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经开始阐明对建立β细胞的影响 胰岛的质量和形成，但遗传和环境的影响表现在出生后的胰腺 发展仍然是未知的。生命的第一个十年(本提案称为儿科阶段)是 胰腺发育中发生两个关键过程的动态时间：(1)β细胞团的建立和 (2)β细胞和胰岛功能成熟。此外，它还是第一次β细胞定向的1型自身免疫 糖尿病(T1D)通常开始。因此，了解支配儿科的分子和细胞过程 胰腺发育和功能是提高儿童青少年T1D诊断率的关键 和T2D，并开发预防或治疗β细胞功能障碍的策略。虽然正在进行的几项倡议 包括人类小岛研究网络(Hirn)一直在从成人非糖尿病患者中生成数据集， T1D和T2D供体，在胰腺深层分子和组织水平的表型方面存在重大差距 儿科时期。此外，血管、免疫和其他基质细胞群体的贡献 以及它们的β细胞相互作用对人类儿童胰腺发育的影响在很大程度上尚不清楚，尽管 它们对成人β细胞功能的已知影响。我们的建议是基于我们激动人心的单细胞多组体 空间分辨的试验数据，将使我们能够映射T1D的上下文专一性和相关的GWAs特性 胰腺中的信号跨越细胞类型、年龄、性别和发育阶段。此外，使用活体切片 技术，我们将能够用高密度原位研究细胞生理学和细胞间通讯。 时间分辨率，以深入了解管理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 同样的器官将提供前所未有的途径来定义分子特征、组织 结构，以及β细胞的成熟。这不仅是对从大多数成年人身上收集的表型数据的补充 人类胰腺分析计划(HPAP)中的捐赠者，但也将生成对几个HIRN有用的数据 财团和更广泛的研究社区。我们的多学科研究团队相互补充 精通胰腺和胰岛生物学、原位生理学、单细胞基因组学和表观基因组学、图像数据 分析、统计遗传学和机器学习设计了用于发现细胞状态动态的工具和分析 生命的第一个十年的变化，并定义这些变化如何影响下游生物 转录调控、胰腺内细胞空间组织和细胞功能。如果 这些研究的成功将为人类β的功能成熟提供新的机制见解 在关键的儿科生命阶段的细胞。这可能会影响我们对T1D发病机制的看法 并为糖尿病和其他胰腺疾病带来新的治疗方法。

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.