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

该提案详细介绍了一个全面的五年培训计划，以扩大我作为一名医生科学家的技能。 该研究计划的重点是JAK-STAT途径在乳糜泻（CeD），但培训计划包括 在人类受试者研究、生物信息学和T细胞实验室培训方面的广泛教学 免疫学CeD是一种免疫介导的胃肠道疾病， HLA-DQ 2/HLA-DQ 8遗传背景。CeD唯一可用的治疗方法是遵循无麸质饮食。 具有生殖系功能获得性（GOF）STAT 3突变和唐氏综合征的患者估计有40- CeD的风险分别是一般人群的1倍和5倍， 其他自身免疫性疾病，包括甲状腺炎和I型糖尿病。唐氏综合症患者 21号染色体上干扰素（IFN）受体基因的三个拷贝，因此是组成型JAK-STAT activation.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细胞对麸质的耐受性丧失，并增加CeD的风险。本研究的总体目标是 探讨JAK-STAT激活在CeD发病中的作用机制。为此，首先，我们 将对100例家族性CeD索引病例进行遗传调查，并在 JAK-STAT途径。我们还将扩大我们的CeD患者队列，这些患者患有已知的遗传疾病， JAK-STAT激活，包括GOF-STAT 3和DS患者。其次，我们将评估 JAK-STAT途径，包括激动剂、受体、STAT及其磷酸化形式、ISG， 单核细胞和CD 4 + T细胞来鉴定活性CeD的分子特征。第三，我们将调查 JAK-STAT在谷蛋白特异性CD 4 + T细胞上过度活化的机制通过单细胞RNA-Seq和 扩增的T细胞文库，然后测试JAK抑制剂对谷蛋白特异性T细胞的功能影响。 本研究的结果将描述JAK-STAT激活及其作为乳糜泻治疗靶点的潜力。 疾病此外，通过拟议的补充职业发展计划，我将获得更多的 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;以及基于实验室的谷蛋白特异性CD 4 + T细胞生物学培训。在整个研究过程中， 职业发展活动，我将由Timothy Wang博士领导的团队指导， 他是公认的医生-科学家，也是炎症细胞因子及其在人类胃肠道疾病中作用的专家。 疾病我致力于作为一个独立的研究者在以病人为导向的转化研究的职业生涯; 并设计了我的培训计划，以获得所需的知识和技能，使一个有意义的， 通过使用功能遗传学方法来发现治疗靶点， in gastrointestinal胃肠diseases疾病.

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.