The role of SH2B3 in regulating CD8 T cells in Type 1 Diabetes

项目摘要/摘要 这个项目是为埃里克·阿伦斯帕奇博士申请的一项小额赠款计划R03，他是麻省理工学院的助理教授 华盛顿大学(UW)儿科。他是美国国立卫生研究院临床指导项目的获得者 NIDDK颁发的科学家研究职业发展奖(K08)，目前处于第四年。Dr。 Allenspach是儿科风湿科和免疫科的主治医师， 对治疗儿童自身免疫性疾病的临床和研究感兴趣。Allenspach博士的特效药 研究兴趣在于了解调节自身免疫的分子机制，包括罕见的变异。 和风险等位基因。他的长期职业目标是成为一名独立资助的首席调查员，研究 在两种动物模型和使用基础和人类样本的自身免疫的遗传机制 翻译方法。 在本申请中，Allenspach博士正在申请NIH R03资金，以支持一个新的研究领域 与他的K08有关联，但又不同。Allenspach博士建议研究一种已识别的自身免疫的生物学作用 在1型糖尿病(T1D)小鼠模型中，适配蛋白SH2B3的风险变异对T细胞功能的影响。 SH2B3基因中的一个遗传等位基因(Rs3184504)与T1D有很强的相关性。在这 建议，我们利用小鼠模型来了解SH2B3功能降低如何影响T细胞。K08 提案集中在SH2B3在塑造髓系发育、功能和抗原提呈方面的作用，如 链接到T1D。现在，我们小组的初步数据已经确定了SH2B3在T细胞中的内在作用 调节IL-2受体途径。已经发现遗传风险等位基因之间存在很强的关联 (rs3184504*T)和T1D，该等位基因编码一种亚型SH2B3蛋白。S2b3基因缺乏症 小鼠CD8T细胞使其对体内耐受机制产生耐受性 糖尿病模型。驱动这种自身免疫的机制尚不清楚。 在这项提案中，我们将测试SH2B3的缺陷是否会改变TCR信令阈值 增殖分化。我们将利用一个成熟的小鼠TCR-转基因模型 用TCR信号报告，并在体外和体内进行测试。我们将探讨SH2B3是否规范了所有 常见的伽马链受体以及这个信号如何与TCR信号相交。我们还将测试 SH2B3缺乏症是否偏向末端效应记忆细胞，这是一种已知的致病表型 CD8细胞与T1D进展相关。这些基础研究将有助于了解机械论 了解SH2B3基因风险变异在促进糖尿病中的作用，并可以指导未来的人类研究。它 预计这些研究将提供制定 自主研究方向，并申请R01年终资助的职业发展支持。

PROJECT SUMMARY/ABSTRACT This project is a small grant program R03 application for Dr. Eric Allenspach, an Assistant Professor in the Department of Pediatrics at the University of Washington (UW). He is a recipient of an NIH Mentored Clinical Scientist Research Career Development Award (K08) from the NIDDK and is currently in year four. Dr. Allenspach is an attending physician in both the divisions of Pediatric Rheumatology and Immunology and has a clinical and research interest in the treatment of pediatric autoimmune conditions. Dr. Allenspach’s specific research interest is understanding the molecular mechanisms regulating autoimmunity including rare variants and risk alleles. His long-term career goal is to become an independently funded principal investigator studying genetic mechanisms of autoimmunity in both animal models and using human samples using basic and translational approaches. In the present application, Dr. Allenspach is requesting NIH R03 funding to support a new area of research related, yet distinct, to his K08. Dr. Allenspach proposes studying the biologic role of an identified autoimmune risk variant in the adaptor protein SH2B3 on the function of T cells in mouse models of type 1 diabetes (T1D). A strong association has been found between a genetic allele (rs3184504) in the SH2B3 gene and T1D. In this proposal, we utilize murine modeling to understand how reduced SH2B3 function affects T cells. The K08 proposal focused on the role of SH2B3 in shaping myeloid development, function and antigen presentation as linked to T1D. Now, preliminary data from our group has identified a clear T cell-intrinsic role for SH2B3 in regulating the IL-2 receptor pathway. A strong association has been found between the genetic risk allele (rs3184504*T) and T1D and this allele encodes for a hypomorphic SH2B3 protein. Deficiency of Sh2b3 in murine CD8 T cells renders them refractory to tolerance mechanisms in vivo as demonstrated in a murine diabetes model. The mechanism driving this autoimmunity is not clear. In this proposal, we will test whether deficiency in SH2B3 alters the TCR signaling threshold required for proliferation and differentiation. We will leverage a well-established murine TCR-transgenic models coupled with TCR signaling reporters and test this in vitro and in vivo. We will explore whether SH2B3 regulates all of the common gamma chain receptors and how this signaling intersects with TCR signaling. We will also test whether SH2B3 deficiency skews toward terminal effector memory cells, a known pathogenic phenotype of CD8 cells associated with T1D progression. These basic studies will help inform the mechanistic understanding of the SH2B3 genetic risk variant in promoting diabetes and can guide future human studies. It is anticipated these studies would provide the publications and preliminary data needed to develop an independent research direction and apply for R01 funding at the end of the career development support.