In situ analysis of functional endocrine, vascular, and immune cell interactions during early postnatal development of the human pancreas

如果1型糖尿病的病因尚不清楚，主要是因为人类的胰岛和它的 与免疫系统的相互作用还没有研究过。糖尿病研究界现在来了 承认在糖尿病啮齿动物模型中所获得的结果与人类状况缺乏相关性。在……里面 响应，有一种新的协调努力，以获取和研究相关材料，即人类 胰腺，在健康和疾病中。这项研究计划的长期目标是了解 以及人类胰岛在向糖尿病状态发展过程中发生的生理变化。这个 这项应用的目的是确定内分泌、血管和免疫部分是如何成熟的。 并在胰岛出生后的发育过程中进行功能上的相互作用。我们将重点关注青少年的成熟 这是因为在这一阶段，早发性自身免疫与 显性1型糖尿病的易感性。最重要的假设是，β细胞的启动是由 自身免疫与胰岛细胞分子表型的发育变化有因果关系 小岛建筑的变化。我们认为成熟过程使胰岛更容易发炎。 促进自身免疫的发展。提出这项研究的理由是理解 是什么让这个小岛变得脆弱，这不仅有助于解释它的衰落，还将为干预提供线索 战略。因此，该项目与美国国立卫生研究院的任务相关，并对研究目标作出反应。 NIDDK的资助机会公告题为“人类的高分辨率探索” 胰岛组织环境“。在初步数据的指导下，我们将通过追求三个具体的 目的：(1)确定胰岛内分泌细胞功能成熟的机制；(2)确定 建立血管功能的内分泌控制，并(3)确定表型和 胰岛驻留巨噬细胞的行为。在第一个目标下，我们将研究大规模的结构和功能 β细胞和阿尔法细胞充分发挥其分泌潜能所需的变化。在所有三个目标中，我们都将记录 功能成像的细胞反应和活体胰腺切片激素释放的测量 年龄从0岁到10岁。这些研究将得到scRNA-seq分析的补充 孤立的小岛。在第二个目标下，我们将确定内分泌细胞如何建立对 血管周细胞是胰岛血液流动的主要调节器。在第三个目标下，我们将研究如何 在胰岛成熟过程中，胰岛驻留巨噬细胞的表型和功能会发生变化。这个 拟议的研究具有重要意义，因为预期结果可能揭示出 削弱胰岛的自然防御，并引发当地免疫细胞的异常反应。了解这些 这一过程对于提出旨在预防1型糖尿病发展的干预目标至关重要。

If the causes of type 1 diabetes are not known it is mainly because the human pancreatic islet and its interactions with the immune system have not been studied. The diabetes research community is now coming to terms with the lack of relevance to the human situation of results obtained in rodent models of diabetes. In response, there is a new concerted effort at obtaining and studying the relevant material, namely the human pancreas, in health and disease. The long-term goal of this research program is to understand the anatomical and physiological changes that occur in the human islet during the progression towards the diabetic state. The objective of this application is to determine how the endocrine, vascular and immune compartments mature and interact functionally during the postnatal development of the islet. We will focus on the juvenile maturation period because it is a stage during which early-arising autoimmunity is strongly correlated with the predisposition towards overt type 1 diabetes. The overarching hypothesis is that the onset of beta cell-directed autoimmunity is causally related to developmental alterations in the molecular phenotypes of islet cells and to changes in islet architecture. We propose that maturation processes make islets susceptible to inflammation and facilitate the development of autoimmunity. The rationale for the proposed research is that understanding what makes the islet vulnerable will not only help explain its downfall but also provide clues for intervention strategies. This project is thus relevant to the mission of the NIH and is responsive to the research objectives of the Funding Opportunity Announcement from the NIDDK entitled “High-Resolution Exploration of the Human Islet Tissue Environment”. Guided by preliminary data, we will test our hypothesis by pursuing three specific aims: (1) determine the mechanisms of functional maturation of islet endocrine cells, (2) determine how endocrine control of vascular function is established, and (3) determine changes in the phenotype and behavior of islet resident macrophages. Under the first aim, we will study the massive structural and functional changes needed for beta and alpha cells to reach their full secretory potential. In all three aims, we will record cellular responses with functional imaging and measure hormone release in living pancreas slices from donors aged 0 to 10 years old. These studies will be complemented by scRNA-seq analyses of cells sorted from isolated islets. Under the second aim, we will determine how the endocrine cells establish control of the vascular pericyte, the major regulator of blood flow in the islet. Under the third aim, we will examine how the phenotype and function of the islet resident macrophages changes during the maturation of the islet. The proposed research is significant because the anticipated results could reveal developmental processes that diminish the islet’s natural defenses and trigger abnormal responses from local immune cells. Knowing these processes is crucial to propose intervention targets aimed at preventing the development of type 1 diabetes.