Integrative analysis of multi-omic signatures and cellular function in human pancreas across developmental timeline at single-cell spatial resolution

摘要 对人类胰腺发育的研究已开始阐明对β细胞建立的影响 胰岛的质量和形成，但出生后胰腺中表现出的遗传和环境影响 发展仍是未知数。生命的第一个十年（本提案中称为儿科时期）是 当两个关键过程发生时，胰腺发育的动态时间：(1) β细胞团建立并且 (2) β细胞和胰岛功能成熟。另外，现在是β细胞定向的1型自身免疫的时间 糖尿病（T1D）通常会开始。因此，了解控制儿科的分子和细胞过程 胰腺发育和功能是改善儿童和青少年 T1D 诊断的关键 和 T2D 并制定预防或治疗 β 细胞功能障碍的策略。尽管有多项正在进行的举措 包括人类胰岛研究网络 (HIRN) 一直在生成成人非糖尿病数据集， T1D 和 T2D 供体中，胰腺的深层分子和组织水平表型分析存在重大差距 小儿时期。此外，血管、免疫和其他基质细胞群的贡献 及其 β 细胞相互作用对人类小儿胰腺发育的影响在很大程度上是未知的，尽管 它们对成人 β 细胞功能的已知影响。我们的提案基于我们令人兴奋的单细胞多组学 空间解析的试点数据将使我们能够绘制 T1D 的背景特异性和相关特征 GWAS 胰腺中的信号跨越细胞类型、年龄、性别和发育阶段。此外，使用活切片 技术，我们将能够以高通量原位研究细胞生理学和细胞间通讯 时间分辨率可深入了解控制 β 细胞成熟和建立的过程 健康的胰腺结构。空间、生理、转录和染色质数据的叠加 相同的器官将提供前所未有的途径来定义分子特征、组织的变化 结构和 β 细胞成熟。这不仅可以补充从大多数成人中收集的表型数据 人类胰腺分析计划 (HPAP) 的捐赠者，但也将生成对多个 HIRN 有用的数据 联盟和更广泛的研究界。我们的多学科研究团队具有互补性 胰腺和胰岛生物学、原位生理学、单细胞基因组学和表观基因组学、图像数据方面的专业知识 分析、统计遗传学和机器学习设计的工具和分析来发现细胞状态动态 生命第一个十年的变化，并定义这些变化如何影响下游生物学 转录调节，对胰腺内的细胞空间组织和细胞功能。如果 如果成功，这些研究将为人类β功能成熟提供新的机制见解 儿童关键生命阶段的细胞。这可能会影响我们对 T1D 发病机制的看法 并导致糖尿病和其他胰腺疾病的新疗法。

Abstract Studies of human pancreas development have begun to elucidate influences in the establishment of β cell mass and formation of islets, but genetic and environmental influences that manifest during postnatal pancreas development remain unknown. The first decade of life (termed the pediatric period for this proposal) is a dynamic time in pancreas development when two critcal processes occur: (1) β cell mass is established and (2) β cells and islets functionally mature. In addition, it is the time β cell-directed autoimmunity of type 1 diabetes (T1D) often begins. Thus, understanding the molecular and cellular processes that govern pediatric pancreas development and function is key to improving the diagnosis of children and adolescents with T1D and T2D and developing strategies to prevent, or treat the β cell dysfunction. While several ongoing initiatives including the Human Islet Research Network (HIRN) have been generating datasets from adult nondiabetic, T1D, and T2D donors, there is a major gap in deep molecular and tissue-level phenotyping of pancreata from the pediatric period. Furthermore, the contributions of vascular, immune, and other stromal cell populations and their β cell interactions, to human pediatric pancreas development are largely uncharacterized, despite their known influence on adult β cell function. Our proposal is based on our exciting single-cell multi-omic spatially-resolved pilot data that will allow us to map the context specificity of T1D and related trait GWAS signals in pancreas across cell type, age, sex, and developmental stage. Moreover, using living slice technology, we will be able to investigate cellular physiology and cell-cell communication in situ with high temporal resolution to provide an insight into processes that govern β cell maturation and establishment of healthy pancreatic architecture. The overlay of spatial, physiological, transcriptional, and chromatin data from the same organs will provide unprecedented access to define changes in molecular signatures, tissue architecture, and β cell maturation. This will not only complement phenotypic data collected from mostly adult donors in the Human Pancreas Analysis Program (HPAP), but will also generate data useful to several HIRN consortia and the broader research community. Our multidisciplinary research team with complementary expertise in pancreas and islet biology, in situ physiology, single cell genomics and epigenomics, image data analysis, statistical genetics, and machine learning devised tools and analyses to discover cell state dynamic changes across the first decade of life and define how these changes influence downstream biology from transcriptional regulation, to cellular spatial organization within the pancreas, and cellular function. If successful, these studies will provide new mechanistic insights about the functional maturation of human β cells during the critical pediatric life stages. This will likely influence the way we perceive T1D pathogenesis and lead to new therapies for diabetes and other pancreas diseases.