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Principal Investigator: WEI CHEN
Organization: UNIVERSITY OF CALIFORNIA LOS ANGELES
Fiscal Year: 2022
Award: $234,000
Funding agency: National Institute of Dental and Craniofacial Research
Abstract
The long-term goal of our research is to develop effective therapies against SARS-CoV-2-elicited disease
Covid-19. SARS-CoV-2 spike (S) protein binds to its cognate receptor, angiotensin-converting enzyme 2
(ACE2), and in concert with host proteases, principally transmembrane serine proteases (TMPRSSs),
promotes cellular entry. Co-expressions of these viral entry factors are found in the epithelial cells of
many organs, including oral mucosa. Recently, oral mucosa has been found to be a potentially high-risk
route of SARS-CoV-2 infection. GRHL2, as a transcriptional factor, modulates epithelial gene expression
and plasticity through multiple signaling pathways. ACE2 and TMPRSS2 are strongly expressed in
epithelial cells, coinciding with the pattern of GRHL2 expression in oral mucosa. GRHL2 regulates ACE2
and TMPRSS2 expression in normal human oral keratinocytes (NHOK). Ectopic expression of GRHL2
induces ACE2 and TMPRSSs expression, while knockdown of GRHL2 leads to decreased levels of these
entry factors. ACE2 is an interferon-stimulated gene. IFN-α activates ACE2 expression in part through
regulating GRHL2. Small-molecule inhibitors of GRHL2 suppress viral entry factor expression in NHOK.
ACE2 and TMPRSS2 are widely expressed in oral mucosal epithelium, knockout of GRHL2 leads to
suppression of ACE2 and TMPRSS2 expression in vivo. These findings indicate that GRHL2 is required
for ACE2 and TMPRSS expression in NHOK cultured in vitro and oral mucosal epithelium in vivo. Based
on these data, we hypothesize that targeting GRHL2 inhibits SARS-CoV-2 transmission into oral
mucosal epithelium through regulating viral entry factor expression. This novel hypothesis will be
tested by examining the molecular mechanism underlying GRHL2 regulates expression of SARS-CoV-2
entry factors (ACE2 and TMPRSSs) in oral mucosal epithelial cells. Analysis of variance methods will be
used to compare means of these gene expression levels. We will also generate a pseudotyped virus in
which vesicular stomatitis virus (VSV) green fluorescent protein (GFP) reporter virus expressing SARS-
CoV-2 S in replacement of native glycoprotein. Utilizing this chimeric virus, a high throughput assay will
be developed to screen GRHL2 inhibitors, which possess the potentials to abrogate SARS-CoV-2 virus
infection into oral mucosal epithelium.
Terms: <(IFN) α><(IFN)-α><(IFN)α><2019 novel corona virus><2019 novel coronavirus><2019-nCoV><2019-nCoV S protein><2019-nCoV spike glycoprotein><2019-nCoV spike protein><3-D><3-Dimensional><3D><ACE2><ANOVA><Alferon><Analysis of Variance><Assay><Basal Transcription Factor><Basal transcription factor genes><Binding><Binding Proteins><Bioassay><Biologic Assays><Biological Assay><Body Tissues><Buccal Cavity><Buccal Cavity Head and Neck><Buccal Mucosa><COVID><COVID crisis><COVID epidemic><COVID infected patient><COVID pandemic><COVID patient><COVID positive patient><COVID-19><COVID-19 S protein><COVID-19 crisis><COVID-19 epidemic><COVID-19 global health crisis><COVID-19 global pandemic><COVID-19 health crisis><COVID-19 infected patient><COVID-19 infection><COVID-19 pandemic><COVID-19 patient><COVID-19 positive patient><COVID-19 public health crisis><COVID-19 spike glycoprotein><COVID-19 spike protein><COVID-19 transmission><COVID-19 virus><COVID-19 virus transmission><COVID19><COVID19 S protein><COVID19 crisis><COVID19 epidemic><COVID19 global health crisis><COVID19 global pandemic><COVID19 health crisis><COVID19 infection><COVID19 pandemic><COVID19 patient><COVID19 positive patient><COVID19 public health crisis><COVID19 spike glycoprotein><COVID19 spike protein><COVID19 virus><CRISPR approach><CRISPR based approach><CRISPR method><CRISPR methodology><CRISPR technique><CRISPR technology><CRISPR tools><CRISPR-CAS-9><CRISPR-based method><CRISPR-based technique><CRISPR-based technology><CRISPR-based tool><CRISPR/CAS approach><CRISPR/Cas method><CRISPR/Cas technology><CRISPR/Cas9><CRISPR/Cas9 technology><CV-19><CV19><Cas nuclease technology><Cavitas Oris><Cell Body><Cell Differentiation><Cell Differentiation process><Cell Growth in Number><Cell Multiplication><Cell Proliferation><Cell membrane><Cells><Cellular Proliferation><Clustered Regularly Interspaced Short Palindromic Repeats approach><Clustered Regularly Interspaced Short Palindromic Repeats method><Clustered Regularly Interspaced Short Palindromic Repeats methodology><Clustered Regularly Interspaced Short Palindromic Repeats technique><Clustered Regularly Interspaced Short Palindromic Repeats technology><CoV disease><CoV-2><CoV2><Containment><Cytoplasmic Membrane><Data><Dysfunction><Ectopic Expression><Epigenetic><Epigenetic Change><Epigenetic Mechanism><Epigenetic Process><Epithelial><Epithelial Cells><Epitheliasin Gene><Esteroproteases><Foundations><Functional disorder><Gene Expression><Gene Transcription><General Transcription Factor Gene><General Transcription Factors><Genes><Genetic Transcription><Gingiva><Gingival><Glycoproteins><Goals><Green Fluorescent Proteins><Health><High Throughput Assay><Human><IFN><IFN Alpha><IFN α><IFN-α><IFNa><IFNα><In Vitro><Infection><Interferon Alfa-n3><Interferon-alpha><Interferon-α><Interferons><Knock-out><Knockout><Leukocyte Interferon><Ligand Binding Protein><Ligand Binding Protein Gene><Light><Lymphoblast Interferon><Lymphoblastoid Interferon><Mediating><Methods><Mice><Mice Mammals><Modeling><Modern Man><Molecular><Molecular Interaction><Mouth><Mouth Mucosa><Murine><Mus><Oral><Oral Mucosa><Oral cavity><Oral mucous membrane structure><Organ><Outcome><PRSS10><Pattern><Peptidases><Peptide Hydrolases><Photoradiation><Physiopathology><Plasma Membrane><Play><Predisposition><Process><Protease Gene><Proteases><Protein Binding><Proteinases><Proteolytic Enzymes><Public Health><Publishing><RNA Expression><Receptor Protein><Reporter><Research><Role><Route><SARS corona virus 2><SARS-CO-V2><SARS-COVID-2><SARS-CoV-2><SARS-CoV-2 S protein><SARS-CoV-2 epidemic><SARS-CoV-2 global health crisis><SARS-CoV-2 global pandemic><SARS-CoV-2 infected patient><SARS-CoV-2 infection><SARS-CoV-2 inhibitor><SARS-CoV-2 pandemic><SARS-CoV-2 patient><SARS-CoV-2 positive patient><SARS-CoV-2 spike glycoprotein><SARS-CoV-2 spike protein><SARS-CoV-2 transmission><SARS-CoV2><SARS-CoV2 S protein><SARS-CoV2 epidemic><SARS-CoV2 infection><SARS-CoV2 pandemic><SARS-CoV2 spike glycoprotein><SARS-CoV2 spike protein><SARS-associated corona virus 2><SARS-associated coronavirus 2><SARS-coronavirus-2><SARS-coronavirus-2 epidemic><SARS-coronavirus-2 pandemic><SARS-related corona virus 2><SARS-related coronavirus 2><SARSCoV2><Salivary Glands><Salivary Glands Head and Neck><Serine Endopeptidases><Serine Protease><Serine Protein Hydrolases><Serine Proteinases><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 CoV 2 epidemic><Severe Acute Respiratory Syndrome CoV 2 pandemic><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 S protein><Severe acute respiratory syndrome coronavirus 2 epidemic><Severe acute respiratory syndrome coronavirus 2 infection><Severe acute respiratory syndrome coronavirus 2 inhibitor><Severe acute respiratory syndrome coronavirus 2 pandemic><Severe acute respiratory syndrome coronavirus 2 spike glycoprotein><Severe acute respiratory syndrome coronavirus 2 spike protein><Severe acute respiratory syndrome related corona virus 2><Signal Pathway><Site><Susceptibility><TMPRSS2><TMPRSS2 gene><Testing><Tissues><Transcription><Transcription Factor Proto-Oncogene><Transcription Regulation><Transcription factor genes><Transcriptional Control><Transcriptional Regulation><Transduction Gene><Tropism><VSV><Variance Analyses><Vector Mediated Transfer Genes><Vesicular Stomatitis Virus><Vesicular stomatitis Indiana virus><Viral><Viral Diseases><Virus><Virus Diseases><Wuhan coronavirus><angiotensin converting enzyme 2><angiotensin converting enzyme II><base><block SARS-CoV-2><block severe acute respiratory syndrome coronavirus 2><block viral entry><bound protein><corona virus disease><corona virus disease 2019><corona virus disease 2019 epidemic><corona virus disease 2019 pandemic><coronavirus disease><coronavirus disease 2019><coronavirus disease 2019 S protein><coronavirus disease 2019 crisis><coronavirus disease 2019 epidemic><coronavirus disease 2019 global health crisis><coronavirus disease 2019 global pandemic><coronavirus disease 2019 health crisis><coronavirus disease 2019 infected patient><coronavirus disease 2019 infection><coronavirus disease 2019 pandemic><coronavirus disease 2019 patient><coronavirus disease 2019 positive patient><coronavirus disease 2019 public health crisis><coronavirus disease 2019 spike glycoprotein><coronavirus disease 2019 spike protein><coronavirus disease 2019 transmission><coronavirus disease 2019 virus><coronavirus disease 2019 virus transmission><coronavirus disease crisis><coronavirus disease epidemic><coronavirus disease infected patient><coronavirus disease pandemic><coronavirus disease patient><coronavirus disease positive patient><coronavirus disease-19><coronavirus disease-19 global pandemic><coronavirus disease-19 pandemic><coronavirus disease-19 patient><coronavirus disease-19 virus><coronavirus infectious disease-19><coronavirus patient><effective therapy><effective treatment><experiment><experimental research><experimental study><gain of function><hCoV19><high risk><high throughput screening><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><inhibit SARS-CoV-2><inhibit severe acute respiratory syndrome coronavirus 2><inhibit viral entry><inhibitor><keratinocyte><knock-down><knockdown><knockout gene><monolayer><mouse model><murine model><nCoV2><new drug target><new druggable target><new pharmacotherapy target><new therapeutic target><new therapy target><novel><novel drug target><novel druggable target><novel pharmacotherapy target><novel therapeutic target><novel therapy target><oral cavity epithelium><oral epithelia><oral epithelium><oral mucosae><oral mucosal><pathophysiology><patient infected with COVID><patient infected with COVID-19><patient infected with SARS-CoV-2><patient infected with coronavirus disease><patient infected with coronavirus disease 2019><patient infected with severe acute respiratory syndrome coronavirus 2><patient with COVID><patient with COVID-19><patient with COVID19><patient with SARS-CoV-2><patient with coronavirus disease><patient with coronavirus disease 2019><patient with severe acute respiratory distress syndrome coronavirus 2><plasmalemma><promoter><promotor><receptor><screening><severe acute respiratory syndrome coronavirus 2 global health crisis><severe acute respiratory syndrome coronavirus 2 global pandemic><severe acute respiratory syndrome coronavirus 2 infected patient><severe acute respiratory syndrome coronavirus 2 patient><severe acute respiratory syndrome coronavirus 2 positive patient><severe acute respiratory syndrome coronavirus 2 transmission><small molecule inhibitor><social role><three dimensional><transcription factor><translational potential><transmitted COVID-19><transmitted SARS-CoV-2><transmitted coronavirus disease 2019><transmitted severe acute respiratory syndrome coronavirus 2><viral entry blocker><viral entry inhibitor><viral infection><virus infection><virus-induced disease>