A genetic and immunological investigation of JAK-STAT in the pathogenesis of Celiac Disease

该提案详细说明了一项全面的五年培训计划，以扩大我作为内科科学家的技能。 研究计划的重点是JAK-STAT通路在腹腔疾病(CED)中的作用，但培训计划包括 在人类主题研究、生物信息学和基于实验室的T细胞培训中的广泛传授 免疫学。CED是一种免疫介导的胃肠道疾病，发生在患者允许的 人类白细胞抗原DQ2/人类白细胞抗原DQ8的遗传背景。唯一可用的治疗CED的方法是遵循无麸质饮食。 患有生殖系功能获得(GOF)STAT3突变和唐氏综合征的患者估计有40- CED的风险比普通人群分别高出两倍和五倍，同时易患上 其他自身免疫性疾病，包括甲状腺炎和I型糖尿病。唐氏综合征患者有 21号染色体上干扰素受体基因的三个副本，从而构成JAK-STAT 激活。中心假设是干扰素和细胞因子介导的JAK-STAT过度激活可能 导致T细胞对面筋的耐受性丧失，并增加CED的风险。这项研究的总体目标是 探讨JAK-STAT激活在CED发病机制中的作用。要做到这一点，首先，我们 将对100例家族性CED患者进行遗传学调查，并在 JAK-STAT通路。我们还将扩大CED患者的队列，这些患者患有影响 JAK-STAT激活，包括GOF-STAT3和DS患者。第二，我们将评估关键分子在 JAK-STAT途径，包括激动剂、受体、STAT及其磷酸化形式，在 单核细胞和CD4+T细胞鉴定活动性CED的分子特征。第三，我们将调查 JAK-STAT在面筋特异性CD4+T细胞上过度激活的机制 扩增T细胞库，然后测试JAK抑制剂对面筋特异性T细胞的功能影响。 这项研究的结果将描绘JAK-STAT的激活及其作为CELAC治疗靶点的潜力 疾病。此外，通过拟议的补充职业发展计划，我将获得额外的 CED遗传学和免疫学临床研究培训；高级生物信息学分析 RNA-seq；和基于实验室的面筋特异性CD4+T细胞生物学培训。在这项研究和 职业发展活动，我将由一个由Timothy Wang博士领导的团队进行指导，他是一位国际 公认的内科科学家和炎性细胞因子及其在人类胃肠道中的作用专家 疾病。我致力于以患者为导向的翻译研究的独立研究员的职业生涯； 并设计了我的培训计划，以获得所需的知识和技能，以使 使用功能遗传学方法发现治疗靶点对该领域的重大贡献 在胃肠道疾病方面。

The proposal details a comprehensive five years training program to expand my skills as a physician-scientist. The research plan is focused on JAK-STAT pathway in Celiac Disease (CeD), but the training plan includes extensive didactics in human subject research, bioinformatics and laboratory-based training on T cell immunology. CeD is an immune mediated gastrointestinal disease that arises in patients with a permissive HLA-DQ2/HLA-DQ8 genetic background. The only available treatment for CeD is to follow a gluten-free diet. Patients with germline gain-of-function (GOF) STAT3 mutations and Down Syndrome have an estimated 40- fold and 5-fold higher risk of CeD, respectively, than the general population, along with a predisposition for other autoimmune diseases, including thyroiditis and Type I Diabetes. Patients with Down Syndrome have three copies of the gene for interferon (IFN) receptors on chromosome 21 and thus constitutive JAK-STAT activation. The central hypothesis is that interferon and cytokines mediated JAK-STAT overactivation may cause loss of T cell tolerance to gluten and an increased risk for CeD. The overall goal of this research is to investigate the mechanism of JAK-STAT activation in the pathogenesis of CeD. To accomplish this, first, we will conduct genetic investigation in 100 index cases with familial CeD and search for genetic variants in the JAK-STAT pathway. We will also expand our cohort of CeD patients with known genetic diseases affecting JAK-STAT activation, include patients with GOF-STAT3 and DS. Second, we will assess the key molecules in the JAK-STAT pathway, including agonists, receptors, STAT and their phosphorylated forms, ISGs in monocyte and CD4+ T cells to identify a molecular signature of active CeD. Third, we will investigate the mechanisms of JAK-STAT overactivation on gluten-specific CD4+ T cells through both single cell RNA-Seq and amplified T cell libraries, followed by testing the functional impact of JAK inhibitors on gluten-specific T cells. The results of this study will delineate JAK-STAT activation and its potential as a therapeutic target for Celiac Disease. Furthermore, through the proposed complementary career development plan, I will gain additional training in clinical investigation on the genetics and immunology of CeD; advanced bioinformatic analysis with RNA-seq; and laboratory-based training in gluten specific CD4+ T cells biology. Throughout this research and career development activities, I will be mentored by a team lead by Dr. Timothy Wang, an internationally recognized physician-scientist and an expert in inflammatory cytokines and their role in human gastrointestinal diseases. I am committed to a career as an independent investigator in patient-oriented translational research; and have designed my training plan to acquire the knowledge and skills needed to make a meaningful and substantial contribution to the field by using a functional genetics approach to discover the therapeutic targets in gastrointestinal diseases.