Project 2

项目摘要/摘要 1型糖尿病(T1D)的发展依赖于遗传疾病的基因之间的相互作用 易感性和环境因素被认为是导致异常免疫激活和信号传递的因素 导致耐受性的崩溃。这些复杂的相互作用对发展和 免疫细胞的功能，介导产生胰岛素的胰腺β的自身免疫破坏- 细胞。除了人类白细胞抗原(HL A)区域外，已发现50多个遗传位点 会给T1D带来不同程度的风险。然而，这些变体改变 免疫系统内部的发育和信号事件仍然没有得到很好的描述。我们的 研究旨在通过对外周血和组织的调查来解决这些知识差距 来自一组不同的研究对象(核心B)。通过这些调查，我们希望获得一个 更好地理解遗传风险变异如何影响对环境刺激的反应(例如，IFN1； 项目1)导致在自然病史和临床中观察到的高度异质性 这种疾病的表现。我们将检验一种组合易感基因的假设 变异和异常的外源性生长、存活和分化因子导致T细胞丧失 在T1D背景下的容忍度。我们建议进行一系列的目标：1)识别与T1D相关的 T1D患者及其亲属的横断面队列中的细胞和表型特征 疾病的风险程度，并通过Flow使用广泛的人类免疫表型(HIP)进行对照 细胞学结合全基因组974K单核苷酸多态(SNPs)基因分型和 复杂的生物信息学分析(与项目3)；2)确定表型和功能影响 遗传风险变异和年龄相关的生长因子对调节性和效应性T细胞功能的影响，以及3)创建 具有已知反应性和同基因细胞系统的T细胞受体(TCR)“阿凡达”，可发展为“一种疾病”。 DISH“模型用于评估抗原特异性T细胞功能。这些后一项研究将采用新颖的 工具包括慢病毒表达系统、人类T细胞的定向基因编辑和利用 独特的临床资源，以应对人类T1D上位性基因相互作用的挑战。在……里面 总而言之，来自这些目标的数据有望提供关于复杂相互作用的基本信息 影响自身免疫的遗传风险、免疫信号事件和免疫细胞发育之间的关系 疾病发病机制。这些创新平台的开发和使用将加快我们的能力 识别和测试新的治疗干预措施，以阻止T1D中β细胞的自身免疫破坏。

PROJECT SUMMARY/ABSTRACT The development of type 1 diabetes (T1D) relies on interactions between genes imparting disease susceptibility and environmental factors thought to contribute to aberrant immune activation and signaling leading to a breakdown in tolerance. These complex interactions adversely affect the development and function of the immune cells, which mediate the autoimmune destruction of insulin-producing pancreatic β- cells. In addition to the human leukocyte antigen (HLA) region, over 50 genetic loci have been identified that confer varying degrees of risk for T1D. However, the specific mechanisms by which these variants alter the development and signaling events within in the immune system remain poorly characterized. Our studies are designed to address these knowledge gaps through investigation of peripheral blood and tissues derived from a diverse cohort of study subjects (Core B). Through these investigations, we hope to gain a better understanding for how genetic risk variants influence responses to environmental stimuli (e.g., IFN1; Project 1) that lead to a large degree of heterogeneity observed in the natural history and clinical manifestations of the disease. We will test the hypothesis that a combination of susceptibility gene variants and aberrant extrinsic growth, survival, and differentiation factors lead to a loss of T cell tolerance in the context of T1D. We propose to conduct a series of Aims to: 1) identify the T1D-associated cellular and phenotypic signatures in a cross-sectional cohort of subjects with T1D, their relatives at varying degrees of risk for the disease, and controls using extensive human immunophenotyping (HIP) by flow cytometry together with genome-wide genotyping of >974K single nucleotide polymorphisms (SNPs) and sophisticated bioinformatics analyses (with Project 3); 2) determine the phenotypic and functional impact of genetic risk variants and age-associated growth factors on regulatory and effector T cell function, and 3) create T cell receptor (TCR) “avatars” with known reactivities and isogenic cellular systems to develop a “disease in a dish” model for assessing antigen-specific T cell function. These latter studies will employ novel tools including lentiviral expression systems, directed gene editing of human T cells, and the utilization of unique clinical resources to address the challenge of epistatic gene interactions in humans with T1D. In sum, data from these Aims are expected to provide essential information about the complex interactions between genetic risk, immune signaling events, and immune cell development that impact autoimmune disease pathogenesis. The development and utilization of these innovative platforms will expedite our ability to identify and test novel therapeutic interventions to halt the autoimmune destruction of β-cells in T1D.