Immune Function and the Progression to Type 1 Diabetes

1型糖尿病（T1 D）是一种在自身免疫性破坏胰岛素产生后出现的疾病， 胰腺β细胞。以前的研究，包括近120同行评议的文章，支持这P01超过 目前的资助周期，已经证明，患有T1 D或T1 D风险增加的个体显示出一系列先天性 以及与>150个风险相关基因座的基因型相关的适应性免疫异常。事实上，这些努力 已经确定了一系列与T1 D相关的免疫功能障碍，这些免疫功能障碍受到遗传学的强烈影响 (e.g.,功能突变的丧失/获得、启动子/增强子变体、外显子使用的变化），并且似乎驱动 自体免疫然而，T1 D风险基因座对这些过程的复杂贡献仍不清楚。所以我们 寻求这种P01更新的目标是阐明T1 D相关变异在整个人类中的机制。 基因组（项目1），特别是（SH 2B 3 [项目2]，HLA-II区，SIRPG，CD 226 [项目3]），赋予 表型和功能性免疫缺陷。这项研究将共同测试两个总体假设- 1)T1 D风险变体的影响将因组织、细胞亚群和激活状态而异，以及2）风险变体、细胞 应激和免疫途径的缺陷是导致胰腺炎自身免疫性破坏的关键， 导致T1 D的细胞。这三个独立但高度互动的项目在共享数据方面有着悠久的历史， 使用创新技术，并评估新鲜和冷冻保存的样本， 患有T1 D或有T1 D风险的受试者，通过两个核心机构确定：核心A-管理/样本 采集和核心B-生物样本库/生物统计学/生物信息学。项目1将实施我们的新胰腺切片 培养系统和TCR重定向的“化身”来评估靶器官中炎症的影响。项目 2将使用基因型选择的UFDI研究库样品（核心B）、胰腺组织和分化的基因- 编辑iPSC以询问SH 2B 3同种异型如何影响先天免疫细胞功能，T细胞活化， 通过发炎的脉管系统运输项目3建议研究胰腺淋巴结（pLN）、脾脏和 外周血，以检验T1 D相关遗传风险变异改变TCR库的假设， 以细胞和组织限制的方式改变基因表达途径。这些建议的研究得到进一步支持 佛罗里达大学丰富的T1 D研究环境，包括胰腺 来自糖尿病胰腺器官供体网络（nPOD）的淋巴组织样本，相互作用 与主要的临床试验网络，包括T1 D TrialNet，以及与种族/遗传影响有关的特别努力， 祖先（通过高度活跃的招募工作成为可能）对T1 D异质性的影响。预期的成功 从拟议的研究应该提供：1）新的见解免疫和遗传的影响， 有助于T1 D; 2）与T1 D相关的疾病易感性和自身免疫活性的新生物标志物 疾病;和3）可以显着改善发展的前景，一种有效的治疗， 预防和/或逆转T1 D。

Type 1 diabetes (T1D) is a disorder that arises following the autoimmune destruction of insulin producing pancreatic β-cells. Previous studies, including nearly 120 peer reviewed-articles supported by this P01 over the current grant cycle, have demonstrated that individuals with or at increased-risk for T1D display a series of innate and adaptive immunological abnormalities linked to genetypes at >150 risk-associated loci. Indeed, these efforts have identified a series of immune dysfunctions associated with T1D that are strongly influenced by genetics (e.g., loss/gain of function mutations, promoter/enhancer variants, shifts in exon usage) and appear to drive autoimunity. Yet the complex contributions of T1D-risk loci to these processes remain quite unclear. Thus, our goal in seeking renewal of this P01 is to elucidate the mechanisms by which T1D-associated variants across the genome (Project 1), and specifically (SH2B3 [Project 2], HLA-II region, SIRPG, CD226 [Project 3]), impart phenotypic and functional immune defects. The research proposed will collectively test two overall hypotheses— 1) the impact of T1D-risk variants will vary by tissue, cell subset and activation state, and 2) risk variants, cellular stress, and defects in immunologic pathways are key to engender the autoimmune destruction of pancreatic - cells that results in T1D. The three separate but highly interactive Projects have a strong history of sharing data, using innovative technologies, and assessing fresh and cryopreserved samples from well characterized human subjects with or at risk for T1D, as ascertained through two Core facilities: Core A– Administrative/Sample Acquisition and Core B– Biobank/Biostatistics/Bioinformatics. Project 1 will implement our novel pancreas slice culture system and TCR-redirected “avatars” to evaluate the impact of inflammation in the target organ. Project 2 will use genotype selected UFDI Study Bank samples (Core B), pancreas tissues, and differentiated gene- edited iPSCs to interrogate how SH2B3 allotypes impact innate immune cell function, T cell activation, and trafficking through inflamed vasculature. Project 3 proposes to study pancreatic lymph node (pLN), spleen, and peripheral blood to test the hypothesis that T1D-associated genetic risk variants alter the TCR repertoire and gene expression pathways in a cell- and tissue-restricted manner. These proposed studies are further supported by the rich environment for T1D research at the University of Florida that includes resources such as pancreatic and lymphoid tissue samples from the Network for Pancreatic Organ donors with Diabetes (nPOD), interactions with major clinical trial networks including T1D TrialNet, and special efforts related to the impact of race/genetic ancestry (made possible through highly active recruiting efforts) on T1D heterogeneity. The successes expected from the proposed studies should provide: 1) novel insights into the immune and genetic influences that contribute to T1D; 2) novel biomarkers for disease susceptibility and autoimmune activity associated with the disease; and 3) could dramatically improve prospects for the development of an effective therapeutic capable of preventing and/or reversing T1D.