Mechanistic Characterization of the IBD Risk Gene, PTPN2, as a Novel Susceptibility Marker for Increased SARS-CoV-2 Infection

IBD 风险基因 PTPN2 作为 SARS-CoV-2 感染增加的新型易感性标记的机制表征

基本信息

批准号：
10319220
负责人：
Declan McCole
金额：
$ 37.89万
依托单位：
UNIVERSITY OF CALIFORNIA RIVERSIDE
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-08-01 至 2024-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10319220
关键词：
2019-nCoV ACE2 Address Affect Anorexia Autoimmune Diseases Awareness Biological Models Biology Biopsy COVID-19 COVID-19 pandemic COVID-19 patient COVID-19 severity COVID-19 susceptibility COVID-19 vaccine Capsid Proteins Celiac Disease Cell Line Cells Clinical Country Data Diabetes Mellitus Diarrhea Disease Disputes Early Diagnosis Electrolytes Epithelial Epithelial Cells Event Exhibits Feces Gastrointestinal tract structure Genes Genetic Markers Genetic Predisposition to Disease Genetic Risk Genetic Screening Genotype Goals High Prevalence Homeostasis Human In Vitro Individual Infection Inflammatory Bowel Diseases Inhalation Insulin-Dependent Diabetes Mellitus Integration Host Factors Intervention Intestinal Diseases Intestines Knowledge Lung Inflammation Lung diseases Mediating Membrane Glycoproteins Molecular Mus National Institute of Diabetes and Digestive and Kidney Diseases Oral Outcome Patients Phenotype Play Predisposition Property Protein Tyrosine Phosphatase Proteins RNA Viruses Regulation Reporting Respiratory Signs and Symptoms Respiratory System Risk Role Route SARS-CoV-2 infection SARS-CoV-2 spike protein Series Serum Severities Signal Transduction Susceptibility Gene Symptoms Therapeutic Ulcerative Colitis Up-Regulation Variant Viral Viral Physiology Viral reservoir Virus Virus Diseases Virus Replication Virus Shedding Vomiting aerosolized associated symptom cofactor disorder risk disorder subtype experimental study gastrointestinal symptom genetic variant in vitro Model in vivo in vivo Model inflammatory disease of the intestine inhibitor/antagonist innovation intestinal epithelium knock-down loss of function macrophage novel pandemic disease particle prevent prophylactic receptor risk variant severe COVID-19 transmission process vaccine trial

项目摘要

SUMMARY/ABSTRACT The global coronavirus disease 2019 (COVID-19) pandemic has affected over 70 million individuals in 220 countries (www.who.int). A major clinical confounder of COVID-19 is the lack of knowledge of host factors that promote susceptibility to SARS-CoV-2 infection and more severe gastrointestinal symptoms in some patients. Moreover, there is also a lack of interventions to mitigate these risks. Our preliminary studies identified that a genetic marker of susceptibility to several intestinal diseases, the rs1893217 loss-of-function PTPN2 variant, increased levels of the SARS-CoV-2 receptor, ACE2, in intestinal biopsies from inflammatory bowel disease (IBD) patients. We hypothesize that individuals harboring PTPN2 loss-of-function variants may be more susceptible to SARS-CoV-2 infection. We have already functionally validated our initial findings in IBD patients that loss of activity of the PTPN2 gene results in increased ACE2 expression and SARS-Cov-2 spike protein cellular entry, using in vitro and/or in vivo models. Our overall objective will be to mechanistically determine how PTPN2 loss-of-activity promotes SARS-CoV-2 cellular entry through upregulation of ACE2 expression, and if this susceptibility can be mitigated by the clinically approved JAK inhibitor, tofacitinib. Aim 1 will address how PTPN2 restricts expression of ACE2, and other host virus entry co-factors, to functionally restrict virus entry in human intestinal epithelial cell (IEC) lines with reduced PTPN2 or expressing the IBD risk PTPN2 variant rs1893217, as well as enteroids from Ptpn2-deficient mice. Aim 2 will o determine how PTPN2 deficiency in IECs alters the severity of infection with a mouse-adapted SARS-CoV-2, as well as intestinal outcomes relevant to diarrheal symptoms in COVID-19 patients. Aim 3 will mechanistically determine if the JAK inhibitor, tofacitinib can normalize ACE2 levels in vitro, in vivo, to reduce virus entry in IEC, human and mouse enteroids, Ptpn2-deficient mice, and cells from PTPN2-genotyped IBD patients. We will also identify if tofacitinib can modify ACE2 levels in IBD patients. We have established novel mouse lines and in vitro model systems for this study. We will use these model systems in a series of innovative and established approaches, to allow us to mechanistically define PTPN2 regulation of the fundamental interactions between SARS-CoV-2 and epithelial cells that are required for virus entry and subsequently cause COVID-19. These experiments represent an exciting new direction that synergizes the expertise of the investigative team. The results from these studies are poised to generate significant advances in identifying 1) how a genetic risk variant of high relevance to several NIDDK diseases can increase susceptibility to SARS-CoV-2 infection; 2) how SARS-CoV-2 can disrupt epithelial integrity and homeostasis to promote diarrhea; 3) how an approved IBD therapeutic can be repurposed to mitigate these risks. Therefore, the clinical implications of this study include identifying a novel mechanism that can be targeted by existing JAK inhibitors for prophylactic or therapeutic administration, and genetic screening of individuals for SARS-CoV-2 vaccine trials.

摘要/摘要 2019 年全球冠状病毒病 (COVID-19) 大流行已在 220 年影响了超过 7000 万人国家（www.who.int）。 COVID-19 的一个主要临床混杂因素是缺乏对宿主因素的了解促进某些患者对 SARS-CoV-2 感染的易感性和更严重的胃肠道症状。此外，还缺乏减轻这些风险的干预措施。我们的初步研究表明对多种肠道疾病易感性的遗传标记，rs1893217 功能丧失的 PTPN2 变体，炎症性肠病肠道活检中 SARS-CoV-2 受体 ACE2 水平升高（炎症性肠病）患者。我们假设携带 PTPN2 功能丧失变异的个体可能更容易易受 SARS-CoV-2 感染。我们已经在功能上验证了 IBD 的初步发现 PTPN2 基因活性丧失导致 ACE2 表达和 SARS-Cov-2 增加的患者使用体外和/或体内模型，刺突蛋白进入细胞。我们的总体目标是从机制上确定 PTPN2 活性丧失如何通过上调促进 SARS-CoV-2 细胞进入 ACE2 表达的影响，以及临床批准的 JAK 抑制剂托法替尼是否可以减轻这种敏感性。目标 1 将解决 PTPN2 如何限制 ACE2 和其他宿主病毒进入辅助因子的表达，以发挥功能限制病毒进入 PTPN2 降低或表达 IBD 风险的人肠上皮细胞 (IEC) 系 PTPN2 变体 rs1893217，以及来自 Ptpn2 缺陷小鼠的肠样蛋白。目标 2 将确定 PTPN2 如何 IEC 的缺乏会改变小鼠适应的 SARS-CoV-2 以及肠道细菌感染的严重程度与 COVID-19 患者腹泻症状相关的结果。目标 3 将机械地确定 JAK 是否抑制剂托法替布可以在体外、体内使 ACE2 水平正常化，以减少病毒进入 IEC、人类和小鼠体内肠类、Ptpn2 缺陷小鼠和来自 PTPN2 基因型 IBD 患者的细胞。我们还将确定托法替尼是否可以改变 IBD 患者的 ACE2 水平。我们已经建立了新的小鼠品系和体外模型系统这项研究。我们将在一系列创新和成熟的方法中使用这些模型系统，使我们能够从机制上定义 PTPN2 对 SARS-CoV-2 和上皮细胞之间基本相互作用的调节病毒进入并随后导致 COVID-19 所需的细胞。这些实验代表了令人兴奋的新方向，可以协同调查团队的专业知识。这些研究的结果是有望在识别以下方面取得重大进展：1）遗传风险变异如何与多种因素高度相关 NIDDK 疾病会增加对 SARS-CoV-2 感染的易感性； 2) SARS-CoV-2 如何破坏上皮细胞促进腹泻的完整性和体内平衡； 3) 如何将已批准的 IBD 治疗药物重新用于减轻这些风险。因此，这项研究的临床意义包括确定一种新的机制，可以通过现有的 JAK 抑制剂作为预防性或治疗性给药以及基因筛查的目标进行 SARS-CoV-2 疫苗试验的个体。