Dissecting transcriptomics and epigenomic signatures of immune cells in type 1 diabetes

T1D是一种自身免疫性疾病，其中细胞毒性T细胞攻击并破坏分泌胰岛素的胰腺β 细胞遗传学在T1D发展中的作用从其在家庭中的聚集是显而易见的。虽然全基因组 关联研究揭示了T1D相关的单核苷酸多态性，目前有大量的 关于遗传学促进自身免疫的分子过程的知识存在差距。 具体来说，由于大多数疾病相关的SNP都存在于非编码基因组区域， 被认为是影响基因调控，而不是导致突变蛋白质的产生。的最新进展 我们对核组织的理解表明，遗传变异可能通过以下方式影响基因调控： 改变的3D基因组结构和重组的大转录协调区域的基因组， 疾病相关细胞类型。然而，序列变异、细胞环境、3D基因组之间的联系 T1D中的异常组织和异常基因表达在很大程度上仍然未知。我们的总体目标是定义 人胰腺组织中T1D相关免疫细胞的分子标志，并研究 这种深度剖析在检测早期T1D过程。我们的最新结果首次提供了早期 转录组学和T1D中的3D表观基因组改变。我们的假设是致病性免疫细胞亚型 存在于T1D的无症状和临床诊断阶段的胰腺组织中， 和3D表观基因组签名。我们建议产生最深入的可能的免疫分子谱 临床上充分表征的人类器官供体中胰腺组织和外周血中的细胞群 由HPAP收集。由于生物标志物检测最容易获得的实体，即血液，是 主要的免疫运输发生时，我们将检查β细胞破坏的免疫学特征是否可以 在T1D的无症状阶段的外周血中发现。一旦分子和表观基因组特征 T1D在胰腺组织中被精确定义，它们可以用作强大的磁铁来寻找针头 在循环细胞的干草堆中。我们的单细胞分辨率实验将识别T1D相关免疫 胰腺组织中的细胞和失调基因（Aim 1）。我们最先进的表观基因组学成像技术 基因组工程技术将确定与T1D相关的3D基因组错误折叠事件（Aim 2）。 每个目标的结果可以大大扩展我们对早期疾病过程的理解。的 整合这两个目的所获得的知识可以阐明致病基因的详细分子机制 在T1D中。

T1D is an autoimmune disease in which cytotoxic T cells attack and destroy insulin-secreting pancreatic beta cells. The role of genetics in T1D development is evident from its clustering in families. Although genome-wide association studies uncovered the T1D-associated single-nucleotide-polymorphisms, currently there is a large gap in knowledge regarding the molecular processes through which genetics contributes to autoimmunity. Specifically, because most disease-associated SNPs have been found in non-coding genomic regions, they are thought to impact gene regulation rather than causing production of mutated proteins. Recent advances in our understanding of nuclear organization indicate that genetic variation may impact gene regulation through altered 3D genomic structure and reorganization of large transcriptionally coordinated regions of the genome in the disease relevant cell type(s). However, the link between sequence variation, cellular context, 3D genome organization, and aberrant gene expression in T1D remains largely unknown. Our overall objective is to define the molecular hallmark of T1D-associated immune cells from human pancreatic tissues and study the utility of such deep profiling in detecting early T1D processes. Our latest results provide the first-ever evidence of early transcriptomics and 3D epigenomic alterations in T1D. Our hypothesis is that pathogenic immune cell subtypes residing in pancreatic tissues in asymptomatic and clinically diagnosed phases of T1D share transcriptional and 3D epigenomic signatures. We propose to generate the deepest-possible molecular profiling of immune cell populations in pancreatic tissues and peripheral blood in clinically well-characterized human organ donors collected by HPAP. Since the most accessible entity for biomarker testing, i.e. blood, is the conduit by which major immunological traffic occurs, we will examine if immunological features of beta cell destruction can be found in peripheral blood at asymptomatic stages of T1D. Once the molecule and epigenomic signatures of T1D are precisely defined in pancreatic tissues, they can be used as a powerful magnet to look for the needle in the haystack of circulating cells. Our single-cell resolution experiments will identify T1D-associated immune cells and deregulated genes in pancreatic tissues (Aim 1). Our state-of-the-art epigenomics, imaging, and genome engineering techniques will determine 3D genome misfolding events associated with T1D (Aim 2). The outcomes of each Aim can dramatically expand our understanding of early disease processes. The integration of knowledge gained in two Aims can elucidate detailed molecular mechanisms of pathogenic gene regulation in T1D.