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

• 批准号: 10642957
• 负责人: Declan McCole
• 金额: $ 37.81万
• 依托单位: UNIVERSITY OF CALIFORNIA RIVERSIDE
• 依托单位国家: 美国
• 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/10642957
• 关键词: 2019-nCoV; ACE2; 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; Collaborations; Country; Data; Diabetes Mellitus; Diarrhea; Disease; Disputes; Early Diagnosis; Electrolytes; Epithelial Cells; Epithelium; 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; Macrophage; Mediating; Membrane Glycoproteins; Molecular; Mus; National Institute of Diabetes and Digestive and Kidney Diseases; Non-Receptor Protein Tyrosine Phosphatase Gene; Oral; Organoids; Outcome; Patients; Phenotype; Play; Predisposition; Property; Protein Tyrosine Phosphatase; Proteins; Pulmonary Inflammation; RNA Viruses; Regulation; Reporting; Respiratory Disease; 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; gut inflammation; in vitro Model; in vivo; in vivo Model; inhibitor; innovation; intestinal epithelium; knock-down; loss of function; novel; pandemic disease; particle; prevent; prophylactic; receptor; risk mitigation; risk variant; severe COVID-19; synergism; transmission process; vaccine trial

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）大流行已影响220次超过7000万个人 国家（www.who.int）。 COVID-19的主要临床混杂因素是缺乏对宿主因素的了解 在某些患者中促进对SARS-COV-2感染和更严重的胃肠道症状的敏感性。 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 对多种肠道疾病的易感性的遗传标记，RS1893217功能丧失PTPN2变体， 来自炎症性肠病的肠活检中SARS-COV-2受体ACE2的水平升高 （IBD）患者。我们假设拥有PTPN2功能丧失变体的个体可能更多 易受SARS-COV-2感染。我们已经在IBD中验证了我们的初始发现 PTPN2基因活性丧失的患者导致ACE2表达增加，SARS-COV-2 使用体外和/或体内模型，尖峰蛋白细胞进入。我们的总体目标是 机械学通过上调来确定PTPN2活动丧失如何促进SARS-COV-2细胞进入 ACE2表达的表达，如果临床认可的JAK抑制剂Tofacitinib可以缓解这种敏感性。 AIM 1将解决PTPN2如何限制ACE2的表达以及其他宿主病毒进入的辅助因素，以便在功能上 限制了人类肠上皮细胞（IEC）系的病毒进入，并表达IBD风险 PTPN2变体RS1893217，以及来自PTPN2缺陷小鼠的肠to。 AIM 2将o确定PTPN2的方式 IEC中缺乏症来改变小鼠适应的SARS-COV-2以及肠道的严重程度 与腹泻症状相关的结果是Covid-19患者。 AIM 3将机械地确定JAK是否 抑制剂，tofacitib可以在体内标准化ACE2水平，以减少IEC，人类和小鼠的病毒进入 来自PTPN2生成的IBD患者的肠动物，PTPN2缺陷小鼠和细胞。我们还将确定tofacitinib是否 可以修改IBD患者的ACE2水平。我们已经建立了新颖的小鼠系和体外模型系统 这项研究。我们将在一系列创新和既定的方法中使用这些模型系统，以使我们能够 机械学上定义了SARS-COV-2与上皮之间基本相互作用的PTPN2调节 病毒进入所需的细胞并随后引起共vid-19。这些实验代表了 令人振奋的新指导，使调查团队的专业知识协同。这些研究的结果是 有望在识别识别的重大进展1）如何与几种高度相关的遗传风险变异 NIDDK疾病可以增加对SARS-COV-2感染的敏感性； 2）SARS-COV-2如何破坏上皮 诚信和稳态以促进腹泻； 3）如何将批准的IBD治疗性重新使用为 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 现有的JAK抑制剂可以针对预防性或治疗性给药，以及遗传筛查 SARS-COV-2疫苗试验的个体。