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Targeting Grainyhead-Like 2 Suppresses Entry Factors of SARS-CoV-2 in Epithelial Cells of Oral Mucosa.

靶向 Grainyhead-Like 2 可抑制口腔粘膜上皮细胞中 SARS-CoV-2 的进入因子。

基本信息

批准号：
10453095
负责人：
WEI CHEN
金额：
$ 23.4万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-04-01 至 2024-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10453095
关键词：
2019-nCoV 3-Dimensional ACE2 Analysis of Variance Binding Binding Proteins Biological Assay COVID-19 COVID-19 pandemic COVID-19 patient CRISPR/Cas technology Cell Differentiation process Cell Proliferation Cell membrane Cells Containment Data Ectopic Expression Epigenetic Process Epithelial Epithelial Cells Foundations Functional disorder Gene Expression Genes Genetic Transcription Gingiva Glycoproteins Goals Green Fluorescent Proteins Health Human In Vitro Infection Interferon-alpha Interferons Knock-out Light Mediating Methods Modeling Molecular Mus Oral Oral cavity Oral mucous membrane structure Organ Outcome Pattern Peptide Hydrolases Play Predisposition Process Public Health Publishing Reporter Research Role Route SARS-CoV-2 infection SARS-CoV-2 inhibitor SARS-CoV-2 spike protein SARS-CoV-2 transmission Salivary Glands Serine Protease Signal Pathway Site TMPRSS2 gene Testing Tissues Transcriptional Regulation Transduction Gene Tropism Vesicular stomatitis Indiana virus Viral Virus Virus Diseases base coronavirus disease effective therapy experimental study gain of function high risk high throughput screening in vivo inhibitor keratinocyte knock-down knockout gene monolayer mouse model new therapeutic target novel oral cavity epithelium promoter receptor screening small molecule inhibitor transcription factor translational potential viral entry inhibitor

项目摘要

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.

摘要