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Principal Investigator: Philip Murphy
Organization: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
Fiscal Year: 2022
Award: $15,314
Funding agency: National Institute of Allergy and Infectious Diseases
We have written an Animal Study Proposal that has been approved by the NIAID Animal Care and Use Committee and the SVC. We began research in fall, 2020 after the BSL3 and ABSL3 facilities opened for this purpose. To date, WT and huACE2 transgenic mice on a C57BL/6 background have been infected with clinical isolates of SARS-CoV-2. The huACE2 mice are highly susceptible to fatal infection whereas the WT mice survive. We are crossing mouse strains to test chemokine receptor knockout mice on the huACE2 background prioritizing receptors encoded by genes in the CCR cluster since human GWAS studies of Covid disease have reported a strong signal in this region most proximal to CCR9. Our goal is to test a WT LD100 dose first, screening for receptor knockouts that increase survival, since any positive results will have translational potential. Receptors that screen negative will be rescreened at an LDzero looking for protective chemokine receptors in the model. To date we have identified Ccr6 knockouts as having increased susceptibility to weight loss and fatal outcome. We are also attempting to develop a AAV-huACE2 transient expression system to accelerate screening by obviating the huACE2 transgenic/chemokine receptor ko crosses. We are also pursuing a strain of SARS-CoV-2 that has been adapted to mouse infection towards accelerating discovery. For each type of experiment four virus doses will be tested to determine if models for asymptomatic, mild virulence or higher virulence can be developed for each virus. Both females and males will be tested and are expected to have differences in clinical progression of coronaviral disease and histopathology, based on other infectious disease models in mice, and differences between the sexes observed in humans infected with SARS-CoV and SARS-CoV-2. In all subsequent experiments, clinical disease progression will be assessed by virus recovery from throat swabs, blood and stool; serum antibody titers, hemograms, blood chemistries and serum cytokines before and at multiple intervals after infection; as well as physical appearance, weight loss, and progression to moribundity and survival for a month after infection. Animals will be euthanized by CO2 and cervical dislocation at the latest one-month post-infection. In our experience, to observe significant differences in survival we will need 20 mice/cohort in each experiment and repeat the experiment 3 times due to the variance in survival between experiments. These numbers should be sufficient to observe significant differences in the other clinical data as well. Fewer non-infected mice will be needed since there is expected to be less variance among these controls. In this reporting period, the results of our ko screen has revealed protective effects of Ackr1 deficiency and harmful effects of Ccr2, Ccr5 and Cxcl10 deficiency.
A secondary goal is to characterize the local immune response in cultured human lung explants infected with SARS-CoV-2. We have succeeded in collaboration with L Margolis from NICHD in establishing this system and demonstrating productive infection in epithelial cells.
We have not yet published these results.
Terms: <2019 novel corona virus><2019 novel coronavirus><2019-nCoV><Animal Care and Use Committees><Animals><Antibodies><Antibody titer measurement><Appearance><Blood><Blood Chemical Analyses><Blood Chemical Analysis><Blood Reticuloendothelial System><Blood Serum><Body Weight decreased><C-C CKR-9><CC-CKR-9><CCR><CCR-9><CCR9><CCR9 gene><CO2><COVID><COVID-19><COVID-19 immune response><COVID-19 infection><COVID-19 virus><COVID19><COVID19 immune response><COVID19 infection><COVID19 virus><CV-19><CV19><Carbon Dioxide><Carbonic Anhydride><Cell Communication and Signaling><Cell Signaling><Cervical><Chemokine Receptor Gene><Clinical><Clinical Data><CoV disease><CoV-2><CoV2><Collaborations><Differences between sexes><Differs between sexes><Disease><Disease Progression><Dislocations><Disorder><Dose><Drugs><Epithelial Cells><Fatal Outcome><Feces><Female><GPR-9-6><GWA study><GWAS><Genes><Goals><Histopathology><Human><Immune response><Immunodeficiency Disorder><Immunodeficiency Syndrome><Immunologic Deficiency Syndromes><Immunological Deficiency Syndromes><Immunological response><Immunomodulation><Individual><Infection><Intracellular Communication and Signaling><KO mice><Knock-out><Knock-out Mice><Knockout><Knockout Mice><Lung><Lung Respiratory System><Medication><Mice><Mice Mammals><Modeling><Modern Man><Mouse Strains><Murine><Mus><NIAID><NICHD><National Institute of Allergy and Infectious Disease><National Institute of Child Health and Human Development><National Institute of Children's Health and Human Development><Null Mouse><Patients><Pharmaceutic Preparations><Pharmaceutical Preparations><Pharyngeal structure><Pharynx><Play><Predisposition><Publishing><Receptor Protein><Recovery><Reporting><Research><Role><SARS Virus><SARS corona virus><SARS corona virus 2><SARS coronavirus><SARS-Associated Coronavirus><SARS-CO-V2><SARS-COVID-2><SARS-CoV><SARS-CoV-1><SARS-CoV-2><SARS-CoV-2 immune response><SARS-CoV-2 infection><SARS-CoV2><SARS-CoV2 infection><SARS-Related Coronavirus><SARS-associated corona virus 2><SARS-associated coronavirus 2><SARS-coronavirus-2><SARS-related corona virus 2><SARS-related coronavirus 2><SARSCoV2><Serum><Severe Acute Respiratory Coronavirus><Severe Acute Respiratory Coronavirus 2><Severe Acute Respiratory Distress Syndrome CoV 2><Severe Acute Respiratory Distress Syndrome Corona Virus 2><Severe Acute Respiratory Distress Syndrome Coronavirus 2><Severe Acute Respiratory Syndrome CoV 2><Severe Acute Respiratory Syndrome Virus><Severe Acute Respiratory Syndrome corona virus><Severe Acute Respiratory Syndrome coronavirus><Severe Acute Respiratory Syndrome-associated coronavirus 2><Severe Acute Respiratory Syndrome-related coronavirus 2><Severe acute respiratory syndrome associated corona virus 2><Severe acute respiratory syndrome corona virus 2><Severe acute respiratory syndrome coronavirus 2><Severe acute respiratory syndrome coronavirus 2 immune response><Severe acute respiratory syndrome coronavirus 2 infection><Severe acute respiratory syndrome related corona virus 2><Sex Differences><Sexual differences><Signal Transduction><Signal Transduction Systems><Signaling><Site><Susceptibility><Swab><System><Techniques><Testing><Throat><Time><Transgenic Mice><Transgenic Organisms><Viral Pathogenesis><Virulence><Virus><Weight Loss><Weight Reduction><Wuhan coronavirus><antibody titering><base><biological signal transduction><blood chemistry><body weight loss><chemokine receptor><cohort><corona virus disease><corona virus disease 2019><coronavirus disease><coronavirus disease 2019><coronavirus disease 2019 immune response><coronavirus disease 2019 infection><coronavirus disease 2019 virus><coronavirus disease-19><coronavirus disease-19 virus><coronavirus infectious disease-19><cytokine><drug development><drug/agent><experience><experiment><experimental research><experimental study><falls><genome wide association><genome wide association scan><genome wide association studies><genome wide association study><genomewide association scan><genomewide association studies><genomewide association study><hCoV19><host response><hypoimmunity><immune deficiency disorder><immune modulation><immune regulation><immune system response><immunodeficiency><immunologic reactivity control><immunomodulatory><immunoregulation><immunoregulatory><immunoresponse><in vivo><infected with COVID-19><infected with COVID19><infected with SARS-CoV-2><infected with SARS-CoV2><infected with coronavirus disease 2019><infected with severe acute respiratory syndrome coronavirus 2><infectious disease model><insight><male><mouse model><murine model><nCoV2><new drug target><new druggable target><new pharmacotherapy target><new therapeutic target><new therapy target><novel drug target><novel druggable target><novel pharmacotherapy target><novel therapeutic target><novel therapy target><protective effect><pulmonary><receptor><screening><severe acute respiratory syndrome-CoV><sex-dependent differences><sex-related differences><sex-specific differences><small molecule><social role><stool><transgenic><translational potential><virus pathogenesis><whole genome association analysis><whole genome association studies><whole genome association study><wt-loss>