Determining the Intrinsic and Environmental Signal Contributing to Early T1D Progression

抽象的。 1 型糖尿病 (T1D) 是由自身免疫反应诱导胰腺 β 细胞破坏引起的。两者都是内在的 （β 细胞）和环境（免疫细胞）信号在胰腺 β 细胞功能障碍和死亡中发挥着关键作用。 了解内在和环境网络特征动态将有助于剖析分子 控制 T1D 进展的机制。在这里，我们建立了一支具有糖尿病专业知识的跨学科团队 计算和功能基因组学、干细胞生物学和胰岛生物学，系统地探索内在的 以及 T1D 进展期间的环境变化。在初步研究中，我们进行了单细胞 健康、自身抗体阳性（均为非 高血糖和高血糖）患者胰岛样本，以及模拟 T1D 胰岛的实验室模型 使用暴露于细胞因子或病毒的人类胰岛进行细胞特异性标记。此外，我们还创建了一个 使用同基因人类多能干细胞（hPSC）衍生的胰腺β细胞和巨噬细胞的平台- 像细胞一样探索糖尿病相关基因或单核苷酸多态性的生物学功能。 在这里，我们将结合我们在糖尿病计算和功能基因组学以及干细胞生物学方面的专业知识来 系统地研究关键内在和环境信号动力学在 T1D 进展中的作用， 建立控制胰腺β细胞功能障碍的机制网络。为了实现这些目标，我们 提出三个具体目标： 目标 1：确定 T1D 进展过程中细胞特异性的内在和环境特征。 目标 2：解码控制 T1D 进展的细胞特异性基因调控网络。 目标 3：使用 T1D 进展验证细胞、基因、遗传变异以及内在和环境特征 基于同基因 hPSC 的平台和原代 T1D 胰岛。 我们的主要成果包括：1) 健康、健康的单细胞分辨率多组学（scRNA-seq、scATAC-seq）图谱 T1D 和细胞因子或 CVB4 处理的人胰岛； 2) 细胞/环境特异性分子遗传学 (e/caQTL) 人类胰岛固有信号和环境信号的网络和集线器特征； 3) 同基因 hPSC- 衍生的 β 细胞和免疫细胞具有 T1D 相关（中心）基因敲除，以验证单个或多个基因。 这些结果将为开发新药和精准疾病进展标记物奠定基础。

Abstract. Type 1 diabetes (T1D) is caused by autoimmune response induced pancreatic β cell destruction. Both intrinsic (beta cell) and environmental (immune cell) signals play critical roles in pancreatic β cell dysfunction and death. Understanding the intrinsic and environmental network signature dynamics will facilitate dissecting the molecular mechanisms controlling T1D progression. Here, we build an interdisciplinary team with the expertise of diabetes computational and functional genomics, stem cell biology, and islet biology to systematically explore the intrinsic and environmental changes during T1D progression. In the preliminary studies, we performed single cell transcriptome (scRNA-seq) and chromatin (scATAC-seq) profiling of healthy, autoantibody positive (both non- hyperglycemia and hyperglycemia) patient islet samples, as well as a laboratory model that mimics T1D islet cell-specific signatures using human islets exposed to either cytokines or virus. In addition, we have created a platform to use isogenic human pluripotent stem cells (hPSCs)-derived pancreatic beta cells and macrophage- like cells to explore the biological function of diabetes associated genes or single nucleotide polymorphisms. Here, we will combine our expertise of diabetes computational and functional genomics and stem cell biology to systematically investigate the role of key intrinsic and environmental signal dynamics in T1D progression and establish the mechanistic network controlling pancreatic beta cell dysfunction. To achieve these goals, we propose three specific aims: Aim 1: Determine the cell-specific intrinsic and environmental signatures during T1D progression. Aim 2: Decode the cell-specific genetic regulatory network controlling T1D progression. Aim 3: Validate cells, genes, genetic variants, and intrinsic and environment signatures in T1D progression using an isogenic hPSC-based platform and primary T1D islets. Our key deliverables include: 1) a single-cell resolution multi-omic (scRNA-seq, scATAC-seq) map of healthy, T1D and cytokine- or CVB4 treated human islets; 2) the cell/context-specific molecular genetic (e/caQTL) network and hub signature of intrinsic and environmental signals in human pancreatic islets; 3) Isogenic hPSC- derived beta and immune cells with T1D-associated (hub) gene knockouts to validate individual or multiple genes. These results will be foundational for development of novel drugs and precision disease progression markers.