Grantee Publications



The High Affinity Dopamine D2 Receptor Agonist MCL-536: A New Tool for Studying Dopaminergic Contribution to Neurological Disorders

Subburaju, Sivan et al. ACS Chemical Neuroscience. 2021 April 21;12(8):1428-1437 (PMC8426090)

The dopamine D2 receptor exists in two different states, D2high and D2low; the former is the functional form of the D2 receptor and associates with intracellular G-proteins. The D2 agonist [3H]MCL-536 has high affinity for the D2 receptor (Kd 0.8 nM) and potently displaces the binding of (R-(-)-N-n-propylnorapomorphine (NPA; Ki 0.16 nM) and raclopride (Ki 0.9 nM) in competition binding assays. The authors characterized [3H]MCL-536. [3H]MCL-536 as metabolically stable. In vitro autoradiography on transaxial and coronal brain sections showed specific binding of [3H]MCL-536.  [3H]MCL-536's unique properties make it a valuable tool for research on neurological disorders like Parkinson's disease or schizophrenia.
Supported by ORIP (R43 OD020186, R44 OD024615), NIMH

Genetic Basis For an Evolutionary Shift From Ancestral Preaxial to Postaxial Limb Polarity in Non-urodele Vertebrates

Trofka, Anna et al. Current Biology. 2021 September 28;S0960-9822(21)01250-1

In most tetrapod vertebrates, limb skeletal progenitors condense with postaxial dominance. Posterior elements (ulna and fibula) appear prior to their anterior counterparts (radius and tibia), followed by digit-appearance order with continuing postaxial polarity.  Recent fossil evidence suggests that preaxial polarity represents an ancestral rather than derived state. These authors report that 5'Hoxd (Hoxd11-d13) gene deletion in mouse is atavistic and uncovers an underlying preaxial polarity in mammalian limb formation. Evolutionary changes in Gli3R activity level - key in the fin-to-limb transition - appear to be fundamental to the shift from preaxial to postaxial polarity in formation of the tetrapod limb skeleton.
Supported by ORIP (P40 OD01979), NCI, OHSU Medical Research Foundation and Shriners Hospitals for Children.

Prior Infection With SARS-CoV-2 WA1/2020 Partially Protects Rhesus Macaques Against Re-infection With B.1.1.7 and B.1.351 Variants

Chandrashekar, Abishek et al., Science Translational Medicine. 2021 September 21;eabj2641 Online ahead of print 

Using the rhesus macaque model, researchers addressed whether natural immunity induced by the original SARS-CoV-2 WA1/2020 strain protects against re-challenge with B.1.1.7 and B.1.351, known as the alpha and beta variants of concern, respectively. The investigators infected rhesus macaques with WA1/2020 and re-challenged them on day 35 with WA1/2020 or with the alpha or beta variants. Natural immunity to WA1/2020 led to robust protection against re-challenge with WA1/2020, partial protection against beta, and an intermediate degree of protection against alpha. These findings have important implications for vaccination and public health strategies in the context of emerging SARS-CoV-2 variants of concern.
Supported by ORIP (P51OD011106), NCI, Bill & Melinda Gates Foundation, Janssen Vaccines & Prevention BV, Ragon Institute, Musk Foundation, and Massachusetts Consortium on Pathogen Readiness.


Persistence of Viral RNA in Lymph Nodes in ART-suppressed SIV/SHIV-infected Rhesus Macaques

Cadena, Anthony M. et al., Nature Communications. 2021 March 5;12(1):1474 

The long-lived viral reservoir is a key obstacle to curing HIV/AIDS, yet the features of that reservoir during antiretroviral therapy (ART) remain poorly understood. Researchers undertook a comprehensive analysis of the SIV/SHIV reservoir in multiple lymphoid and non-lymphoid tissues from SIV/SHIV-infected rhesus macaques suppressed with ART for one year. Their findings support a model in which the tissue viral reservoir is rapidly and broadly seeded early during acute infection. Viral RNA persists lymphoid tissues despite a long period of suppressive ART. Therefore, viral latency does not appear to be universally transcriptionally silent; the reservoir may include a spectrum of latency depths. 
Supported by ORIP (R01OD024917), NIAID, and Ragon Institute.


Tissue-specific Transcriptional Profiling of Plasmacytoid Dendritic Cells Reveals a Hyperactivated State in Chronic SIV Infection

Lee, Michelle Y.-H. et al., PLoS Pathogens. 2021 June 28;17(6):e1009674

Persistent immune activation is an obstacle to optimal health for people living with HIV. Using RNA-Seq, researchers investigated the immunostimulatory potential of plasmacytoid dendritic cells (pDCs) in chronic SIV infection in rhesus macaques. They observed that pDCs have highly activated profiles in these animals. In contrast, pDCs from SIV-infected sooty mangabeys (natural hosts for SIV) had expression profiles similar to uninfected animals. In chronically infected rhesus macaques, interferon alpha transcripts were readily detected in lymph node-homing pDCs, but not those from blood. Therefore, pDCs are a major producer of type-I interferon in chronic SIV infection and could be a useful immunotherapy target.
Supported by ORIP (R24OD010445, P51OD 011132, P51OD011092, S10OD026799) and NIAID.

Gut Germinal Center Regeneration and Enhanced Antiviral Immunity by Mesenchymal Stem/Stromal Cells

Weber, Marina G. et al., JCI Insight. 2021 June 22;6(12):e149033 

Researchers investigated the effects of mesenchymal stem/stromal cell (MSC) infusions on gut mucosal recovery, antiviral immunity, and viral suppression in SIV-infected rhesus macaques. MSC treatment heightened virus-specific responses and reduced viral load. Clearance of SIV-positive cells from gut mucosal effector sites was correlated with regeneration of germinal centers, restoration of follicular B cells and T follicular helper cells, and enhanced antigen presentation by viral trapping within the follicular dendritic cell network. These changes were associated with enhanced gene expression for type I/II interferon signaling, B cell proliferation, and interleukin 7. MSC treatment also activated metabolic pathways associated with enhanced immunity and viral reduction.
Supported by ORIP (P51OD011107), NIAID, and Brazil National Council for Scientific and Technological Development.


Comparative Cellular Analysis of Motor Cortex in Human, Marmoset and Mouse

Bakken, Trygve E., et al., 2021 October 6;(598):111-119

Investigators used high-throughput transcriptomic and epigenomic profiling of more than 450,000 single nuclei in humans, marmosets, and mice, to characterize the cellular makeup of the primary motor cortex (M1), which exhibits similarities that mirror evolutionary distance and are consistent between the transcriptome and epigenome. Despite the overall conservation, many species-dependent specializations are apparent. These results demonstrate the robust molecular foundations of cell-type diversity in M1 across mammals and point to the genes and regulatory pathways responsible for the functional identity of cell types and their species-specific adaptations. 
Supported by ORIP (P51OD010425), NIMH, NCATS, NINDS, and NIDA. 


Monoclonal Antibodies Protect Aged Rhesus Macaques from SARS-CoV-2-induced Immune Activation and Neuroinflammation

Verma, Anil et al., Cell Reports, 2021 November 5, Vol. 37, Issue 5 

In aged diabetic female rhesus macaques, prophylactic administration of neutralizing monoclonal antibodies (mAbs) effectively limits SARS-CoV-2 replication in both the upper and lower respiratory tract, and decreases immune activation, including reducing interferon-induced chemokines and limiting effector CD4 T cell influx into the cerebrospinal fluid. These protective mechanisms took place in the areas of the body targeted by the virus and may prevent adverse inflammatory consequences of SARS-CoV-2 infection in high-risk populations. 
Supported by ORIP (P51OD011107), NIAID and NIA. 


Improving Rigor and Reproducibility in Nonhuman Primate Research 

Bliss-Moreau, Eliza et al., American Journal of Primatology, 2021 September 20, Vol. 83, Issue 12  

Investigators across interdisciplinary fields consider approaches to enhance rigor and producibility in nonhuman primate (NHP) research. Given their similarity to humans, NHPs are often the animal model of choice for translational/preclinical biomedical research. However, availability of NHPs is limited. Hence, increased rigor is required to maximize the information gained from NHP studies. The co-authors consider approaches, such as, normative protocols, preregistration, data sharing, and how more extensive training in biostatics can enhance rigorous research in NHPs across biomedical disciplines. 
Supported by ORIP (P51OD011107, P51OD011106, P51OD011132, P51OD010425, P51OD011104, P51OD011092, and P51OD011133).


Comparative Cellular Analysis of Motor Cortex in Human, Marmoset, and Mouse

Bakken, Trygve, E., et al., Nature, 2021 October 6; 598, 111–119

Investigators used high-throughput transcriptomic and epigenomic profiling of more than 450,000 single nuclei in humans, marmosets, and mice, to characterize the cellular makeup of the primary motor cortex (M1), which exhibits similarities that mirror evolutionary distance and are consistent between the transcriptome and epigenome. Despite the overall conservation, many species-dependent specializations are apparent. These results demonstrate the robust molecular foundations of cell-type diversity in M1 across mammals and point to the genes and regulatory pathways responsible for the functional identity of cell types and their species-specific adaptations. 
Supported by ORIP (P51OD010425), NIMH, NCATS, NINDS, and NIDA. 


Collective Behavior Emerges from Genetically Controlled Simple Behavioral Motifs in Zebrafish

Harpaz, Roy, et al. Science Advances, 2021 October 8, Vol. 8, Issue 41;eabi7460.

Harpaz et al. report that zebrafish regulate their proximity and alignment with each other at early larval stages. Two visual responses (one measuring relative visual field occupancy and one accounting for global visual motion, account for emerging group behavior). Mutations in genes known to affect social behavior in humans perturb these reflexes in individual larval zebrafish and change their emergent collective behaviors.  Model simulations show that changes in these two responses in individual mutant animals predict well the distinctive collective patterns that emerge in a group. Hence, group behaviors reflect in part genetically defined primitive sensorimotor “motifs” evident in young larvae. 
Supported by ORIP (R43 OD024879, R44 OD024879), NINDS, NSF

TGF-β1 Signaling is Essential for Tissue Regeneration in the Xenopus Tadpole Tail

Nakamura, Makoto et al., Biochemical and Biophysical Research Communications, 2021 August 6, Vol. 565, Pages 91-96

Amphibians such as Xenopus tropicalis exhibit a remarkable capacity for tissue regeneration after traumatic injury. Nakamura et al. show that inhibition of TGF-β1 function prevents tail regeneration in Xenopus tropicalis tadpoles. CRISPR-mediated knock-out (KO) of tgfb1 retards tail regeneration; the phenotype of tgfb1 KO tadpoles can be rescued by injection of tgfb1 mRNA. Cell proliferation, critical for tissue regeneration, is downregulated in tgfb1 KO tadpoles;  tgfb1 KO reduces the expression of phosphorylated Smad2/3 (pSmad2/3). These results show that TGF-β1 regulates cell proliferation through the activation of Smad2/3. They propose that TGF-β1 plays a critical role in TGF-β receptor-dependent tadpole tail regeneration in Xenopus.
Supported by ORIP (P40 OD010997, R24 OD030008)


Deep Learning-based Framework for Cardiac Function Assessment in Embryonic Zebrafish from Heart Beating Videos 

Naderi, Amir Mohammad et al., Computers in Biology and Medicine, 2021 August, Vol. 135, p. 104565

Zebrafish is a powerful model system for a host of biological investigations, cardiovascular studies, and genetic screening. However, the current methods for quantifying and monitoring Zebrafish cardiac functions involve tedious manual work and inconsistent estimations. Naderi et al. developed a Zebrafish Automatic Cardiovascular Assessment Framework (ZACAF) based on a U-net deep learning model for automated assessment of cardiovascular indices, such as ejection fraction (EF) and fractional shortening (FS) from microscopic videos of wildtype and cardiomyopathy mutant zebrafish embryos. The framework could be widely applicable with any laboratory resources, and the automatic feature holds promise to enable efficient, consistent, and reliable processing and analysis capacity.
Supported by ORIP (R44 OD024874)


The Bowfin Genome Illuminates the Developmental Evolution of Ray-finned Fishes

Thompson, Andrew W., et al., Nature Genetics, 2021 September, Vol. 53, Issue 9, Pages 1373-1384.

The bowfin (Amia calva) is a ray-finned fish that possesses a unique suite of ancestral and derived phenotypes, which are key to understanding vertebrate evolution. The phylogenetic position of bowfin as a representative of neopterygian fishes, its archetypical body plan and its unduplicated and slowly evolving genome make bowfin a central species for the genomic exploration of ray-finned fishes. Here the authors present a chromosome-level genome assembly for bowfin that enables gene-order analyses, settling long-debated neopterygian phylogenetic relationships. These resources connect developmental evolution among bony fishes, further highlighting the bowfin's importance for illuminating vertebrate biology and diversity in the genomic era.
Supported by ORIP (R01 OD011116).


Limited Expansion of Human Hepatocytes in FAH/ RAG2-Deficient Swine

Nelson, Erek David, et al., Tissue Engineering – Part A, 2021 October 7, Online ahead of print.

The mammalian liver's regenerative ability has led researchers to engineer animals as incubators for expansion of human hepatocytes. Nelson et al. engineered immunodeficient swine to support expansion of human hepatocytes and identify barriers to their clinical application. Immunodeficient swine were engineered by knockout of the recombinase-activating gene 2 (RAG2) and fumarylacetoacetate hydrolase (FAH). Immature human hepatocytes (ihHCs) were injected into fetal swine by intrauterine cell transplantation (IUCT) at day 40 of gestation. They identified the mechanism of the eventual graft rejection by the intact NK cell population. They confirmed the presence of residual adaptive immunity in this model of immunodeficiency.
Supported by ORIP (U42 OD011140)


Negative Inotropic Mechanisms of β-cardiotoxin in Cardiomyocytes by Depression of Myofilament ATPase Activity without Activation of the Classical β-adrenergic Pathway

Lertwanakarn, Tuchakorn, et al., Scientific Reports, 2021 October 27, Issue 11, Page 21154

Beta-cardiotoxin (β-CTX) from the king cobra venom (Ophiophagus hannah) was previously proposed as a novel β-adrenergic blocker. However, the involvement of β-adrenergic signaling by this compound has never been elucidated. The objectives of this study were to investigate the underlying mechanisms of β-CTX as a β-blocker and its association with the β-adrenergic pathway. Healthy Sprague Dawley rats were used for cardiomyocytes isolation. In summary, the negative inotropic mechanism of β-CTX was discovered. β-CTX exhibits an atypical β-blocker mechanism. These properties of β-CTX may benefit in developing a novel agent aid to treat hypertrophic cardiomyopathy.
Supported by ORIP (P40 OD010960), NHLBI


Safety, Pharmacokinetics and Antiviral Activity of PGT121, a Broadly Neutralizing Monoclonal Antibody Against HIV-1: A Randomized, Placebo-Controlled, Phase 1 Clinical Trial

Stephenson, Kathryn et al., Nature Medicine. 2021 Oct;27(10):1718-1724. 

Researchers carried out a double-blind trial of one administration of the HIV-1 V3-glycan-specific antibody (Ab) PGT121 in HIV-uninfected and HIV-infected adults on antiretroviral therapy (ART), as well as an open-label trial of one infusion of PGT121 in viremic HIV-infected adults not on ART. The investigators observed no treatment-related serious adverse events among the 48 participants, and neutralizing anti-drug Abs were not elicited. PGT121 reduced plasma HIV RNA by a median of 1.77 log in viremic participants. Two individuals experienced ART-free viral suppression for ≥168 days following Ab infusion. These findings motivate further investigation of Ab-based therapeutic strategies for long-term HIV suppression. 
Supported by ORIP (R01OD024917, R01OD011095), NIAID, NCATS, and Bill and Melinda Gates Foundation.


Cytomegalovirus Mediates Expansion of IL-15-Responsive Innate-Memory Cells with SIV Killing Function

Méndez-Lagares, Gema et al., Journal of Clinical Investigation. 2021 Aug 2 ;131(15):e148542. 

Researchers investigated the effects of rhesus cytomegalovirus (RhCMV) on the immune system in young rhesus macaques to determine if it could modulate the protection mediated by RhCMV-vectored vaccines. RhCMV was associated with dramatic changes in antigen presenting cells, T cells, and NK cells and marked expansion of innate-memory CD8+ T cells via host interleukin-15 (IL-15) production. The researchers also investigated immune changes following administration of RhCMV 68-1–vectored SIV vaccines, which led to expansion of CD8+ T cells with capacity to inhibit SIV replication ex vivo. These results suggest that innate-memory expansion could be achieved by other vaccine platforms expressing IL-15.
Supported by ORIP (P51OD011107), NIAID, and California HIV/AIDS Research Foundation.


Immune Inactivation of Anti-Simian Immunodeficiency Virus Chimeric Antigen Receptor T Cells in Rhesus Macaques

Haeseleer, Françoise et al., Molecular Therapy–Methods & Clinical Development. 2021 Jun 24;22:304-319. 

Chimeric antigen receptor (CAR) T cell therapies are under development as potential HIV cures. Researchers found that CAR T cells expressing a single-chain variable fragment (scFv) that recognizes V1 or V3 of the SIV envelope eliminated SIV-infected T cells in vitro. However, in vivo infusion of these CAR T cells in rhesus macaques resulted in no detectable antiviral activity. Anti-SIV IgG antibodies in the SIV-infected animals were associated with inhibited CAR T cell effector functions. Thus, lack of in vivo efficacy of CAR T cells might be due to antibodies blocking the interaction between the CAR scFv and its epitope.
Supported by ORIP (P51OD011092), NIAID, Gilead Sciences HIV Cure Grant Program, and Juno Therapeutics.

Protection from SARS-CoV-2 Delta one year after mRNA-1273 vaccination in rhesus macaques coincides with anamnestic antibody response in the lung.

Gagne et al. in Cell.

Efficacy of the vaccine mRNA-1273 against SARS-CoV-2 Delta decreases with time, yet there are limited data on how durability of immune responses affects protection. Researchers immunized male rhesus macaques with mRNA-1273 and challenged them with Delta one year later. Serum neutralizing antibody responses to Delta and protection in upper airway were low one year after mRNA-1273 vaccination. However, mRNA-1273 provided durable protection against Delta in the lower airway and against severe lung disease one year after vaccination, likely through anamnestic induction of antibody responses in the lung. These findings highlight the importance of booster shots for sustained upper and lower airway protection. Supported by ORIP (P51OD011132), NIAID, Emory Executive Vice President for Health Affairs Synergy Fund, Woodruff Health Sciences Center, and Pediatric Research Alliance Center for Childhood Infections and Vaccines.


Cannabinoid Control of Gingival Immune Activation in Chronically SIV-Infected Rhesus Macaques Involves Modulation of the Indoleamine-2,3-Dioxygenase-1 Pathway and Salivary Microbiome

McDew-White, Marina et al., EBioMedicine. 23 December 2021;75:103769; online ahead of print.

HIV-associated periodontal disease (PD) affects people living with HIV (PLWH) on combination anti-retroviral therapy (cART). Researchers used a systems biology approach to investigate the molecular, metabolome, and microbiome changes underlying PD and its modulation by phytocannabinoids (Δ9-THC) in rhesus macaques. Δ9-THC reduced IDO1 protein expression. The findings suggest that phytocannabinoids may help reduce gingival/systemic inflammation, salivary dysbiosis, and potentially metabolic disease in PLWH on cART.


Dynamics and Origin of Rebound Viremia in SHIV-Infected Infant Macaques Following Interruption of Long-Term ART

Obregon-Perko, Veronica et al., JCI Insight. 8 December 2021;6(23):e152526.

Dynamics and Origin of Rebound Viremia in SHIV-Infected Infant Macaques Following Interruption of Long-Term ART

Researchers investigated viral rebound after analytical treatment interruption (ATI) in infant rhesus macaques infected with simian-human immunodeficiency virus (SHIV.C.CH505) and treated with long-term antiretroviral therapy (ART). Using a combination of ImmunoPET imaging, single-genome analysis of plasma SHIV RNA and cell-associated SHIV DNA, the researchers demonstrated that the gastrointestinal tract is a major site of viral RNA persistence that is capable of early SHIV.C.CH505 expansion following ATI, but virus reactivation in other tissues likely contributes to rebound viremia as ATI progresses.


CD4+ T Cells Are Dispensable for Induction of Broad Heterologous HIV Neutralizing Antibodies in Rhesus Macaques

Sarkar, Sanghita et al., Frontiers in Immunology. 20 October 2021;12:757811.

Researchers investigated the humoral response in vaccinated rhesus macaques with CD4+ T cell depletion, using the VC10014 DNA protein co-immunization vaccine platform (with gp160 plasmids and gp140 trimeric proteins derived from an HIV-1 infected subject).  Both CD4+-depleted and non-depleted animals developed comparable Tier 1 and 2 heterologous HIV-1 neutralizing plasma antibody titers. Thus, primates generate HIV neutralizing antibodies in the absence of robust CD4+ T cell help, which has important implications for vaccine development.

HDAC Inhibitor Titration of Transcription and Axolotl Tail Regeneration

Voss, S. Randal, et al., Frontiers in Cell and Development Biology. 31 December 2021; 31,9:76377. PMC8759488

New patterns of gene expression are enacted and regulated during tissue regeneration. Romidepsin, an FDA-approved HDAC inhibitor, potently blocks axolotl embryo tail regeneration by altering initial transcriptional responses to injury. Regeneration inhibitory concentrations of romidepsin increased and decreased the expression of key genes.  Single-nuclei RNA-Seq at 6 HPA illustrated  that key genes were altered by romidepsin in the same direction across multiple cell types. These results implicate HDAC activity as a transcriptional mechanism that operates across cell types to regulate the alternative expression of genes that associate with regenerative success versus failure outcomes.


Precise Visuomotor Transformations Underlying Collective Behavior in Larval Zebrafish

Harpaz, Roy et al., Nature Communications. 12 November 2021;12(1):6578. PMC8590009

Sensory signals from neighbors, analyzed in the visuomotor stream of animals, is poorly understood. The authors studied aggregation behavior in larval zebrafish and found that over development larvae transition from over dispersed groups to tight shoals. Young larvae turn away from virtual neighbors by integrating and averaging retina-wide visual occupancy within each eye, and by using a winner-take-all strategy for binocular integration. Observed algorithms accurately predict group structure over development. These findings allow testable predictions regarding the neuronal circuits underlying collective behavior in zebrafish.


Deep Learning is Widely Applicable to Phenotyping Embryonic Development and Disease

Naert, Thomas et al., Development. 05 November 2021;148(21):dev199664. PMC8602947

Genome editing simplifies the generation of new animal models for congenital disorders. The authors illustrate how deep learning (U-Net) automates segmentation tasks in various imaging modalities. They demonstrate this approach in embryos with polycystic kidneys (pkd1 and pkd2) and craniofacial dysmorphia (six1).  They provide a library of pre-trained networks and detailed instructions for applying deep learning to datasets and demonstrate the versatility, precision, and scalability of deep neural network phenotyping on embryonic disease models.

Genetic Control of the Pluripotency Epigenome Determines Differentiation Bias in Mouse Embryonic Stem Cells

Byers, Candice et al., The European Molecular Biology Organization. 2022 December 17;41(2): e109445. doi: 10.15252/embj.2021109445. Epub 2021 Dec 21. PMID: 34931323; PMCID: PMC8762565.

Investigators used derivation of mouse embryonic stem cells (ESC) to identify points or places regulating cell state transitions. This work demonstrated that ESCs derived from genetically diverse strains do not share equal developmental potential in vitro. Recent experiments have shown that differences in cell-fate choice during development may be critical in predisposing individuals to complex diseases due to underlying differences in cell-type composition. The BXD Resource at the Jackson Laboratory is supported by ORIP P40 OD011102 and awarded to Dr. Cathleen M. Lutz.


AAV Capsid Variants with Brain-wide Transgene Expression and Decreased Liver Targeting After Intravenous Delivery in Mouse and Marmoset

Goertsen, David et al., Nature Neuroscience. 09 December 2021. 25(1):106-115.

Genetic intervention is increasingly being explored as a therapeutic option for debilitating disorders of the central nervous system (CNS). This project focused on organ-specific targeting of adeno-associated virus (AAV) capsids after intravenous delivery. These results constitute an important step forward toward achieving the goal of engineered AAV vectors that can be used to broadly deliver gene therapies to the CNS in humans. Supported by ORIP U24 OD026638 and awarded to Dr. Guoping Feng at Massachusetts Institute of Technology, Cambridge, MA, USA.


AAV5 Delivery of CRISPR-Cas9 Supports Effective Genome Editing in Mouse Lung Airway

Liang, Shun-Qing et al., Molecular Therapy (Journal of the American Society of Gene Therapy). 2022 January 5;30(1):238-243. doi: 10.1016/j.ymthe.2021.10.023. Epub 2021 October 23. PMID: 34695545; PMCID: PMC8753568.

Genome editing in the lung has the potential to provide long-term expression of therapeutic protein to treat lung genetic diseases. Liang et al. illustrated that AAV5 can efficiently deliver CRISPR-Cas9 to mouse lung airways and was the first to achieve ∼20% editing efficiency in those airways. Results were confirmed through independent experiments at two different institutes. This highly efficient dual AAV platform will facilitate the study of genome editing in the lung and other tissue types. Supported by ORIP U42OD026645 and awarded to Drs. Jason D. Heaney, Mary E. Dickinson and William R. Lagor at Baylor College of Medicine, Houston, TX, USA.


Blastocyst Development after Fertilization with in Vitro Spermatids Derived from Nonhuman Primate Embryonic Stem Cells

Khampang, Sujittra et al., F & Science, ScienceDirect. Volume 2, Issue, 4, 2021 November, Pages 365-375. 

Rhesus macaque pluripotent stem cells were differentiated into spermatogenic germ cell linages and matured in vitro to form spermatids that were capable of fertilizing oocytes (female or germ cells involved in reproduction) by intracytoplasmic spermatid injection, i.e., the egg is fertilized outside the body and the sperm is injected through a needle into the egg. Successful in vitro preimplantation embryo development was observed in approximately 12% of zygotes. The data suggest potential mechanisms to address male infertility. Supported by ORIP (R21OD020182, R01OD028223 and P51OD011092).


Heritability of Social Behavioral Phenotypes and Preliminary Associations with Autism Spectrum Disorder Risk Genes in Rhesus Macaques: A Whole Exome Sequencing Study

Gunter, Chris et al., Autism Research (official journal of the International Society for Autism Research). 2022 January 29. doi: 10.1002/aur.2675. Epub ahead of print. PMID: 35092647.

Investigators quantified individual variation in social interactions among juvenile rhesus macaques (n= 212, both sexes) using both a standard macaque ethogram (a catalogue of animal behavior over time) and a macaque-relevant modification of the human Social Responsiveness Scale to study genetic influences on key aspects of social behavior and interactions. The analyses demonstrate that various aspects of juvenile social behavior exhibit significant genetic heritability, with quantitative genetic effects similar to the autism spectrum disorder (ASD) in human children. The significant genetic and sequencing data may be used to examine potential genetic associations with human ASD. Supported by ORIP (P51OD011132), NIGRI and NIMH. 


Reduced Infant Rhesus Macaque Growth Rates Due to Environmental Enteric Dysfunction and Association with Histopathology in the Large Intestine

Hendrickson, Sara M. et al., Nature Communications. 13, Article Number 234. Published 11 January 2022

Researchers characterized environmental enteric (relating to the intestines) dysfunction (EED) among infant rhesus macaques (n=80, both sexes) naturally exposed to enteric pathogens commonly linked to human growth stunting. Despite atrophy and abnormalities observed in the small intestine, poor growth trajectories and low serum tryptophan (an amino acid needed for protein and enzymes) levels were correlated with increased histopathology (microscopic tissue examination for disease manifestation) in the large intestine. This study provides insight into the mechanisms underlying EED and indicates that the large intestine may be an important target for therapeutic intervention. Supported by ORIP (P51OD011107 and P51OD011092) and the Bill and Melinda Gates Foundation. 

Functional and Ultrastructural Analysis of Reafferent Mechanosensation in Larval Zebrafish

Odstrcil, Iris, et al., Current Biology. 2022 January 10;32(1):176-189.e5. PMC8752774

All animals need to differentiate between exafferent stimuli (caused by the environment) and reafferent stimuli (caused by their own movement). Researchers characterized how hair cells in zebrafish larvae discriminate between reafferent and exafferent signals. Dye labeling of the lateral line nerve and functional imaging was combined with ultra-structural EM circuit reconstruction to show that cholinergic signals originating from the hindbrain transmit efference copies and dopaminergic signals from the hypothalamus may affect threshold modulation. Findings suggest that this circuit is the core implementation of mechanosensory reafferent suppression in these young animals. Supported by ORIP Small Business grants (R43OD024879, R44OD024879) and NINDS.


Natural Disaster and Immunological Aging in a Nonhuman Primate

Watowich, Marina M., et al., Proceedings of the National Academy of Sciences of the United States of America. 2022 February 22;119(8):e2121663119. PMC8872742

Weather-related disasters can exacerbate existing morbidities and increase mortality risk. Researchers examined Hurricane Maria’s impact on immune cell gene expression in large, age-matched, cross-sectional samples from free-ranging rhesus macaques (Macaca mulatta) living on an isolated island. Hurricane Maria was significantly associated with differential expression of 4% of immune-cell-expressed genes and were correlated with age-associated alterations in gene expression in addition to expression of key immune genes, dysregulated proteostasis networks, and greater expression of inflammatory immune cell-specific marker genes. These findings illuminate that natural disasters might become biologically embedded and contribute to earlier onset of disease and death.
Supported by ORIP (P40 OD012217), NIA, NIMH

Complex Decay Dynamics of HIV Virions, Intact and Defective Proviruses, and 2LTR Circles Following Initiation of Antiretroviral Therapy

White, Jennifer A. et al., Proc Natl Acad Sci USA. 2022 Feb 8;119(6):e2120326119. 

In people living with HIV-1 (PLWH) who start antiretroviral therapy (ART), virus in blood decreases rapidly to below detection, but remaining infected cells may become part of the latent reservoir. Researchers investigated viral decay dynamics and identified decay processes with pronounced differences between intact and defective proviruses. Infected cells that survive second-phase decay may down-regulate HIV-1 gene expression and enter the stable latent reservoir. This research provides insight into meaningful latent reservoir markers and mechanisms for elimination of cells with intact viral genomes.
Supported by ORIP (R01OD011095), NIAID, U.S. Department of Energy, Bill and Melinda Gates Foundation, and Howard Hughes Medical Institute.


Phagocytosis by an HIV Antibody Is Associated with Reduced Viremia Irrespective of Enhanced Complement Lysis

Spencer, David A. et al., Nature Communications. 2022 Feb 3;13(1):662. 

Researchers used the bNAb 10E8v4 targeting the HIV Env protein to examine the role of antibody-mediated effector and complement (C’) activity when 10E8v4 was administered prophylactically to rhesus monkeys challenged with simian-human immunodeficiency virus (SHIV). With sub-protective dosing, the researchers found a 78–88% reduction in post-acute viremia that was associated with 10E8v4-mediated phagocytosis. These results suggest that effector functions inherent to unmodified 10E8v4 contribute to therapeutic efficacy against SHIV, while C’ functions do not contribute to efficacy in this context. This research informs the design of bNAb modifications for improving the protective efficacy of this therapeutic approach against HIV.
Supported by ORIP (P51OD011092, U42OD023038) and NIAID.

Selective G protein signaling driven by substance P–neurokinin receptor dynamics.

Harris et al, in Nature Chemical Biology.


Investigators determined the cryogenic-electron microscopy structures of active neurokinin-1 receptor (NK1R) bound to neuropeptide substance P (SP) or the G protein q (Gq)-biased peptide SP6–11. Peptide interactions deep within NK1R are critical for receptor activation. Conversely, interactions between SP and NK1R extracellular loops are required for potent Gs-signaling but not Gq-signaling. Molecular dynamics simulations showed that these superficial contacts restrict SP flexibility. SP6–11, which lacks these interactions, is dynamic while bound to NK1R. Structural dynamics of NK1R agonists therefore depend on interactions with the receptor extracellular loops and regulate G protein signaling selectivity. This data unveils the molecular mechanism of how two stimuli (SP and Neurokinin A) yield distinct G protein signaling at the same G protein-coupled receptor. ORIP grant support: S10OD021741 (Talos Arctica 200kV cryogenic transmission electron microscope) and S10OD020054 (high performance Linux cluster for near atomic resolution single particle cryo-EM).


The Pigtail Macaque (Macaca nemestrina) Model of COVID-19 Reproduces Diverse Clinical Outcomes and Reveals New and Complex Signatures of Disease

Melton, Alexandra et al., PLoS Pathogens. 20 December 2021;17(12):e1010162.

Animal models that recapitulate human COVID-19 disease are critical for understanding SARS-CoV-2 viral and immune dynamics, mechanisms of disease, and testing of vaccines and therapeutics. A group of male pigtail macaques (PTM) were euthanized either 6- or 21-days after SARS-CoV-2 viral challenge and demonstrated mild-to-moderate COVID-19 disease. Pulmonary infiltrates were dominated by T cells, virus-targeting T cells were predominantly CD4+, increases in circulating inflammatory and coagulation markers, pulmonary pathologic lesions, and the development of neutralizing antibodies were observed. Collectively, the data suggests PTMs are a valuable model to study COVID-19 pathogenesis and may be useful for testing vaccines and therapeutics. Supported by ORIP (P51OD011104) and NIAID.


Antiretroviral Therapy Timing Impacts Latent Tuberculosis Infection Reactivation in a Tuberculosis/Simian Immunodeficiency Virus Coinfection Model

Sharan, Riti et al., Journal of Clinical Investigation. 2 December 2021;e153090.

In the rhesus macaque model of Mycobacterium tuberculosis + simian immunodeficiency virus (SIV) co-infection, chronic immune activation rather than depletion of CD4+ T cells correlates with reactivation of latent tuberculosis infection (LTBI). Researchers administered combined antiretroviral therapy (cART) at 2 weeks post-SIV co-infection to study if restoration of CD4+ T cell immunity occurred more broadly, and if this prevented LTBI compared to cART initiated at 4 weeks post-SIV. Earlier initiation of cART enhanced survival led to better control of viral replication and reduced immune activation in the periphery and lung vasculature, thereby reducing the rate of SIV-induced reactivation. Supported by ORIP (K01OD031898, P51OD011133, P51OD011132, S10OD028653) and NIAID.

Immune correlates analysis of the mRNA-1273 COVID-19 vaccine efficacy clinical trial.

Gilbert et al. in Science.

Investigators determined that antibodies are the correlate of protection in vaccinated individuals enrolled in the Moderna coronavirus efficacy phase 3 clinical trial. Vaccine recipients were assessed for neutralizing and binding antibodies as correlates of risk for COVID-19 disease and as correlates of protection. All markers were inversely associated with COVID-19 risk and directly associated with vaccine efficacy. These results help define immune marker correlates of protection and may guide approval decisions for messenger RNA (mRNA) COVID-19 vaccines and other COVID-19 vaccines. Supported by ORIP (S10OD028685).

Sexual dimorphic impact of adult-onset somatopause on life span and age-induced osteoarthritis.

Poudel et. al. in Aging Cell.

Osteoarthritis (OA) is a major cause of disability worldwide. In humans, the age-associated decline in growth hormone (GH) levels was hypothesized to play a role in the etiology of OA. Investigators studied the impact of adult-onset isolated GH deficiency (AOiGHD) on the life span and skeletal integrity in aged mice. Reductions in GH during adulthood were associated with extended life span and reductions in body temperature in female mice only. However, end-of-life pathology revealed high levels of lymphomas in both sexes, independent of GH status. Skeletal characterization revealed increases in OA severity in AOiGHD mice. In conclusion, while their life span increased, AOiGHD female mice’s health span was compromised by high-grade lymphomas and the development of severe OA. In contrast, AOiGHD males, which did not show extended life span, showed an overall low grade of lymphomas but exhibited significantly decreased health span, evidenced by increased OA severity. Supported by ORIP (S10 OD010751), which contributed to purchasing the MicroComputed Tomography System used in the study.

A noncoding RNA modulator potentiates phenylalanine metabolism in mice.

Li, et al. in Science

The role of long noncoding RNAs (lncRNAs) in phenylketonuria (PKU), i.e., an inherited disorder causing build-up of an amino acid causing brain problems, is unknown. Investigators demonstrated that the mouse lncRNA Pair and human lncRNA HULC associate with phenylalanine hydroxylase (PAH). Pair-knockout mice exhibited phenotypes that faithfully models human PKU, such as excessive blood phenylalanine (Phe), growth retardation, and progressive neurological symptoms. HULC depletion led to reduced PAH enzymatic activities in human induced pluripotent stem cell-differentiated hepatocytes, i.e., that have the capacity to self-renew by dividing. To develop a strategy for restoring liver lncRNAs, these investigators designed lncRNA mimics that exhibit liver enrichment. Treatment with these mimics reduced excessive Phe in Pair -/- and PAH R408W/R408W mice and improved the Phe tolerance of these mice. Supported by ORIP (S10OD012304) which contributed to purchasing the Orbitrap Elite High-Resolution Mass Spectrometer for Proteomics and Metabolomics.

Integrated spatial multiomics reveals fibroblast fate during tissue repair.

Foster et. al. in Proc Natl Acad Sci U S A

The function of regenerative medicine in wound healing remains elusive, partially because of how fibroblasts program and respond to injury remains unclear. Investigators presented a multimodal -omics platform for the comprehensive study of cell populations in complex tissue, which allowed characterization of cells involved in wound healing across time and space. Through integrated analysis of single cell chromatin landscapes and gene expression states, coupled with spatial transcriptomic profiling, fibroblast epigenomes were imputed with temporospatial resolution. This allowed revelation of potential mechanisms controlling fibroblast fate during migration, proliferation, and differentiation following skin injury, and reexamination of the canonical phases of wound healing. Supported by ORIP S10OD018220 which supported upgrading a High Throughput DNA Sequencer and S10OD010580 which contributed to purchasing a Nikon A1Rsi resonant spectral confocal microscope.


An NR2F1-specific agonist suppresses metastasis by inducing cancer cell dormancy.

Khalil BD et al. in The Journal of Experimental Medicine.

Researchers described the discovery of a nuclear receptor NR2F1 antagonist that specifically activates dormancy programs in malignant cells. Agonist treatment resulted in a self-regulated increase in NR2F1 mRNA and protein and downstream transcription of a novel dormancy program. This program led to growth arrest in multiple human cell lines as well as patient-derived organoids. This effect was lost when NR2F1 was knocked out. In mice, agonist treatment resulted in inhibition of lung metastasis of head and neck squamous cell carcinomas, even after cessation of the treatment. This work provides proof of principle supporting the use of NR2F1 agonists to induce dormancy as a therapeutic strategy to prevent metastasis. Supported by ORIP (S10OD018522 and S10OD026880).


Antibody-peptide epitope conjugates for personalized cancer therapy.

Zhang et al. in Cancer Res.

Antibody-peptide epitope conjugates (APEC) are a new class of modified antibody-drug conjugates that redirect T cell viral immunity against tumor cells. Investigators developed an experimental pipeline to create patient-specific APECs and identified new preclinical therapies for ovarian carcinoma. Using functional assessment of viral peptide antigen responses to common viruses like cytomegalovirus in ovarian cancer patients, a library of 192 APECs with distinct protease cleavage sequences was created using the anti-epithelial cell adhesion molecule (EpCAM) antibody. The streamlined and systemic approach includes assessing APEC function in vivo using a new zebrafish xenograft platform that facilitates high-resolution single cell imaging to assess therapy responses and then validating top candidates using traditional mouse xenograft studies and primary patient samples. This study develops a high-throughput preclinical platform to identify patient-specific antibody-peptide epitope conjugates that target cancer cells and demonstrates the potential of this immunotherapy approach for treating ovarian carcinoma. Supported by ORIP (R24OD016761).


Complement Blockade in Recipients Prevents Delayed Graft Function and Delays Antibody-mediated Rejection in a Nonhuman Primate Model of Kidney Transplantation.

Eerhart et al. in Transplantation.

Investigators evaluated the efficacy of a high-dose recombinant human C1 esterase inhibitor (rhC1INH) in preventing delayed graft function (DGF) in a rhesus macaque (RM) model of kidney transplantation after brain death and prolonged cold ischemia. The majority (4 of 5) of vehicle-treated recipients developed DGF, whereas DGF was observed in only 1 of 8 rhC1INH-treated recipients. RM treated with rhC1INH also had faster creatine recovery, superior urinary output, and reduced biomarkers of allograft injury for the first week. The results suggest high-dose C1INH treatment in transplant recipients is an effective strategy to reduce kidney injury and inflammation, prevent DGF, delay antibody-mediated rejection development, and improve transplant outcomes. Supported by ORIP (P51OD011106) NIAID, and NIDDK.


Deciphering the role of mucosal immune responses and the cervicovaginal microbiome in resistance to HIV infection in HIV-exposed seronegative women

Ponnan et al., Microbiology Spectrum. 2021 Oct 31;9(2):e0047021.

Identifying correlates of protection in HIV-exposed seronegative (HESN) individuals requires identification of HIV-specific local immune responses. Researchers performed a comprehensive investigation of the vaginal mucosa and cervicovaginal microbiome in HESN women. They found elevated antiviral cytokines, soluble immunoglobulins, activated NK cells, CXCR5+ CD8+ T cells, and T follicular helper cells in HESN women compared to HIV-unexposed healthy women. They also found greater bacterial diversity and increased abundance of Gardnerella species in the mucosa of HESN women. These findings suggest that the genital tract of HESN women contains innate immune factors, antiviral mediators, and T cell subsets that protect against HIV. Supported by ORIP (P51OD011132), NIAID, Indian Department of Health Research, Indian Council of Medical Research, and Swedish Research Council.


A large repertoire of B cell lineages targeting one cluster of epitopes in a vaccinated rhesus macaque

Li et al., Vaccine. 2021 Sep 15;39(39):5607-5614.

A rhesus macaque that was serially immunized six times with the 8-mer epitope for human monoclonal antibody (mAb) 447-52D—specific to the V3 region of gp120 HIV-1—provided a rare opportunity to study the repertoire of antibodies produced upon vaccination against a particular antigenic site. From a blood sample taken 3 weeks after the last immunization, researchers produced 41 V3-specific recombinant mAbs by single B cell isolation and cloning. Sequence analysis revealed 21 B cell lineages (single and clonally related). The broad repertoire of Abs directed to a small antigenic site shows the targeting potency of a vaccine-elicited immune response in rhesus macaques. Supported by ORIP (P51OD011092, U42OD010246) and NIAID.


A novel wireless ECG system for prolonged monitoring of multiple zebrafish for heart disease and drug screening studies

Le T et al. Biosens Bioelectron. 2022 Feb 1;197:113808. doi: 10.1016/j.bios.2021.113808. Epub 2021 Nov 16.

Zebrafish and their mutant lines have been extensively used in cardiovascular studies. In the current study, the novel system, Zebra II, is presented for prolonged electrocardiogram (ECG) acquisition and analysis for multiple zebrafish within controllable working environments. The Zebra II is composed of a perfusion system, apparatuses, sensors, and an in-house electronic system. First, the Zebra II is validated in comparison with a benchmark system, namely iWORX, through various experiments. The validation displayed comparable results in terms of data quality and ECG changes in response to drug treatment. The effects of anesthetic drugs and temperature variation on zebrafish ECG were subsequently investigated in experiments that need real-time data assessment. The Zebra II's capability of continuous anesthetic administration enabled prolonged ECG acquisition up to 1 h compared to that of 5 min in existing systems. The novel, cloud-based, automated analysis with data obtained from four fish further provided a useful solution for combinatorial experiments and helped save significant time and effort. The system showed robust ECG acquisition and analytics for various applications including arrhythmia in sodium induced sinus arrest, temperature-induced heart rate variation, and drug-induced arrhythmia in Tg(SCN5A-D1275N) mutant and wildtype fish. The multiple channel acquisition also enabled the implementation of randomized controlled trials on zebrafish models. The developed ECG system holds promise and solves current drawbacks in order to greatly accelerate drug screening applications and other cardiovascular studies using zebrafish.
Supported by ORIP (R44 OD024874), NHLBI, NSF


Challenges and Considerations during In Vitro Production of Porcine Embryos

Chen PR et al. Cells. 2021 Oct 15;10(10):2770. PMC8535139.

Genetically modified pigs have become valuable tools for generating advances in animal agriculture and human medicine. Importantly, in vitro production and manipulation of embryos is an essential step in the process of creating porcine models. As the in vitro environment is still suboptimal, it is imperative to examine the porcine embryo culture system from several angles to identify methods for improvement. Understanding metabolic characteristics of porcine embryos and considering comparisons with other mammalian species is useful for optimizing culture media formulations. Furthermore, stressors arising from the environment and maternal or paternal factors must be taken into consideration to produce healthy embryos in vitro. In this review, Chen et al progress stepwise through in vitro oocyte maturation, fertilization, and embryo culture in pigs to assess the status of current culture systems and address points where improvements can be made.
Supported by ORIP (U42 OD011140), USDA


Publications Archive




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

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

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

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

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

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 SHIVSF162P3-Infected Macaques Treated with Anti-HIV Broadly Neutralizing Antibodies

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

To explore therapeutic potentials of combining anti-HIV broadly neutralizing antibodies (bNAbs) with α4β7 integrin blockade using the monoclonal antibody Rh-α4β7, investigators treated SHIVSF162P3-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 SHIVSF162P3-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.

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

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.

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

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

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

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

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.

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.

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.

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.

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).

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.

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.

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 (SHIVSF162P3). 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

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

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

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.

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.

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.

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.

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.

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.

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

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

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

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.

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.

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.

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

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

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

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

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

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

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

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

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

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)

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:

(Article’s Editorial commentary:

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

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

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

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

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.

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).

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.

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

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

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

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

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

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.

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

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.

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)

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

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.

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.

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.

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.

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 (

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

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.

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.

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)

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.

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.

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.

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 (Tfh) cells, which are critical for persistent antibody responses. CD4 helper cell responses skewed predominantly toward a Th1 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.

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.

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)

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)

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.



The Immune Landscape in Tuberculosis Reveals Populations Linked to Disease and Latency

Esaulova et al., Cell Host Microbe. 2020 Dec 16;S1931-3128(20)30635-1

Mycobacterium tuberculosis infection of adult rhesus macaques (RMs), predominantly males (81%), recapitulates both latent (LTBI) and active pulmonary TB (PTB) observed in humans. The immune characterization in lungs of RMs with PTB exhibited an influx of plasmacytoid dendritic cells, an interferon-responsive macrophage population, and activated T cell responses. In contrast, a CD27+ natural killer (NK) cell subset accumulated in the lungs of RMs with LTBI. This NK cell population was also detected in the circulation of humans with LTBI. This characterization of lung immune cells enhances our understanding of TB immunopathogenesis and provides potential targets for therapies and vaccines for TB control. Supported by ORIP (P51OD011104 and P51OD011133), NHLBI and NIAID.


A Frog with Three Sex Chromosomes that Co-Mingle Together in Nature: Xenopus tropicalis Has a Degenerate W and a Y that Evolved from a Z Chromosome

Furman, Benjamin L. S., et al. PLoS Genet. 2020 Nov 9; 16(11):e1009121 (PMC7652241)

Genetic systems governing sexual differentiation vary among species. Furman et al. investigated a frog with three sex chromosomes, the Western clawed frog, Xenopus tropicalis. They demonstrate that natural populations from the western and eastern edges of Ghana have a young Y chromosome, and that a male-determining factor on this Y chromosome is in a similar genomic location as a previously known female-determining factor on the W chromosome. Their findings are consistent with theoretical expectations associated with recombination suppression on sex chromosomes and demonstrate that several characteristics of old and established sex chromosomes can arise well before they become cytogenetically distinguished.

Supported by ORIP (P40 OD010997), NICHD


3-D Printed Customizable Vitrification Devices for Preservation of Genetic Resources of Aquatic Species

Tiersch, Connor J., et al. Aquac Eng. 2020 Aug; 90:102097 (PMC7434064)

Sperm vitrification as an alternative approach to conventional cryopreservation allows quick and low-cost sample preservation and is suitable for small-bodied aquatic species with miniscule testis, fieldwork at remote locations, and small-scale freezing for research purposes. Tiersch et al. report the developing of operational prototypes of 3-dimensional (3-D) printed vitrification devices.  This study demonstrated the feasibility of developing standardized low-cost devices fabricated by 3-D printing with functions including vitrification, volume control, labeling, protection, and storage. These prototypes can be further developed to assist development of germplasm repositories to protect the genetic resources of aquatic species by breeders, hatcheries, aquariums, and researchers.

Supported by ORIP (R24 OD010441)


Sequence Diversity Analyses of an Improved Rhesus Macaque Genome Enhance its Biomedical Utility

Warren et al., Science. 18 December 2020: Vol. 370, Issue 6523, eabc6617

Investigators sequenced and assembled an Indian-origin female rhesus macaque (RM) genome using a multiplatform genomics approach that included long-read sequencing, extensive manual curation, and experimental validation to generate a new comprehensive annotated reference genome. As a result, 99.7% of the gaps in the earlier draft genome are now closed, and more than 99% of the genes are represented. Whole-genome sequencing of 853 RMs of both sexes identified 85.7 million single-nucleotide variants and 10.5 million indel variants, including potentially damaging variants in genes associated with human autism and developmental delay. The improved assembly of segmental duplications, new lineage-specific genes and expanded gene families provide a framework for developing noninvasive NHP models of human disease as well as studies of genetic variation and phenotypic consequences. Supported by ORIP (P51OD011106, P51OD011107, P51OD011132, P51OD011104, U42OD024282, U42OD010568, R24OD011173, R24OD021324, and R24OD010962), NHGRI, NIMH, NHLBI, and NIGMS.


Biological Activities of a New Crotamine-like Peptide from Crotalus Oreganus Helleri on C2C12 and CHO Cell Lines, and Ultrastructural Changes on Motor Endplate and Striated Muscle

Salazar et al. Toxicon. 2020 December;188:95-107 (PMC7720416)

Crotamine and crotamine-like peptides are non-enzymatic polypeptides found in high concentration in the Crotalus genus venom. Helleramine was isolated and purified from the venom of the rattlesnake, Crotalus oreganus helleri. Purified helleramine increased intracellular Ca2+ in Chinese Hamster Ovary (CHO) cell line, inhibited cell viability of C2C12 (immortalized skeletal myoblast) and promoted early apoptosis and cell death. Skeletal muscle harvested from mice 24 h after helleramine injection showed contracted myofibrils and profound vacuolization, with loss of plasmatic and basal membrane integrity. The effects of helleramine provide evidence of myotoxic activities of crotamine-like peptides and their possible role in crotalid envenoming.

Supported by ORIP (P40 OD010960).

Responses to Acute Infection with SARS-CoV-2 in the Lungs of Rhesus Macaques, Baboons and Marmosets
Singh, Dhiraj K. et al., Nature Microbiology, 18 December 2020
Investigators compared acute SARS-CoV-2 infection in young and old rhesus macaques and baboons. Macaques had clinical signs of viral infection, mild to moderate pneumonitis and extra-pulmonary pathologies; both age groups recovered within two weeks. Baboons had prolonged viral RNA shedding and more lung inflammation compared with macaques; inflammation in bronchoalveolar lavage was increased in old versus young baboons. Macaques developed T-cell memory responses and bystander cytokine production. Old macaques had lower titers of SARS-CoV-2-specific IgG antibody levels compared with young macaques. The results indicate macaques and baboons experience acute respiratory distress that recapitulates the progression of COVID-19 in humans. 
Supported by ORIP (P51OD111033 and U42OD010442) and NIAID.


Imbalance of Regulatory and Cytotoxic SARS-CoV-2-Reactive CD4+ T Cells in COVID-19

Meckiff, Cell. 2020 Nov 25;183(5):1340-1353.e16.  

It is not clear why COVID-19 is deadly in some people and mild in others. To understand the underlying mechanism, investigators studied the contribution of CD4+ T cells in immune responses to SARS-CoV-2 infection. They analyzed single-cell transcriptomic data of >100,000 viral antigen-reactive CD4+ T cells from 40 COVID-19 patients. In hospitalized patients compared to non-hospitalized patients, they found increased proportions of cytotoxic follicular helper cells (TFH) and cytotoxic T helper (TH) cells responding to SARS-CoV-2 and reduced proportion of SARS-CoV-2-reactive regulatory T cells (TREG). Importantly, in hospitalized COVID-19 patients, a strong cytotoxic TFH response was observed early in the illness, which correlated negatively with antibody levels to SARS-CoV-2 spike protein. Polyfunctional TH1 and TH17 cell subsets were underrepresented in the repertoire of SARS-CoV-2-reactive CD4+ T cells compared to influenza-reactive CD4+ T cells. Together, these analyses provided insights into the gene expression patterns of SARS-CoV-2-reactive CD4+ T cells in distinct disease severities. Supported by ORIP  (S10RR027366 and S10OD025052), NIAID, NHLBI, NIGMS and other sources.


Infant Isoflurane Exposure Affects Social Behaviours, but Does Not Impair Specific Cognitive Domains in Juvenile Non-human Primates
Neudecker, Viola et al., British Journal of Anaesthesia 14 November 2020
Researchers investigated the impact of extended (5 hours) isoflurane anesthetic exposure (1-3 exposures) of rhesus macaque (RM) infants of both sexes on cognitive testing and behavioral assessments. Cognitive function did not differ among groups; however, compared to controls, RMs exposed three times during infancy exhibited less close social behavior. One isoflurane exposure resulted in increased anxiety-related behaviors and more inhibition towards novel objects. These findings are consistent with behavioral alterations observed in social settings of human clinical studies. 
Supported by ORIP (P51OD011092). 


Estrogen Acts Through Estrogen Receptor 2b to Regulate Hepatobiliary Fate During Vertebrate Development
Chaturantabut, Saireudee et al. Hepatology. 2020 Nov;72(5):1786-1799
During liver development, bipotent progenitor cells differentiate into hepatocytes and biliary epithelial cells to ensure a functional liver. The developmental cues controlling the differentiation of committed progenitors into these cell types are not completely understood. These authors report an essential role for estrogenic regulation in vertebrate liver development to affect hepatobiliary fate decisions. The studies identify17β-estradiol (E2), nuclear estrogen receptor 2b (esr2b), and downstream bone morphogenetic protein (BMP) activity as important regulators of hepatobiliary fate decisions during vertebrate liver development. These results have significant implications for liver development in infants exposed to abnormal estrogen levels or estrogenic compounds during pregnancy.
Supported by ORIP (R24 OD017870), NIDDK.


Lipocalin-2 Is an Anorexigenic Signal in Primates

Petropoulou, Peristera-Ioanna et al., eLife. 2020 Nov; 9:e58949.

The hormone Lipocalin-2 (LCN2) suppresses food intake in mice. Researchers demonstrated that LCN2 increases after a meal and reduces hunger in people with normal weight or overweight, but not in obese individuals. The researchers also showed that LCN2 crosses the blood-brain barrier and binds to the hypothalamus in vervet monkeys. LCN2 was found to bind to the hypothalamus in human, baboon, and rhesus macaque brain sections. When injected into vervets, LCN2 suppressed food intake and lowered body weight without toxic effects in short-term experiments. These findings lay the groundwork to investigate whether LCN2 might be a useful treatment for obesity.

Supported by ORIP (P40OD010965), NCATS, NIDDK, NIA, NHLBI, and National Institute of Food and Agriculture.


Antiretroviral Therapy Does Not Reduce Tuberculosis Reactivation in a Tuberculosis-HIV Coinfection Model

Ganatra, Shashank R. et al., Journal of Clinical Investigation. 2020 October;130(10):5171-5179

Despite treatment of HIV with antiretroviral therapy (ART), the risk of tuberculosis (TB) reactivation is higher in HIV-infected than HIV-uninfected persons. Researchers used Mycobacterium tuberculosis/SIV-coinfected rhesus macaques to model the impact of ART on TB reactivation due to HIV-induced immunosuppression. ART significantly reduced viral loads and increased CD4+ T-cell counts in blood, spleen, and bronchoalveolar lavage samples, but it did not reduce the risk of SIV-induced TB reactivation during the early phase of treatment. This study offers a translational model for the investigation of TB/SIV coinfection and the evaluation of treatment regimens to prevent TB reactivation in HIV-infected individuals.

Supported by ORIP (P51OD011133, P51OD011132) and NIAID.


Intra-Strain Genetic Variation of Platyfish (Xiphophorus maculatus) Strains Determines Tumorigenic Trajectory
Lu, Yuan et al., Frontiers in Genetics 2020 Oct 6;11:562594
Xiphophorus interspecies hybrids represent a valuable model system to study heritable tumorigenesis. Although the ancestors of the two X. maculatus parental lines, Jp163 A and Jp163 B, were siblings produced by the same mother, backcross interspecies hybrid progeny between X. hellerii and X. maculatus Jp163 A develop spontaneous melanoma initiating at the dorsal fin due to a regulator encoded by the X. maculatus genome; the backcross hybrid progeny with X. hellerii or X. couchianus and Jp163 B exhibit melanoma on their flanks. Comparative genomic analyses revealed genetic differences are associated with pathways highlighting fundamental cellular functions. Disruption of these baselines may give rise to spontaneous or inducible tumorigenesis.
Supported by ORIP (R24 OD-011120), NCI, NIGMS.


Induction and Characterization of Pancreatic Cancer in a Transgenic Pig Model
Boas, F. Edward et al., PLoS One. 2020 Sep 21;15(9):e0239391
Preclinical testing of new therapies for pancreatic cancer has been challenging due to lack of a suitable large animal model. Pigs, however, have similar physiology and immune response to humans. Boas et al report the development of a porcine model of pancreatic cancer.  H&E and immunohistochemical stains revealed undifferentiated carcinomas, like those of human pancreatobiliary systems. In several pigs, angiographies revealed that the artery supplying the pancreatic tumor could be catheterized using a 2.4 F microcatheter. In summary, pancreatic cancer can be induced in a transgenic pig, and intra-arterial procedures using catheters designed for human interventions were feasible in this model. 
Supported by ORIP (U42 OD011140), NCI.


Epidemiological and Molecular Characterization of a Novel Adenovirus of Squirrel Monkeys after Fatal Infection during Immunosuppression

Rogers, Donna L. et al. Microb Genom. 2020 Sep; 6(9):mgen000395 (PMC7643968)

Adenoviruses frequently cause upper respiratory tract infections often causing disseminated disease in immunosuppressed patients. A novel adenovirus was identified, squirrel monkey adenovirus 1 (SqMAdV-1), as the cause of a fatal infection in an immunocompromised squirrel monkey (Saimiri boliviensis). A nucleotide polymorphism at the stop codon of the DNA polymerase gene results in a 126 amino acid extension at the carboxy terminus. A single adenovirus variant, SqMAdV-3, has similarity to tufted capuchin (Sapajus apella) adenoviruses. The largest group of adenovirus variants detected, SqMAdV-2.0-2.16, has high similarity (93-99 %) to the TMAdV, suggesting that squirrel monkeys may be the natural host of the TMAdV.

Supported by ORIP (P40 OD010938, R24 OD018553), NIAID


Germline Transmission of Donor, Maternal and Paternal mtDNA in Primates
Ma, Hong et al, 2020; 
Mitochondrial gene mutations contribute to incurable human disorders. The possibility of using mitochondrial replacement therapy (MRT) to prevent transmission of pathogenic mitochondrial (mt)DNA was explored in rhesus macaques. Development of spindle MRT transfer in oocytes in 5 female rhesus macaques resulted in healthy and fertile offspring.  These results demonstrate that MRT is compatible with normal postnatal development including overall health and reproductive fitness in nonhuman primates with no detected adverse effects. Additional research is needed to more fully explore the use of MRT to prevent disorders as this study had a limited number of animals with only one female offspring. 
Supported by ORIP (P51OD0092) and NIA.

Last updated: 03-16-2022