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的胰腺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起关键作用。 了解内在的和环境的网络签名动力学将有助于剖析分子 控制T1D进程的机制。在这里，我们建立了一支拥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进展过程中的环境变化。在初步研究中，我们进行了单细胞 转录组(scRNA-seq)和染色质(scATAC-seq)对健康、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 使用暴露在细胞因子或病毒下的人胰岛的细胞特异性特征。此外，我们还创建了一个 使用同基因人类多能干细胞(HPSCs)来源的胰腺β细胞和巨噬细胞的平台- 探索糖尿病相关细胞生物学功能相关基因或单核苷酸多态性。 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 系统研究关键的内在和环境信号动力学在T1D进展和 建立控制胰岛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： 目的1：确定T1D进展过程中细胞特异性的内在和环境特征。 目的2：破译控制T1D进展的细胞特异性基因调控网络。 目标3：验证T1D进展中的细胞、基因、遗传变异以及内在和环境特征 基于hPSC的同源平台和初级T1D胰岛。 我们的主要成果包括：1)单细胞分辨多组体(scRNA-seq，scATAC-seq)图谱， T1D和细胞因子或CVB4处理的人胰岛；2)细胞/上下文特异的分子遗传(e/caQTL) 人胰岛固有信号和环境信号的网络和中枢特征；3)同基因hPSC- 衍生的带有T1D相关(HUB)基因敲除的β细胞和免疫细胞，以验证单个或多个基因。 这些结果将为开发新药和精确的疾病进展标志物奠定基础。

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.