding of human disease.
Supported by ORIP (R01 OD011116), NIA, NIDA, NIGMS, NSF.

 

Factor XII Plays a Pathogenic Role in Organ Failure and Death in Baboons Challenged with Staphylococcus aureus

Silasi, Robert et al., Blood, 15 July 2021;138(2):178-189

(Article was featured on the journal’s cover: https://ashpublications.org/blood/issue/138/2).

(Article’s Editorial commentary: https://ashpublications.org/blood/article/138/2/107/476358/FXII-inhibition-multipronged-benefits)

Activation of coagulation factor (F) XI promotes multiorgan failure in rodent models of sepsis and in a baboon model of lethal systemic inflammation induced by infusion of heat-inactivated Staphylococcus aureus. The authors used the anticoagulant FXII-neutralizing antibody 5C12 to verify the mechanistic role of FXII. Inhibition of FXII prevented fever, terminal hypotension, respiratory distress, and multiorgan failure. All animals receiving 5C12 had milder and transient clinical symptoms; untreated control animals suffered irreversible multiorgan failure. This study confirms their previous finding that at least two enzymes of FXIa and FXIIa play critical roles in the development of an acute and terminal inflammatory response.
Supported by ORIP (P40 OD024628), NIAID, NHLBI, NIGMS.

 

Systems Vaccinology of the BNT162b2 mRNA Vaccine in Humans

Arunachalam, Prabhu S. et al., Nature, 2021 Jul 12;596(7872):410-416

https://doi.org/10.1038/s41586-021-03791-x

It was poorly understood how mRNA vaccines against SARS-CoV-2 stimulate protective immune responses. To address this, researchers comprehensively profiled innate and adaptive immune responses of healthy volunteers vaccinated with the Pfizer-BioNTech mRNA vaccine (BNT162b2). Vaccination resulted in robust production of neutralizing antibodies against wild-type SARS-CoV-2, to a lesser extent, the beta variant, as well as significant increases in antigen-specific polyfunctional CD4⁺ and CD8⁺ T cells after the second dose. Booster vaccination stimulated an enhanced innate immune response compared to primary vaccination, demonstrating the capacity of BNT162b2 to prime the innate immune system to mount a more potent response after booster immunization.
Supported by ORIP (P51OD 011132, S10OD026799), NIAID, Open Philanthropy, Sean Parker Cancer Institute, Soffer endowment, Violetta Horton endowment, Stanford University, Henry Gustav Floren Trust, Parker Foundation, and Crown Foundation.

 

Modified Adenovirus Prime–Protein Boost Clade C HIV Vaccine Strategy Results in Reduced Viral DNA in Blood and Tissues Following Tier 2 SHIV Challenge

Malherbe, Delphine C. et al., Frontiers in Immunology, 2021 Feb 15;11:626464

https://doi.org/10.3389/fimmu.2020.626464

Researchers conducted a comparative vaccine challenge study in rhesus macaques. One group of monkeys was vaccinated using co-immunization with DNA Gag and Env expression plasmids and trimeric Env gp140 glycoprotein. The other group was primed with two replicating simian adenovirus-vectored vaccines expressing Gag and boosted with trimeric Env gp140. Both strategies elicited antigen-specific humoral and cellular immune responses, but neither approach provided significant protection from viral acquisition upon repeated mucosal challenges with a heterologous Tier 2 SHIV. Nevertheless, both regimens significantly lowered cell-associated viral DNA in multiple tissues, thus potentially dampening the infection and providing clues for further vaccine development.
Supported by ORIP (U42OD023038, P51OD011092) and NIAID.

 

Interleukin-15 Response Signature Predicts RhCMV/SIV Vaccine Efficacy

Barrenäs, Fredrik et al., PLoS (Public Library of Science) Pathogens, 2021 Jul 6;17(7):e1009278

https://doi.org/10.1371/journal.ppat.1009278

Standard immunogenicity measures do not predict efficacy of a vaccine based on strain 68-1 rhesus cytomegalovirus (RhCMV) vectors expressing SIV proteins (RhCMV/SIV). This vaccine robustly protects just over half of immunized monkeys. Using functional genomics, researchers found that RhCMV/SIV efficacy is correlated with a vaccine-induced response to interleukin-15 (IL-15) that includes modulation of immune cell, inflammation, toll-like receptor signaling, and cell death programming pathways. RhCMV/SIV imparts a coordinated and persistent induction of innate and adaptive immune pathways featuring IL-15, a known regulator of CD8⁺ T cell function, that support the ability of vaccine-elicited CD8⁺ T cells to mediate protection against SIV.
Supported by ORIP (P51OD010425, P51OD011092), NIAID, and NCI.

 

Combining In Vivo Corneal Confocal Microscopy With Deep Learning-Based Analysis Reveals Sensory Nerve Fiber Loss in Acute Simian Immunodeficiency Virus Infection

McCarron, Megan E. et al., Cornea, 2021 May 1;40(5):635-642

https://doi.org/10.1097/ICO.0000000000002661

Researchers characterized corneal subbasal nerve plexus features of normal and simian immunodeficiency virus (SIV)-infected pigtail and rhesus macaques using in vivo confocal microscopy and a deep learning approach for automated assessments. Corneal nerve fiber length and fractal dimension measurements did not differ between species, but pigtail macaques had significantly higher baseline corneal nerve fiber tortuosity than rhesus macaques. Acute SIV infection induced decreased corneal nerve fiber length and fractal dimension in the pigtail macaque model of HIV. Adapting deep learning analyses to clinical corneal nerve assessments will improve monitoring of small sensory nerve fiber damage in numerous clinical contexts, including HIV.
Supported by ORIP (U42OD013117), NINDS, Johns Hopkins University School of Medicine, and Fulbright New Zealand.

 

Single-cell Protein Activity Analysis Identifies Recurrence-associated Renal Tumor Macrophages

Obradovic, Aleksandar et al., Cell, 27 May 2021;184(11),P2988-3005.e16.

https://doi.org/10.1016/j.cell.2021.04.038

Post-surgery course of clear cell renal carcinoma (ccRCC) is mixed because of the heterogeneity of the disease. Using high-performance computing cluster and storage systems funded by the NIH Shared Instrumentation Programs, investigators established an inclusive ccRCC tumor microenvironment (TME) map by using single-cell RNA sequencing data of subpopulations of tumor and tumor-adjacent tissues. Analysis of the data identified key TME subpopulations as well as their master regulators and candidate cell-cell interactions, revealing clinically relevant cell populations. Specifically, the study uncovered a tumor-specific macrophage subpopulation, validated by spatially resolved, quantitative multispectral immunofluorescence. In a large clinical validation cohort, markers of this subpopulation were significantly enriched in tumors from patients who recurred following surgery.
Supported by ORIP (S10 OD012351, S10 OD021764).

 

Thioesterase Superfamily Member 1 Undergoes Stimulus-coupled Conformational Reorganization to Regulate Metabolism in Mice

Li, Yue et al., Nature Communications, 9 June 2021;12(3493).

https://doi.org/10.1038/s41467-021-23595-x

Thermogenesis is suppressed in brown adipose tissue by thioesterase superfamily member 1 (Them1), a long chain fatty acyl-CoA thioesterase. Them1 is highly upregulated by cold ambient temperature, where it reduces fatty acid availability and limits thermogenesis. Investigators show that Them1 regulates metabolism by undergoing conformational changes in response to β-adrenergic stimulation that alter Them1 intracellular distribution. Them1 forms metabolically active puncta near lipid droplets and mitochondria. Upon stimulation, Them1 is phosphorylated at the N-terminus, inhibiting puncta formation and activity, and resulting in a diffuse intracellular localization. Investigators show that Them1 puncta are biomolecular condensates that are inhibited by phosphorylation. Them1 forms intracellular biomolecular condensates that limit fatty acid oxidation and suppress thermogenesis. When energy is demanded, the condensates are disrupted by phosphorylation to allow for maximal thermogenesis. The stimulus-coupled reorganization of Them1 provides fine-tuning of thermogenesis and energy expenditure.
Supported by ORIP (S10 OD019988).

 

Phase Separation Drives Aberrant Chromatin Looping and Cancer Development

Ahn, Jeong Hyun et al., Nature, 2021 Jul;595(7868):591-595.

https://doi.org/10.1038/s41586-021-03662-5

How unstructured intrinsically disordered regions (IDRs) contribute to oncogenesis is elusive. Using an ORIP-supported Orbitrap Fusion Tribrid Mass Spectrometer, investigators show that IDRs contained within NUP98–HOXA9, a homeodomain-containing transcription factor chimera recurrently detected in leukaemias, are essential for establishing liquid–liquid phase separation (LLPS) puncta of chimera and for inducing leukaemic transformation. LLPS of NUP98–HOXA9 not only promotes chromatin occupancy of chimera transcription factors, but also is required for the formation of a broad “super-enhancer”-like binding pattern typically seen at leukaemogenic genes, which potentiates transcriptional activation. An artificial HOX chimera, created by replacing the phenylalanine and glycine repeats of NUP98 with an unrelated LLPS-forming IDR of the FUS protein, had similar enhancing effects on the genome-wide binding and target gene activation of the chimera. This report describes a proof-of-principle example in which cancer acquires mutation to establish oncogenic transcription factor condensates via phase separation, which simultaneously enhances their genomic targeting and induces organization of aberrant three-dimensional chromatin structure during tumor transformation.
Supported by ORIP (S10 OD018445).

 

Cryopreservation Method for Drosophila Melanogaster Embryos

Zhan, Li. et al., Nature Communications, 2021 April 23;12(1):2412

https://www.nature.com/articles/s41467-021-22694-z

D. melanogaster is a premier model for biomedical research. However, preservation of Drosophila stocks is labor intensive and costly to pass live cultures monthly to fresh food. Researchers at University of Minnesota reported an efficient method for cryopreservation by optimizing key steps including embryo permeabilization and cryoprotectant agent loading. This method resulted in >10% of embryos developing into fertile adults after cryopreservation for 25 distinct strains from different sources.  The further optimization and wide adoption of this protocol will solve the long-standing issue in reliably preserving Drosophila stocks and will significantly impact Drosophila as a model organism for biomedical research.

Supported by ORIP (1R21OD028758), NIGMS, NSF, and other sources.

 

MRI Characteristics of Japanese Macaque Encephalomyelitis (JME): Comparison to Human Diseases

Tagge, Ian J. et al., 2021; Journal of Neuroimaging, 2021 May;31(3):480-492

https://onlinelibrary.wiley.com/doi/10.1111/jon.12868

MRI data were obtained from 114 Japanese macaques including 30 animals of both sexes that presented with neurological signs of JME. Quantitative estimates of blood-brain-barrier permeability to gadolinium-based-contrast agent (GBCA) were obtained in acute, GBCA-enhancing lesions, and longitudinal imaging data were acquired for 15 JME animals. Intense, focal neuroinflammation was a key MRI finding in JME. Several features of JME compare directly to human inflammatory demyelinating diseases. The development and validation of noninvasive imaging biomarkers in JME provides the potential to improve diagnostic specificity and contribute to the understanding of human demyelinating diseases.

Supported by ORIP (P51OD011092, S10OD018224), NINDS, and NIBIB.

 

Evidence in Primates Supporting the Use of Chemogenetics for the Treatment of Human Refractory Neuropsychiatric Disorders

Roseboom, Patrick H. et al., Molecular Therapy, 2021 April 23;S1525-0016(21)00209-4

https://doi.org/10.1016/j.ymthe.2021.04.021

A rhesus macaque model of pathological anxiety was used to investigate the feasibility of decreasing anxiety using chemogenetics, known as DREADDs (designer receptors exclusively activated by designer drugs), to reduce amygdala neuronal activity. A low-dose clozapine administration strategy was developed to induce DREADD-mediated amygdala inhibition. Compared to controls, clozapine selectively decreased anxiety-related freezing behavior in the human intruder paradigm in the chemogentic monkeys, while coo vocalizations and locomotion were unaffected. These results are an important step in establishing chemogenetic strategies for patients with refractory neuropsychiatric disorders in which amygdala alterations are central to disease pathophysiology.

Supported by ORIP (P51OD011106), NIMH, and NICHD. 

 

Rhesus Macaques Build New Social Connections After a Natural Disaster

Testard, Camille et al. Current Biology. 2021 April 8; S0960-9822(21)00368-7

https://www.sciencedirect.com/science/article/pii/S0960982221003687

Climate change has increased the frequency and intensity of weather-related disasters such as hurricanes and floods. In 2017, Puerto Rico suffered its worst natural disaster, Hurricane Maria, leaving 3,000 dead and provoking a mental health crisis. Cayo Santiago Island, home to a population of rhesus macaques (Macaca mulatta), was devastated by this storm. Testard et al. compared social networks of two groups of macaques before and after the hurricane and found an increase in affiliative social connections, driven largely by monkeys most socially isolated before Hurricane Maria. Further analysis revealed monkeys invested in building new relationships rather than strengthening existing ones.

Supported by ORIP (P40 OD012217), NIA, NIMH

 

Functional Convergence of a Germline-Encoded Neutralizing Antibody Response in Rhesus Macaques Immunized with HCV Envelope Glycoproteins

Chen, Fang et al., Immunity. 2021 Apr 13; 54(4):781-796

https://doi.org/10.1016/j.immuni.2021.02.013

Immunoglobulin heavy chain variable gene IGHV1-69-encoded broadly neutralizing antibodies (bnAbs) targeting the hepatitis C virus (HCV) envelope glycoprotein (Env) E2 are important for protection against HCV infection in humans. An IGHV1-69 ortholog, VH1.36, is preferentially used for bnAbs isolated from rhesus macaques immunized against HCV Env. Researchers investigated the genetic, structural, and functional properties of VH1.36-encoded bnAbs generated by HCV Env vaccination of macaques and compared their findings to IGHV1-69-encoded bnAbs from HCV patients. The investigators found that macaque VH1.36- and human IGHV1-69-encoded bnAbs share many common features, which provides an excellent framework for rational HCV vaccine design and testing.

Supported by ORIP (P51OD011133, U42OD010442), NIAID, NCI, NIGMS, Dept. of Energy, Independent Research Fund Denmark, Novo Nordisk Foundation, Candys Foundation, and Lundbeck Foundation

 

Psychosocial Stress Alters the Immune Response and Results in Higher Viral Load During Acute SIV Infection in a Pigtailed Macaque Model of HIV

Guerrero-Martin, Selena et al., Journal of Infectious Diseases. 2021 May 10; Online ahead of print.

https://doi.org/10.1093/infdis/jiab252

Social distancing is an important countermeasure for a pandemic, but social isolation may also have adverse health outcomes in the context of infectious diseases, such as HIV. Researchers compared commonly measured parameters of HIV progression between singly and socially housed simian immunodeficiency virus (SIV)-infected pigtailed macaques. Throughout acute SIV infection, singly housed pigtailed macaques had a higher viral load in the plasma and cerebrospinal fluid and demonstrated greater CD4⁺ T cell declines and more CD4⁺ and CD8⁺ T cell activation compared to socially housed macaques. These findings suggest that psychosocial stress could augment the progression of HIV infection.

Supported by ORIP (U42OD013117, P40OD013117, K01OD018244), NIAID, NINDS, NIMH, AALAS Grants for Laboratory Animal Science, and Blaustein Pain Foundation.

 

Creb5 Establishes the Competence for Prg4 Expression in Articular Cartilage

Zhang, Cheng-Hai et al., Communications Biology. 2021 Mar 12; 4(1):332

https://doi.org/10.1038/s42003-021-01857-0

Cells comprising the superficial zone of articular cartilage express lubricin, encoded by the Prg4 gene, that lubricates joints. Researchers identified Creb5 as a transcription factor that is required for TGF-β and EGFR signaling to induce Prg4 expression. Forced expression of Creb5 in deep-zone chondrocytes of articular cartilage confers competence for TGF-β and EGFR signals to induce Prg4 expression. The researchers showed that Creb5 directly binds to two Prg4 promoter-proximal regulatory elements, which work together with a more distal regulatory element to drive induction of Prg4 by TGF-β. Thus, Creb5 is a critical regulator of Prg4/lubricin expression in the articular cartilage.

Supported by ORIP (U42OD11158), NIAMS, NIDDK, and Arthritis National Research Foundation.

 

Nonhuman Primate Models for SARS-CoV-2 Research: Cryopreservation as a Means to Maintain Critical Models and Enhance the Genetic Diversity of Colonies

Arnegard, Matthew and Hild, Sheri. 2021 May 24. Online ahead of print.

https://doi.org/10.1038/s41684-021-00792-1

This commentary, written by ORIP staff, addresses the need for improved cryopreservation methods and resources for nonhuman primate (NHP) gametes and embryos to safeguard newly developed NHP models and enhance the genetic diversity of NHP colonies without reliance on animal importations. Cryopreservation also plays critical roles in medical approaches to preserve the fertility of patients who must undergo potentially gonadotoxic treatments, as well as nascent genome editing efforts to develop new NHP models for human diseases. Given these diverse benefits to research progress, ORIP continues to fund the development of cryopreservation tools and approaches for NHPs and other animal models.

 

A Chromosome-level Genome of Astyanax Mexicanus Surface Fish for Comparing Population-specific Genetic Differences Contributing to Trait Evolution

Warren, Wesley C. et al. Nature Communications 2021 Mar 4;12(1):1447 (PMC7933363)

https://www.nature.com/articles/s41467-021-21733-z

Surface-dwelling populations of the cave-dwelling Astyanax mexicanus are well adapted to subterranean life. Warren et al. present a chromosome-level surface fish genome and performed quantitative trait locus (QTL) mapping analyses finding new candidate genes for eye loss such as dusp26. They used CRISPR gene editing in A. mexicanus to confirm the essential role of a gene within an eye size QTL, rx3, in eye formation; they generated the first genome-wide evaluation of deletion variability across cavefish populations. This surface fish genome reference provides a more complete resource for comparative, functional and genetic studies of drastic trait differences within a species.

Supported by ORIP (R24 OD011198), NIA, NICHD, NIGMS, NIDCR.

 

The SARS-CoV-2 Receptor and Other Key Components of the Renin-Angiotensin-Aldosterone System Related to COVID-19 are Expressed in Enterocytes in Larval Zebrafish

Postlethwait, John H. et al. Biology Open (2021) 10, bio058172. doi:10.1242/bio.058172

https://bio.biologists.org/content/10/3/bio058172.article-info

Hypertension and respiratory inflammation are exacerbated by the Renin-Angiotensin-Aldosterone System (RAAS), which normally protects from dropping blood pressure via Angiotensin II (Ang II) produced by the enzyme Ace. The Ace paralog Ace2 degrades Ang II and serves as the SARS-CoV-2 receptor. To exploit zebrafish to understand the relationship of RAAS to COVID-19, the group conducted genomic and phylogenetic analyses. Results identified a type of enterocyte as the expression site of zebrafish orthologs of key RAAS components, including the SARSCoV-2 co-receptor.  Results identified vascular cell subtypes expressing Ang II receptors and identified cell types to exploit zebrafish as a model for understanding COVID-19 mechanisms.

Supported by ORIP (R24 OD026591, R01 OD011116), NIGMS, NICHD.

 

A Novel Tau-Based Rhesus Monkey Model of Alzheimer’s Pathogenesis

Beckman, Danielle et al., Alzheimer’s & Dementia. 2021 Mar 18; Epub ahead of print.

https://pubmed.ncbi.nlm.nih.gov/33734581/

Alzheimer’s disease (AD) is becoming more prevalent as the population ages, but there are no effective treatments for this devastating condition. Researchers developed a rhesus monkey model of AD by targeting the entorhinal cortex with an adeno-associated virus expressing mutant tau protein. Within 3 months they observed evidence of misfolded tau propagation, similar to what is hypothesized for AD patients. Treated monkeys developed robust alterations in AD core biomarkers in cerebrospinal fluid and blood. These results highlight the initial stages of tau seeding and propagation in rhesus macaques, a potentially powerful translational model with which to test new AD therapies.

Supported by ORIP (P51OD 011107) and NIA.

 

Evaluating a New Class of AKT/mTOR Activators for HIV Latency-Reversing Activity Ex Vivo and In Vivo

Gramatica, Andrea et al., Journal of Virology. 2021 Feb 3; Epub ahead of print.

https://doi.org/10.1128/JVI.02393-20

Activation of latent HIV-1 expression could benefit many HIV cure strategies. Researchers evaluated two AKT/mTOR activators, SB-216763 and tideglusib, as a potential new class of LRAs. The drugs reactivated latent HIV-1 present in blood samples from aviremic individuals on antiretroviral therapy without causing T cell activation or impaired effector function of cytotoxic T lymphocytes or NK cells. When tested in vivo in monkeys, tideglusib showed unfavorable pharmacodynamic properties and did not reverse SIV latency. The discordance between the ex vivo and in vivo results underscores the importance of developing novel LRAs that allow systemic drug delivery to relevant anatomical compartments.

Supported by ORIP (P51OD011092), NIAID, NIGMS, NIMH, NCI, amfAR Institute for HIV Cure Research, University of California San Francisco-Gladstone Institute of Virology & Immunology Center for AIDS Research, and James B. Pendleton Charitable Trust.

 

Virus Control in Vaccinated Rhesus Macaques Is Associated with Neutralizing and Capturing Antibodies against the SHIV Challenge Virus but Not with V1V2 Vaccine–Induced Anti-V2 Antibodies Alone

Hessell, Ann J. et al., Journal of Immunology. 2021 Mar 15; 206(6):1266-1283.

https://doi.org/10.4049/jimmunol.2001010

In the RV144 human immunodeficiency virus (HIV) vaccine trial, the only immune response associated with reduced infection was a high level of antibodies (Abs) targeting the second variable (V2) loop of the HIV envelope protein (Env). The mechanism underlying this suggested contribution of V2 Abs to protection remains unknown. Researchers tested the role of vaccine-induced anti-V2 Abs in rhesus macaques. Three vaccines strategies were designed to induce only V1V2 Abs before simian-human immunodeficiency virus (SHIV) challenge. Vaccine-induced V2 Abs did not independently control SHIV infection. However, neutralizing and virus capture anti-Env Abs were found to correlate with SHIV control.

Supported by ORIP (P51OD011092) and NIAID.

 

Immune Variations Throughout the Course of Tuberculosis Treatment and its Relationship with Adrenal Hormone Changes in HIV-1 Patients Co-Infected with Mycobacterium tuberculosis

Vecchione, María Belén et al., Tuberculosis. 2021 Mar; 127:102045.

https://doi.org/10.1016/j.tube.2020.102045

The probability of developing tuberculosis (TB) is 19 times higher in people infected with human immunodeficiency virus (HIV) compared to the general population. As host immune response defines the course of infection, researchers aimed to identify immuno-endocrine changes over six months of anti-TB chemotherapy in HIV+ people. Throughout the course of anti-TB/HIV treatment, plasma dehydroepiandrosterone (DHEA) and DHEA-sulfate levels increased while cortisol decreased. The balance between cortisol and DHEA, together with clinical assessment, served as a predictor of clinical outcome after anti-TB treatment. This research suggests that combined anti-HIV/TB therapies may partially restore both immune function and adrenal hormone levels.

Supported by ORIP (P51OD011133), Argentina’s National Agency for the Promotion of Research, Technological Development, and Argentine Innovation, and the University of Buenos Aires.

 

Natural Killer Cells Activated through NKG2D Mediate Lung Ischemia-Reperfusion Injury

Calabrese et al., J Clin Invest. 2021 Feb 1;131(3):e137047

Pulmonary ischemia-reperfusion injury (IRI) causes early mortality and has no effective therapies. While natural killer (NK) cells are innate lymphocytes capable of recognizing injured cells, their roles in acute lung injury are incompletely understood. Here, investigators demonstrated that NK cells were increased in frequency and cytotoxicity in 2 different IRI mouse models. They showed that NK cells trafficked to the lung tissue from peripheral reservoirs and were more mature within lung tissue. Acute lung ischemia-reperfusion injury was blunted in a NK cell–deficient mouse strain but restored with adoptive transfer of NK cells. In human lung tissue, NK cells were increased at sites of ischemia-reperfusion injury and activated NK cells were increased in prospectively-collected human bronchoalveolar lavage in subjects with severe IRI. These data support a causal role for recipient peripheral NK cells in pulmonary IRI via NK cell NKG2D receptor ligation. Therapies targeting NK cells may hold promise in acute lung injury. Supported by ORIP (1S10OD026940), NHLBI, NIDDK, and other sources.

 

Fructose Stimulated De Novo Lipogenesis Is Promoted by Inflammation

Jelena et al., Nat Metab. 2020 Oct; 2(10):1034-1045

Non-alcoholic fatty liver disease (NAFD) affects 30% of adult Americans. While NAFD starts as simple steatosis with little liver damage, its severe manifestation as non-alcoholic steatohepatitis (NASH) is a leading cause of liver failure, cirrhosis, and cancer. Fructose consumption is proposed to increase the risk of hepatosteatosis and NASH. Excessive intake of fructose causes barrier deterioration and low-grade endotoxemia. Using a mouse model, the study examined the mechanism of how fructose triggers these alterations and their roles in hepatosteatosis and NASH pathogenesis. The results demonstrated that microbiota-derived Toll-like receptor (TLR) agonists promote hepatosteatosis without affecting fructose-1-phosphate (F1P) and cytosolic acetyl-CoA. Activation of mucosal-regenerative gp130 signaling, administration of the YAP-induced matricellular protein CCN1 or expression of the antimicrobial peptide Reg3b (beta) counteract fructose-induced barrier deterioration, which depends on endoplasmic-reticulum stress and subsequent endotoxemia. Endotoxin engages TLR4 to trigger TNF production by liver macrophages, thereby inducing lipogenic enzymes that convert F1P and acetyl-CoA to fatty acid in both mouse and human hepatocytes. The finding may be of relevance to several common liver diseases and metabolic disorders. Supported by ORIP (S10OD020025), NCI, NIEHS, NIDDK, NIAID, and NIAAA.

 

Best Practices for Correctly Identifying Coronavirus by Transmission Electron Microscopy

Bullock et al., Kidney Int 2021 Apr;99(4):824-827

This paper provides strategies for identifying coronaviruses by transmission electron microscopy in ultrathin sections of tissues or tissue cultures. As illustrated by results in the literature, organ damage may be incorrectly attributed to the presence of virus, since images of coronavirus may resemble subcellular organelles. The paper also references numerous biochemical and imaging techniques to aid an investigator in avoiding pseudo positive identifications. ORIP grant support: S10OD026776

 

A chromosome-level genome of Astyanax mexicanus surface fish for comparing population-specific genetic differences contributing to trait evolution

Warren WC et al. Nat Commun. 2021 Mar 4;12(1):1447 (PMC7933363)

Identifying the genetic factors that underlie complex traits is central to understanding the mechanistic underpinnings of evolution. Cave-dwelling Astyanax mexicanus populations are well adapted to subterranean life and many populations appear to have evolved troglomorphic (morphological adaptation of an animal to living in the constant darkness of caves) traits independently, while the surface-dwelling populations can be used as a proxy for the ancestral form. Warren et al. present a high-resolution, chromosome-level surface fish genome, enabling the first genome-wide comparison between surface fish and cavefish populations. Using this resource, they performed quantitative trait locus (QTL) mapping analyses and found new candidate genes for eye loss (dusp26). They also generated the first genome-wide evaluation of deletion variability across cavefish populations to gain insight into this potential source of cave adaptation. The surface fish genome reference now provides a more complete resource for comparative, functional and genetic studies of drastic trait differences within a species.

Supported by ORIP (R24 OD011198), NIA, NICHD, NIGMS, NIDCR.

 

A Novel Tau-Based Rhesus Monkey Model of Alzheimer’s Pathogenesis

Beckman et al., Alzheimer’s & Dementia. 2021 Mar 18; Epub ahead of print

Alzheimer’s disease (AD) is becoming more prevalent as the population ages, but there are no effective treatments for this devastating condition. Promising findings in rodents have failed to translate into successful AD therapies. Researchers developed a new rhesus monkey model of AD by targeting the entorhinal cortex (ERC) with an adeno-associated virus expressing a double tau mutation. Within 3 months they observed evidence of misfolded tau propagation, similar to what is hypothesized for AD patients. Tau spreading was accompanied by a strong neuroinflammatory response driven by TREM2+ microglia. Treated monkeys also developed robust alterations in AD core biomarkers in cerebrospinal fluid and blood. These results highlight the initial stages of tau seeding and propagation in rhesus macaques, a potentially powerful translational model with which to develop and test new AD therapies. ORIP grant support: P51 OD011107.

 

A pulsatile release platform based on photo-induced imine-crosslinking hydrogel promotes scarless wound healing

Jian Zhang, et al., Nat Commun. 2021 Mar 15;12(1):1670 (https://pubmed.ncbi.nlm.nih.gov/33723267/)

Skin wound healing is a dynamic and interactive process involving the collaborative efforts of growth factors, extracellular matrix (ECM), and different tissue and cell lineages. Although accumulating studies with a range of different model systems have increased our understanding of the cellular and molecular basis underlying skin scar formation, they have not been effectively translated to therapy. Development of effective therapeutic approaches for skin scar management is urgently needed. In this study, team of investigators devise a water-oil-water double emulsion strategy to encapsulate proteins within a photo-crosslinkable poly-lactic-co-glycolic acid (PLGA) shell, which can produce microcapsules with pulsatile drug release kinetics after administration. The results show that pulsatile release of the TGF-β inhibitor can accelerate skin wound closure while suppressing scarring in murine skin wounds and large animal preclinical models, suggesting that it could be an effective approach to achieve scarless wound healing in skin. ORIP support grant: R01OD023700.

 

Resident Memory T Cells Form during Persistent Antigen Exposure Leading to Allograft Rejection

Abou-Daya et al.,   Sci Immunol., 2021 Mar 19;6(57):eabc8122..

It is not clear whether Tissue-resident memory T cells (TRM) function in organ transplants where cognate antigen persists. This is a key question in transplantation as T cells are detected long term in allografts. Investigators showed that antigen-specific and polyclonal effector T cells differentiated in the graft into TRM and subsequently caused allograft rejection. Graft TRM proliferated locally, produced interferon-γ upon restimulation, and their in vivo depletion attenuated rejection. The vast majority of antigen-specific and polyclonal TRM lacked phenotypic and transcriptional exhaustion markers. Single-cell analysis of graft T cells early and late after transplantation identified a transcriptional program associated with transition to the tissue-resident state that could serve as a platform for the discovery of therapeutic targets. Thus, recipient effector T cells differentiate into functional graft TRM that maintain rejection locally. Targeting these TRM could improve renal transplant outcomes. ORIP grant support: 1S10OD011925-01 and 1S10OD019942-01

 

Metabolomics analysis of follicular fluid coupled with oocyte aspiration reveals importance of glucocorticoids in primate periovulatory follicle competency

Ravisankar et al., 2021 https://www.nature.com/articles/s41598-021-85704-6

Assisted reproductive therapy in primates requires ovarian stimulation protocols, which result in multiple heterogeneous oocytes with variable capacity for fertilization, cleavage, and blastocyst formation. Recovered oocytes from rhesus macaque follicles (n=74 follicles) were fertilized in vitro and classified as failed to cleave, cleaved but arrested, or able to form blastocysts. Metabolomics analysis of the follicular fluid identified 60 metabolites that were different among embryo classifications; key was an increase in the intrafollicular ratio of cortisol to cortisone in the blastocyst group, which was associated with translocation of the glucocorticoid receptor, NR3C1. The data suggest a role for NR3C1 in the regulation of follicular processes, such as expansion of cumulus granulosa cells, via paracrine signaling. Supported by ORIP (P51OD011092) and NICHD.

 

The giant axolotl genome uncovers the evolution, scaling, and transcriptional control of complex gene loci

Schloissnig S. et al. Proc Natl Acad Sci U S A. 2021 Apr 13;118(15):e2017176118.

Vertebrates harbor recognizably orthologous gene complements but vary 100-fold in genome size. How chromosomal organization scales with genome expansion is unclear, and how acute changes in gene regulation, as during axolotl limb regeneration, occur in the context of a vast genome has remained a riddle. Here, Schloissnig et al. describe the chromosome-scale assembly of the giant, 32 Gb axolotl genome. Hi-C contact data revealed the scaling properties of interphase and mitotic chromosome organization. Analysis of the assembly yielded understanding of the evolution of large, syntenic multigene clusters, including the Major Histocompatibility Complex (MHC) and the functional regulatory landscape of the Fibroblast Growth Factor 8 (Axfgf8) region. The axolotl serves as a primary model for studying successful regeneration.

Supported by ORIP (R24 OD010435, P40 OD019794).

 

Cytomegaloviral Determinants of CD8⁺ T Cell Programming and RhCMV/SIV Vaccine Efficacy

Malouli et al., Science Immunology. 2021 Mar 25; 6(57):eabg5413

Cytomegalovirus (CMV)-based vaccine vectors were developed to leverage the ability of CMVs to elicit sustained CD4⁺ and CD8⁺ T cell responses with broad tissue distribution. The 68-1 rhesus cytomegalovirus (RhCMV) vectors that express simian immunodeficiency virus (SIV) inserts induce major histocompatibility complex E (MHC-E)- and MHC-II-restricted, SIV-specific CD8⁺ T cell responses. The contribution of this unconventional MHC restriction to RhCMV/SIV vaccine efficacy are poorly understood. Researchers demonstrated that these responses result from genetic rearrangements in 68-1 RhCMV that disrupt the function of eight immunomodulatory proteins encoded by the virus. Repair of each of these genes with either RhCMV or human CMV counterparts shifted responses to MHC-Ia-restricted, or MHC-Ia- and MHC-II-restricted, CD8 T cell responses, but repairing the RhCMV genes did not protect against SIV. These findings suggest that MHC-E-restricted CD8⁺ T cell responses may be critical to protection against SIV. ORIP grant support: U42 OD023038 and P51 OD011092.

 

Sensitive Tracking of Circulating Viral RNA Through All Stages of SARS-CoV-2 Infection

Huang et al., Journal of Clinical Investigation. 2021 Apr 1; 131(7):e146031

Circulating SARS-CoV-2 RNA could represent a more reliable indicator of infection than nasal RNA, but quantitative reverse transcription PCR (RT-qPCR) lacks diagnostic sensitivity for blood samples. Researchers developed a CRISPR-amplified, blood-based COVID-19 (CRISPR-ABC) assay to detect SARSCoV-2 in plasma. They evaluated the assay using samples from SARS-CoV-2-infected African green monkeys and rhesus macaques as well as from COVID-19 patients. CRISPR-ABC consistently detected viral RNA in the plasma of the experimentally infected primates from 1 to 28 days after infection. The increases in plasma SARS-CoV-2 RNA in the monkeys preceded rectal swab viral RNA increases. In the patient cohort, the new assay demonstrated 91.2% sensitivity and 99.2% specificity versus RT-qPCR nasopharyngeal testing, and it also detected COVID-19 cases with transient or negative nasal swab RT-qPCR results. These findings suggest that detection of SARS-CoV-2 RNA in blood by CRISPR-augmented RT-PCR could improve COVID-19 diagnosis, facilitate the evaluation of SARS-CoV-2 infection clearance, and help predict the severity of infection. ORIP grant support: P51 OD011104.

 

Polyfunctional Tier 2–Neutralizing Antibodies Cloned Following HIV-1 Env Macaque Immunization Mirror Native Antibodies in a Human Donor

Spencer, David A. et al., Journal of Immunology. 2021 Mar; 206(5):999-1012.

https://doi.org/10.4049/jimmunol.2001082

HIV vaccine efforts are limited by viral strain diversity and the shielding of neutralization epitopes on the viral envelope, yet isolation of broadly neutralizing antibodies from infected individuals suggests the potential for eliciting protective antibodies through vaccination. Researchers cloned 58 monoclonal antibodies (mAbs) from a rhesus monkey immunized with envelope glycoprotein immunogens from an HIV-1 clade C–infected volunteer. Twenty mAbs exhibited some neutralizing activity. Cloned mAbs targeting the V3 region and CD4 binding site were capable of tier 2 (i.e., moderate) neutralization. This study demonstrates partial recapitulation of the human donor’s humoral immune response through nonhuman primate vaccination.

Supported by ORIP (P51OD011092) and NIAID

 

Autologous Transplant Therapy Alleviates Motor and Depressive Behaviors in Parkinsonian Monkeys

Tao, Yunlong et al., Nature Medicine, 1 March 2021.

https://www.nature.com/articles/s41591-021-01257-1

Generation of induced pluripotent stem cells (iPSCs) enables standardized of dopamine (DA) neurons for autologous transplantation therapy to improve motor functions in Parkinson disease (PD). Adult male rhesus PD monkeys receiving autologous, but not allogenic, transplantation exhibited recovery from motor and depressive signs of PD over a 2-year period without immunosuppressive therapy. Mathematical modeling showed correlations between surviving DA neurons with PET signal intensity and behavior recovery regardless of autologous or allogeneic transplant, suggesting a predictive power of PET and motor behaviors for surviving DA neuron number. The results demonstrate favorable efficacy of the autologous transplant approach to treat PD. Supported by ORIP (P51OD011106) NINDS, and NICHD

 

Trim-Away Mediated Knock Down Uncovers a New Function for Lbh During Gastrulation of Xenopus laevis

Weir, Emma et al. Dev Biol. 2021 Feb; 470:74-83 (PMC7855437)

https://pubmed.ncbi.nlm.nih.gov/33159936/

The protein Lbh was identified as necessary for cranial neural crest cell migration in Xenopus. To investigate its role in embryonic events, the authors employed the technique "Trim-Away" to degrade this maternally deposited protein. Trim-Away utilizes the E3 ubiquitin ligase trim21 to degrade proteins targeted with an antibody. Early knockdown of Lbh in Xenopus results in defects in gastrulation that present with a decrease in fibronectin matrix assembly, an increase in mesodermal cell migration and decrease in endodermal cell cohesion. The technique is also effective on a second abundant maternal Protein Kinase C And Casein Kinase Substrate In Neurons 2.

Supported by ORIP (R24 OD021485), NIDCR

 

Immunogenic BNT162b Vaccines Protect Rhesus Macaques from SARS-CoV-2

Vogel et al., Nature. 2021 Feb 1. Online ahead of print

The preclinical development of two BNT162b vaccine candidates, which contain lipid-nanoparticle formulated nucleoside-modified mRNA encoding SARS-CoV-2 spike glycoprotein-derived immunogens, was performed in rhesus macaques at the Southwest National Primate Research Center (SNPRC). BNT162b1 encodes a soluble, secreted, trimerised receptor-binding domain. BNT162b2 encodes the full-length transmembrane spike glycoprotein, locked in its prefusion conformation. Prime/boost vaccination of rhesus macaques with BNT162b candidates elicits SARS-CoV-2 neutralizing antibody titers that are 8.2 to 18.2 times that of a SARS-CoV-2 convalescent human serum panel. The vaccine candidates protected macaques from SARS-CoV-2 challenge, with BNT162b2 protecting the lower respiratory tract from the presence of viral RNA and with no evidence of disease enhancement. The BNT162b2 vaccine recently received emergency use authorization from FDA and is being administered within the United States. The SNPRC is supported by ORIP P51OD011103.

 

Modified Vaccinia Ankara Vector-Based Vaccine Protects Macaques from SARS-CoV-2 Infection, Immune Pathology and Dysfunction in the Lung

Routhu, Nanda Kishore et al., Immunity. 2021 Feb; Epub ahead of print.

https://doi.org/10.1016/j.immuni.2021.02.001

Any SARS-CoV-2 vaccine may have limitations such as need for ultracold storage, poor induction of CD8⁺ T cell response, or lack of cross-reactivity with emerging strains. Thus, multiple vaccines may be needed to bring COVID-19 under control. Using rhesus macaques, researchers showed that a modified vaccinia Ankara (MVA) vector-based SARS-CoV-2 vaccine expressing prefusion-stabilized spike protein induced strong neutralizing antibody and CD8⁺ T cell responses. The vaccine protected macaques from SARS-CoV-2 infection as well as infection-induced inflammation and B cell abnormalities in the lung. These results are promising considering the excellent safety and performance of MVA vector-based vaccines for other pathogens.

Supported by ORIP (P51OD011132, S10OD026799) and NIAID

 

Lung Expression of Human Angiotensin-Converting Enzyme 2 Sensitizes the Mouse to SARS-CoV-2 Infection

Han, Kun et al., American Journal of Respiratory Cell and Molecular Biology. 2021 Jan; 64(1):79-88.

https://doi.org/10.1165/rcmb.2020-0354OC

A rapidly deployable mouse model that recapitulates a disease caused by a novel pathogen would be a valuable research tool during a pandemic. Researchers were able to produce C57BL/6J mice with lung expression of human angiotensin-converting enzyme 2 (hACE2), the receptor for SARS-CoV-2. They did so by oropharyngeal delivery of a recombinant human adenovirus type 5 expressing hACE2. The transduced mice were then infected with SARS-CoV-2. Thereafter, the mice developed interstitial pneumonia with perivascular inflammation, exhibited higher viral load in lungs compared to controls, and displayed a gene expression phenotype resembling the clinical response in lungs of humans with COVID-19.

Supported by ORIP (P51OD011104, R21OD024931), NHLBI, and NIGMS

 

SARS-CoV-2 Induces Robust Germinal Center CD4 T Follicular Helper Cell Responses in Rhesus Macaques

Lakshmanappa, Yashavanth Shaan et al., Nature Communications. 22 January 2021, Article 541. https://www.nature.com/articles/s41467-020-20642-x

SARS-CoV-2 infection in both sexes of rhesus macaques, either infused with convalescent plasma, normal plasma, or receiving no infusion, resulted in transient accumulation of pro-inflammatory monocytes and proliferating CD4 T follicular helper (T_fh) cells, which are critical for persistent antibody responses. CD4 helper cell responses skewed predominantly toward a T_h1 response in blood, lung, and lymph nodes. This skewing is important to note, as weak interferon responses observed in COVID patients could hamper effective antiviral antibody and CD8 T-cell responses. Collectively, the data show induction of GC responses in a rhesus model of mild COVID-19.

Supported by ORIP (P51OD011107 and P40OD010976) and NIAID

 

Myelin‐specific T Cells in Animals with Japanese Macaque Encephalomyelitis

Govindan, Aparna N. et al., Wiley Online Library, first published 13 January 2021, Vol. 8, Issue 2.

https://onlinelibrary.wiley.com/doi/10.1002/acn3.51303

Investigators characterized the CD4+ and CD8+ T cells in demyelinating Japanese macaque encephalomyelitis (JME) lesions in age‐ and sex‐matched macaques and discovered differences in expression of myelin antigen sequences in the T cell. Mapping myelin epitopes revealed a heterogeneity in T cell responses among JME animals, which are associated with a proinflammatory pathogenic role in multiple sclerosis (MS). These findings draw further parallels between JME and MS and support the hypothesis that JME and possibly MS are triggered by mechanisms involving myelin damage and not myelin epitope mimicry.

Supported by ORIP (P51OD011092) and NINDS

 

Antibody-Mediated Depletion of Viral Reservoirs is Limited in SIV-Infected Macaques Treated Early With Antiretroviral Therapy

Swanstrom, Adrienne E. et al., Journal of Clinical Investigation. 2021 Jan; Epub ahead of print.

https://doi.org/10.1172/JCI142421

Virus-specific strategies to target the latent HIV reservoir in individuals on combination antiretroviral therapy (cART) have been limited by inefficient induction of viral protein expression. Researchers used rhesus macaques to investigate an antibody-mediated reservoir targeting strategy, targeting the CD4 molecule rather than a viral protein, to deplete potential viral target cells irrespective of infection status. Despite profound CD4⁺ T cell depletion in blood and lymph nodes, time to viral rebound following cART cessation was not delayed in anti-CD4 treated animals compared with controls, likely due to the limited antibody-mediated cell depletion that occurred in rectal tissue and lymphoid follicles.

Supported by ORIP (R24OD010976), NCI, and NIAID

 

A Modular Master Regulator Landscape Controls Cancer Transcriptional Identity

Paul et al., Cell. 2021 Jan 21;184(2):334-351.e20

The mechanisms linking genomic alterations to transcriptional identity of cancer cells remain elusive. Integrative genomic analysis, using a network-based approach, identified 407 master regulator (MR) proteins responsible for canalizing the genetics of individual samples from 20 cohorts in The Cancer Genome Atlas into 112 transcriptionally distinct tumor subtypes. MR proteins could be further organized into 24 pan-cancer, MR block modules (MRBs), each regulating key cancer hallmarks and predictive of patient outcome in multiple cohorts. Of all somatic alterations detected in each individual sample, >50% were predicted to induce aberrant MR activity, yielding insight into mechanisms linking tumor genetics and transcriptional identity and establishing non-oncogene dependencies. Genetic and pharmacological validation assays confirmed the predicted effect of upstream mutations and MR activity on downstream cellular identity and phenotype. Thus, co-analysis of mutational and gene expression profiles identified elusive subtypes and provided testable hypothesis for mechanisms mediating the effect of genetic alterations. Supported by ORIP (S10OD012351 and S10OD021764), NCI, and other sources.

 

Increased Proviral DNA in Circulating Cells Correlates with Plasma Viral Rebound in SIV-Infected Rhesus Macaques after Antiretroviral Therapy Interruption

Ziani et al., J Virol. 2021 Jan 6;JVI.02064-20

Investigators longitudinally tracked dynamic decay of cell-associated viral RNA/DNA in systemic and lymphoid tissues in SIV-infected rhesus macaques on prolonged combined antiretroviral therapy (cART) to evaluate predictors of viral rebound after treatment cessation. Suppressive cART substantially reduced plasma SIV RNA, cell-associated unspliced, and multiply spliced SIV RNA to undetectable levels, yet viral DNA remained detectable in systemic tissues and lymphoid compartments throughout cART. A rapid increase of integrated proviral DNA in peripheral mononuclear cells was detected once cART was withdrawn, accompanied by the emergence of detectable plasma viral load. The increase of peripheral proviral DNA post cART interruption correlated with the emergence and degree of viral rebound. These results suggest that measuring total viral DNA in SIV infection may be a relatively simple surrogate marker of reservoir size, and may predict viral rebound after treatment interruption, and inform treatment strategies. Supported by ORIP (P51OD011104), NIAID and NICHD.

 

Thresholds for Post-Rebound SHIV Control after CCR5 Gene-Edited Autologous Hematopoietic Cell Transplantation

Cardozo-Ojeda et al., Elife. 2021 Jan 12;10:e57646

Investigators developed a mathematical model to project the minimum threshold of C-C chemokine receptor type 5 (CCR5) gene-edited cells necessary for a functional cure from HIV. This was based on blood T cell reconstitution and plasma simian-HIV (SHIV) dynamics from SHIV-1157ipd3N4-infected juvenile pig-tailed macaques that underwent autologous transplantation with CCR5 gene editing. The model predicts that viral control can be obtained following analytical treatment interruption (ATI) when: (1) transplanted hematopoietic stem and progenitor cells (HSPCs) are at least fivefold higher than residual endogenous HSPCs after total body irradiation and (2) the fraction of protected HSPCs in the transplant achieves a threshold (76–94%) sufficient to overcome transplantation-dependent loss of SHIV immunity. Under these conditions, if ATI is withheld until transplanted gene-modified cells engraft and reconstitute to a steady state, spontaneous viral control is projected to occur. Supported by ORIP (P51OD010425), NCATS and NIAID.

 

Severely Ill COVID-19 Patients Display Impaired Exhaustion Features in SARS-CoV-2-Reactive CD8+ T Cells

Kusnadi1 et al., Sci Immunol. 2021 Jan 21;6(55):eabe4782

How CD8+ T cells respond to SARS-CoV-2 infection is not fully known. Investigators reported on the single-cell transcriptomes of >80,000 virus-reactive CD8+ T cells, obtained using a modified Antigen-Reactive T cell Enrichment assay, from 39 COVID-19 patients and 10 healthy subjects. COVID-19 patient cells were segregated into two groups based on whether the dominant CD8+ T cell response to SARS-CoV-2 was “exhausted” or not. SARS-CoV-2-reactive cells in the exhausted subset were increased in frequency and displayed less cytotoxicity and inflammatory features in COVID-19 patients with mild compared to severe illness. In contrast, SARS-CoV-2-reactive cells in the dominant non-exhausted subset from patients with severe disease showed enrichment of transcripts linked to co-stimulation, pro-survival Nuclear Factor κB signaling, and anti-apoptotic pathways, suggesting the generation of robust CD8+ T cell memory responses in patients with severe COVID-19 illness. Overall, this single-cell analysis revealed substantial diversity in the nature of CD8+ T cells responding to SARS-CoV-2. Supported by ORIP (S10RR027366 and S10OD025052), NIAID, NHLBI, NIGMS, and other sources.

 

Endogenous Zebrafish Neural Cre Drivers Generated by CRISPR/Cas9 Short Homology Directed Targeted Integration

Almeida, M.P. et al., Scientific Reports 2021 January 18;11(1):1732 (PMC7813866)

https://www.nature.com/articles/s41598-021-81239-y

Almeida et al. previously reported precision targeted integration of reporter DNA in zebrafish using CRISPR/Cas9. Here, they isolated zebrafish Cre recombinase drivers. A 2A-Cre recombinase transgene with 48 bp homology arms was targeted into proneural genes ascl1b, olig2 and neurod1. They observed high rates of germline transmission from 10 to 100% (10% olig2; 20% neurod1; 100% ascl1b). The lines Tg(ascl1b-2A-Cre)^is75, Tg(olig2-2A-Cre)^is76, and Tg(neurod1-2A-Cre)^is77 expressed functional Cre recombinase in the cell populations. Results demonstrate Cre recombinase expression is driven by the native promoter and regulatory elements of targeted genes. This approach is a cost-effective method to generate cell type specific zebrafish Cre and CreERT2 drivers.

Supported by ORIP (R24 OD020166).

 

Deploying MMEJ using MENdel in Precision Gene Editing Applications for Gene Therapy and Functional Genomics

Martínez-Gálvez G. et al. Nucleic Acids Research. 2021 January 11;49(1):67-78 (PMC7797032)

https://academic.oup.com/nar/article/49/1/67/6030233

Gene-editing experiments commonly elicit the error-prone non-homologous end joining for DNA double-strand break (DSB) repair. Martinez-Galvez et al. compared three DSB repair prediction algorithms - MENTHU, inDelphi, and Lindel. MENTHU correctly identified 46% of all PreMAs available, a ∼2- and ∼60-fold sensitivity increase compared to inDelphi and Lindel, respectively.  The investigators report the new algorithm MENdel, a combination of MENTHU and Lindel, that achieves the most predictive coverage of homogeneous out-of-frame mutations. They suggest that the use of MENdel helps researchers use MMEJ at scale for reverse genetics screenings to be viable for nearly all loss-of-function based gene editing therapeutic applications.

Supported by ORIP (R24 OD020166), NIGMS.