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Principal Investigator: Yiping Zhu
Organization: UNIVERSITY OF ROCHESTER
Fiscal Year: 2023
Award: $231,000
Funding agency: National Institute of Allergy and Infectious Diseases
ABSTRACT
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the causative agent of the coronavirus disease
2019 (COVID-19), has caused a global health crisis. Currently effective vaccines are available to prevent SARS-
CoV-2 infection, but our options for the treatment of SARS-CoV-2 infection remain limited. To inform the
development of innovative interventions designed to counteract SARS-CoV-2 infection, we need a better
understanding of the molecular details of SARS-CoV-2 infection, especially the host dependency factors that are
required for virus infection. We have performed a genome-wide CRISPR knockout screen and identified PCBD1,
TRAF3, and RAD54L2 as novel host factors required for SARS-CoV-2 infection. Our preliminary data showed
that PCBD1, TRAF3, and RAD54L2 are required for the transcription of angiotensin converting enzyme 2
(ACE2). ACE2 is the primary receptor for the cellular entry of SARS-CoV-2, and plays key roles in virus infection
and pathogenesis. ACE2 is differentially expressed in a wide variety of human tissues. However, the molecular
basis for the transcriptional regulation of ACE2 in different tissues remains unexplored, which constitutes a clear
knowledge gap in SARS-CoV-2 research. In Aim 1, we will determine the function of PCBD1, TRAF3, and
RAD54L2 in transcriptional regulation of ACE2 in four cell lines derived from different tissues: Vero E6 (monkey
kidney), Calu-3 (human lung), C2BBe1 (human large intestine), and HK-2 (human kidney). SARS-CoV-2
continuously evolves by genetic mutations for the adaption to the host. We identified a number of mutations in
Spike (S) and Nucleocapsid (N) that may enhance the replication of SARS-CoV-2 in cells expressed low levels
of ACE2. Some mutation (such as N S194L) was prevalent in India and Mexico during early circulation,
suggesting these variants may contribute to the viral adaption to the host. In Aim 2, we will analyze the function
of Spike (N81S, L242P, E484D, and P1079T) and Nucleocapsid (S194L) variants in virion production and cell
entry of SARS-CoV-2. Our proposed studies will provide new information regarding the transcriptional regulation
of ACE2 and the adaption of SARS-CoV-2. In the long term, these studies will provide new targets and strategies
for the development of antiviral drugs.
Terms: <2019 novel corona virus><2019 novel coronavirus><2019-nCoV><ACE2><Amino Acid Substitution><Antibodies><Architecture><Binding><Binding Sites><Body Tissues><CD40 Receptor-Associated Factor 1><CD40bp Protein><COVID-19><COVID-19 infection><COVID-19 therapy><COVID-19 treatment><COVID-19 virus><COVID-19 virus infection><COVID19><COVID19 infection><COVID19 therapy><COVID19 treatment><COVID19 virus><CRAF1 Protein><CRISPR><CRISPR/Cas system><CV-19><CV19><Cell Body><Cell Line><CellLine><Cells><Chromatin><Circulation><Clustered Regularly Interspaced Short Palindromic Repeats><CoV-2><CoV2><Combining Site><Complex><DNA Alteration><DNA Binding><DNA Binding Interaction><DNA Sequence Alteration><DNA bound><DNA mutation><Data><Data Bases><Databases><Dependence><Deposit><Deposition><Development><Engineering / Architecture><Epigenetic><Epigenetic Change><Epigenetic Mechanism><Epigenetic Process><Gene Transcription><Genetic Alteration><Genetic Change><Genetic Transcription><Genetic defect><Genetic mutation><HNF-1 protein><Host Factor><Host Factor Protein><Human><India><Integration Host Factors><Kidney><Kidney Urinary System><Knock-out><Knockout><Knowledge><LAP-1 Protein><Large Intestine><Lung><Lung Respiratory System><Maps><Mexico><Modern Man><Molecular><Molecular Interaction><Monkeys><Mutation><Nucleocapsid><Ortholog><Orthologous Gene><Play><Production><Promoter Regions><Promotor Regions><RNA Expression><Reactive Site><Receptor Cell><Receptor Protein><Regulation><Research><Residual><Residual state><SARS corona virus 2><SARS-CO-V2><SARS-COVID-2><SARS-CoV-2><SARS-CoV-2 infection><SARS-CoV-2 therapy><SARS-CoV-2 treatment><SARS-CoV2><SARS-CoV2 infection><SARS-associated corona virus 2><SARS-associated coronavirus 2><SARS-coronavirus-2><SARS-related corona virus 2><SARS-related coronavirus 2><SARSCoV2><Sequence Alteration><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-associated coronavirus 2><Severe Acute Respiratory Syndrome-related coronavirus 2><Severe acute respiratory syndrome associated corona virus 2><Severe acute respiratory syndrome coronavirus 2><Severe acute respiratory syndrome coronavirus 2 infection><Severe acute respiratory syndrome related corona virus 2><Strains Cell Lines><TNF receptor-associated factor 3><TRAF-3><TRAF3><Testing><Tissues><Transcription><Transcription Activation><Transcription Regulation><Transcriptional Activation><Transcriptional Control><Transcriptional Regulation><Variant><Variation><Viral><Viral Diseases><Viral Pathogenesis><Virion><Virus><Virus Diseases><Virus Particle><Wuhan coronavirus><access to vaccination><access to vaccines><angiotensin converting enzyme 2><angiotensin converting enzyme II><anti-viral development><anti-viral drug development><anti-viral therapeutic development><anti-viral therapy development><antiviral development><antiviral drug development><antiviral therapeutic development><antiviral therapy development><cofactor><corona virus disease 2019><coronavirus disease 2019><coronavirus disease 2019 infection><coronavirus disease 2019 therapy><coronavirus disease 2019 treatment><coronavirus disease 2019 virus><coronavirus disease-19><coronavirus disease-19 virus><coronavirus infectious disease-19><cultured cell line><data base><developing anti-viral agent><developing anti-viral drug><developing anti-viral therapeutic><developing anti-viral therapy><developing antiviral agent><developing antiviral drug><developing antiviral therapeutic><developing antiviral therapy><developmental><differential expression><differentially expressed><epigenetically><genetic promoter element><genetic promoter sequence><genome mutation><genome scale><genome-wide><genomewide><genomic alteration><global health><hCoV19><hepatic nuclear factor 1><hepatocyte nuclear factor 1><human tissue><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><innovate><innovation><innovative><intervention design><large bowel><liver specific transcription factor LF-B1><nCoV2><novel><nuclear protein LF-B1><prevent><preventing><promoter sequence><protein protein interaction><pulmonary><receptor><recruit><renal><severe acute respiratory syndrome coronavirus 2 therapy><severe acute respiratory syndrome coronavirus 2 treatment><therapeutic agent development><therapeutic development><therapy design><transcription factor APF><transcription factor HNF1><transcription factor HP1><transcription factor LFB1><transcriptional differences><treat COVID-19><treat COVID19><treat SARS-CoV-2><treat coronavirus disease 2019><treat severe acute respiratory syndrome coronavirus 2><treatment design><vaccination access><vaccination availability><vaccine access><vaccine availability><validation studies><viral infection><virus infection><virus pathogenesis><virus-induced disease>