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

T1D是一种自身免疫性疾病，细胞毒性T细胞攻击并破坏分泌胰岛素的胰岛β 细胞。遗传学在T1D发育中的作用从其在家族中的聚集可见一斑。尽管全基因组 协会研究发现了与T1D相关的单核苷酸多态，目前有很大的 关于遗传学促进自身免疫的分子过程的知识差距。 具体地说，因为大多数与疾病相关的SNPs都是在非编码基因组区域发现的，所以它们 被认为影响基因调控，而不是导致突变蛋白的产生。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途径影响基因调控 大鼠基因组3D结构的改变和大转录协调区的重组 疾病相关细胞类型(S)。然而，序列变异、细胞背景、3D基因组之间的联系 T1D中的异常基因表达在很大程度上仍不清楚。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及其应用研究 在检测早期T1D过程中如此深入的剖析。我们的最新研究结果首次证明了 T1D的转录和3D表观基因组改变。我们的假设是致病免疫细胞亚型 在无症状和临床诊断的T1D共享转录阶段驻留在胰腺组织中 和3D表观基因组签名。我们建议产生免疫的最深可能的分子图谱 临床特征良好的人体器官供者胰腺组织和外周血中的细胞群 由HPAP收集。由于生物标记物检测最容易获得的实体，即血液，是通过其 发生重大免疫流量时，我们将检查是否有免疫学特征的β细胞被破坏 在T1D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精确地定义在胰腺组织中，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里。我们的单细胞分辨实验将识别T1D相关免疫 胰腺组织中的细胞和非调控基因(目标1)。我们最先进的表观基因组学、成像和 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)。 每个目标的结果都可以极大地扩展我们对早期疾病过程的理解。这个 整合这两个目的的知识可以阐明致病基因的详细分子机制 T1D中的法规。

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.