Selected Grantee Publications
BNT162b Vaccines Protect Rhesus Macaques from SARS-CoV-2
Vogel et al., Nature. 2021.
https://www.nature.com/articles/s41586-021-03275-y
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).
Natural Killer Cells Activated Through NKG2D Mediate Lung Ischemia-Reperfusion Injury
Calabrese et al., Journal of Clinical Investigation. 2021.
https://www.jci.org/articles/view/137047
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 (S10OD026940), NHLBI, and NIDDK.
Trim-Away Mediated Knock Down Uncovers a New Function for Lbh During Gastrulation of Xenopus laevis
Weir et al., Developmental Biology. 2021.
https://pubmed.ncbi.nlm.nih.gov/33159936/
The protein Lbh was identified as necessary for cranial neural crest cell migration in Xenopus. To investigate its role in embryonic events, the authors employed the technique "Trim-Away" to degrade this maternally deposited protein. Trim-Away utilizes the E3 ubiquitin ligase trim21 to degrade proteins targeted with an antibody. Early knockdown of Lbh in Xenopus results in defects in gastrulation that present with a decrease in fibronectin matrix assembly, an increase in mesodermal cell migration and decrease in endodermal cell cohesion. The technique is also effective on a second abundant maternal Protein Kinase C And Casein Kinase Substrate In Neurons 2. Supported by ORIP (R24OD021485) and NIDCR.
Germline Transmission of Donor, Maternal and Paternal mtDNA in Primates
Ma et al., Human Reproduction. 2021.
https://doi.org/10.1016/j.immuni.2021.02.001
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.
Modified Vaccinia Ankara Vector-Based Vaccine Protects Macaques from SARS-CoV-2 Infection, Immune Pathology and Dysfunction in the Lung
Routhu et al., Immunity. 2021.
https://doi.org/10.1016/j.immuni.2021.02.001
Any SARS-CoV-2 vaccine may have limitations such as need for ultracold storage, poor induction of CD8+ T cell response, or lack of cross-reactivity with emerging strains. Thus, multiple vaccines may be needed to bring COVID-19 under control. Using rhesus macaques, researchers showed that a modified vaccinia Ankara (MVA) vector-based SARS-CoV-2 vaccine expressing prefusion-stabilized spike protein induced strong neutralizing antibody and CD8+ T cell responses. The vaccine protected macaques from SARS-CoV-2 infection as well as infection-induced inflammation and B cell abnormalities in the lung. These results are promising considering the excellent safety and performance of MVA vector-based vaccines for other pathogens. Supported by ORIP (P51OD011132, S10OD026799) and NIAID.
SARS-CoV-2 Induces Robust Germinal Center CD4 T Follicular Helper Cell Responses in Rhesus Macaques
Lakshmanappa et al., Nature Communications. 2021.
https://www.nature.com/articles/s41467-020-20642-x
SARS-CoV-2 infection in both sexes of rhesus macaques, either infused with convalescent plasma, normal plasma, or receiving no infusion, resulted in transient accumulation of pro-inflammatory monocytes and proliferating CD4 T follicular helper (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.
Severely Ill COVID-19 Patients Display Impaired Exhaustion Features in SARS-CoV-2-Reactive CD8+ T Cells
Kusnadi et al., Science Immunology. 2021.
https://immunology.sciencemag.org/content/6/55/eabe4782.long
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, and NIGMS.
A Modular Master Regulator Landscape Controls Cancer Transcriptional Identity
Paul et al., Cell. 2021.
https://www.sciencedirect.com/science/article/pii/S0092867420316172
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) and NCI.
Endogenous Zebrafish Neural Cre Drivers Generated by CRISPR/Cas9 Short Homology Directed Targeted Integration
Almeida et al., Scientific Reports. 2021.
https://pubmed.ncbi.nlm.nih.gov/33462297/
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 (R24OD020166).
Myelin‐Specific T Cells in Animals With Japanese Macaque Encephalomyelitis
Govindan et al., Annals of Clinical and Translational Neurology. 2021.
https://onlinelibrary.wiley.com/doi/10.1002/acn3.51303
Investigators characterized the CD4+ and CD8+ T cells in demyelinating Japanese macaque encephalomyelitis (JME) lesions in age‐ and sex‐matched macaques and discovered differences in expression of myelin antigen sequences in the T cell. Mapping myelin epitopes revealed a heterogeneity in T cell responses among JME animals, which are associated with a proinflammatory pathogenic role in multiple sclerosis (MS). These findings draw further parallels between JME and MS and support the hypothesis that JME and possibly MS are triggered by mechanisms involving myelin damage and not myelin epitope mimicry. Supported by ORIP (P51OD011092) and NINDS.