An integrated strategy to define the functional and synergistic impact of T1D causal variants

摘要 1型糖尿病（T1D）是一种复杂的疾病，由于遗传背景和环境因素， 暴露。其发病机制涉及多种生化和发育程序。而 罕见的单基因缺陷可导致T1D，我们假设T1D通常是通过以下因素的组合而产生的： 两个或多个关键免疫“程序”中的协同缺陷。可以使用与T1D相关的遗传变异 作为一个指南，以了解如何在这些程序单独和组合的变化有助于 T1D的发生和发展。尽管T1D涉及多个项目，但在这笔赠款中，我们将 重点关注PTPN 22、TYK 2、SH2B3和IFIH 1四个编码变体。我们的工作和其他工作已经证明： TIDPTPN22风险变体（PTPN22R）改变抗原受体（AR）信号传导，影响T细胞选择， TYK2保护性变体（TYK2P）和SH2B3风险变体（SH2B3R）改变细胞因子 IFIH1风险变体（IFIH1R）导致IFN-1产生增强。在这款DP3中，我们将 解决PTPN22R、TYK2NP、SH2B3R变异体单独和组合有助于 T1D中致病性胰岛特异性T细胞的发展;这些变体的影响进一步 通过增强的干扰素反应放大，如IFIH1R所见，导致疾病风险增加 发展我们提出了以下具体目标：目标1将确定如何改变AR信号 与PTPN 22 R相关的程序导致胰岛特异性效应T细胞的致病性增加;目的2将 确定SH2B3R和TYK2P如何促进T细胞活化和分化程序的改变 目标3将确定PTPN22 R是否与信号程序协同作用 受SH2B3R、TYK2P或IFIH1R影响，以调节免疫耐受和T1D发展。我们将应用一个 综合方法利用：原代人类细胞，人类风险变体的鼠模型，并通过采取 最先进的基因编辑和蛋白质组学技术的优势。这种方法将使我们能够 全面询问这些特定基因的功能含义，以及信号传导如何影响这些基因的功能。 它们调节的程序协同作用以促成TID。值得注意的是，我们发现， 由T1D风险变体驱动的程序通常模仿在疾病中观察到的程序，即使在缺乏T1D风险变体的患者中也是如此。 我们对特定风险变量进行建模。因此，对特定遗传风险变异及其功能影响的研究 广泛适用于了解T1D的发病机制，并将为开发新的 用于治疗1型糖尿病的生物标志物和治疗剂。

ABSTRACT Type 1 diabetes (T1D) is a complex disease arising as a result of both genetic background and environmental exposures. Multiple biochemical and developmental programs have been implicated in its pathogenesis. While rare single gene defects can result in T1D, we hypothesize that T1D generally arises through a combination of synergistic defects in two or more key immune ‘programs’. Genetic variants associated with T1D can be used as a guide to understand how alterations in these programs individually and in combination contribute to the development and progression of T1D. Although multiple programs are implicated in T1D, in this grant we will focus on four coding variants, PTPN22, TYK2, SH2B3 and IFIH1. Our work and others has established that: the TID PTPN22 risk variant (PTPN22R) alters antigen receptor (AR) signaling, impacting T cell selection, development and survival; the TYK2 protective variant (TYK2P) and SH2B3 risk variant (SH2B3R) alter cytokine responses; and the IFIH1 risk variant (IFIH1R) results in enhanced IFN-1 production. In this DP3, we will address the hypothesis that PTPN22R, TYK2NP, SH2B3R variants individually and in combination contribute to the development of pathogenic islet specific T cells in T1D; and that the impact of these variants is further amplified by an enhanced Interferon response, as seen with IFIH1R, resulting in increased risk for disease development. We propose the following specific aims: Aim 1 will determine how the altered AR signaling program associated with PTPN22R leads to increased pathogenicity of islet specific effector T cells; Aim 2 will determine how SH2B3R and TYK2P contribute to alterations in the T cell activation and differentiation program in health and disease; and Aim 3 will determine whether PTPN22R synergizes with signaling programs impacted by SH2B3R, TYK2P or IFIH1R to modulate immune tolerance and T1D development. We will apply an integrated approach utilizing: primary human cells, murine models of the human risk variant, and by taking advantage of cutting edge gene editing and proteomics technologies. This approach will allow us to comprehensively interrogate the functional implications of these specific genes and, how the signaling programs that they modulate, function in concert to contribute to TID. Notably, we have found that the altered programs driven by T1D risk variants often mimic those observed in disease, even in patients lacking the specific risk variants we model. Thus, studies of specific genetic risk variants and their functional implications are broadly applicable to understanding the pathogenesis of T1D and will inform the development of new biomarkers and therapeutic agents to treat type 1 diabetes.