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

摘要 人类胰腺发育的研究已经开始阐明β细胞建立的影响 胰岛的质量和形成，但出生后胰腺期间表现出遗传和环境的影响 发展仍然未知。生命的第一个十年（本建议称为儿科期）是一个 胰腺发育的动态时间，发生两个关键过程：（1）β细胞团的建立， (2)β细胞和胰岛功能成熟。此外，这是时间β细胞导向的自身免疫1型 糖尿病（T1 D）通常会出现。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T1 D诊断的关键 和2型糖尿病以及发展预防或治疗β细胞功能障碍的策略。虽然一些正在进行的倡议 包括人类胰岛研究网络（HIRN）已经从成人非糖尿病患者中生成数据集， T1 D和T2 D供体，在胰腺的深层分子和组织水平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的贡献， 和它们的β细胞相互作用，对人类儿科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。我们的建议是基于我们令人兴奋的单细胞多组学 空间分辨的试点数据，使我们能够映射T1 D和相关性状GWAS的背景特异性 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，我们将能够研究细胞生理学和细胞间通讯原位与高 时间分辨率，以提供对控制β细胞成熟和 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 结构和β细胞成熟。这不仅将补充从大多数成年人中收集的表型数据， 人类胰腺分析计划（HPAP）中的供体，但也将产生对几个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与互补 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，以发现细胞状态动态 在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学， 转录调控、胰腺内的细胞空间组织和细胞功能。如果 这些研究将为人类β-淀粉样蛋白的功能成熟提供新的机制见解。 在关键的儿科生命阶段。这可能会影响我们对T1 D发病机制的认识 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.