Using ex vivo, in vivo models and patient mutations to interrogate pancreatic exocrine-endocrine cross talk

项目概要 1 型糖尿病及其代谢后果仍然是最重要的生物医学问题之一 当今美国和全世界面临的挑战。除了与糖尿病特别相关的发病率外，该疾病 与肾衰竭、血脂紊乱、心血管疾病和癌症等并发症相关 全世界死亡的一个主要原因。因此迫切需要更好地了解发病机制 促进分泌胰岛素的β细胞的损失，以制定更好的疗法来对抗这种疾病。几个 研究认为，构成胰腺生态位的不同细胞类型有助于糖尿病的发病机制 疾病过程。从而更好地了解主要细胞之间的细胞间通讯，例如 因为腺泡、导管细胞和胰岛细胞是必要的。我们研究了突变型羧基酯的作用 脂肪酶（CEL）基因，在腺泡细胞中表达，其被β细胞吸收并导致其缺陷的能力 功能和生长。该提案的目标是发现腺泡分泌蛋白组时发生的变化 将导管细胞与人胰岛和β细胞一起孵育。这些研究的数据将产生新的 测试假设将允许更深入地研究人类腺泡、导管和 胰岛细胞。我们将在本提案中解决以下目标： 目标 1) 确定人类腺泡的能力 与人类导管细胞相比，直接影响胰岛细胞功能。我们将从人类中分离出电动汽车并对其进行表征 诱导多能干 (hIPS) 细胞衍生的腺泡与导管谱系定型类器官，并孵化 将它们与人类胰岛/β细胞和人类胰腺切片一起直接检查对胰岛细胞的影响 生物学。我们还将检查 EV 货物，特别关注源自转移 RNA 的片段。 目标 2) 我们将探讨由 hiPS 产生的腺泡衍生类器官衍生的 EV 的能力 来自MODY8患者的细胞，直接调节人类胰岛β细胞生物学。人类类器官的使用和 人类胰腺切片将为我们的研究提供转化相关性。得到的结果和数据集 这些实验将补充人类胰岛研究网络的努力，并提供新颖的 与更大的科学界共享资源。

PROJECT SUMMARY Type 1 diabetes and its metabolic consequences continue to be among the most significant biomedical challenges in the US and worldwide today. In addition to morbidities specifically related to diabetes the disease is associated with complications such as renal failure, lipid disorders, cardiovascular disease and cancer and is a major cause of death worldwide. Thus there is an urgent need for a better understanding of the pathogenesis that promotes the loss of insulin-secreting beta cells to plan better therapeutics to combat the disease. Several studies have argued that diverse cell types that make up the pancreatic niche contribute to the pathogenesis of the disease process. Thus a better understanding of the inter-cellular communication between major cells such as the acinar and duct cells and islet cells are warranted. We have studied the role of the mutant carboxy ester lipase (CEL) gene, expressed in acinar cells, on its ability to be taken up by beta cells and cause defects in its function and growth. The goal of this proposal is to discover the changes that occur when secretome from acinar versus duct cells are incubated with human islet and β-cells. The data from these studies will generate new hypothesis for testing that will allow a deeper interrogation of the cross-talk between human acinar, duct and islet cells. We will address the following Aims in this proposal: Aim 1) Determine the ability of human acinar versus human duct cells to directly impact islet cell function. We will isolate and characterize EVs, from human induced pluripotent stem (hIPS) cell derived acinar- versus duct-lineage committed organoids, and incubate them with human islet/β-cells and human pancreas slices to directly examine the consequences on islet cell biology. We will also examine the EV cargo with a particular focus on fragments derived from transfer RNAs. Aim 2) We will interrogate the ability of EVs, derived from acinar-derived organoids generated from the hiPS cells from MODY8 patients, to directly regulate human islet-β-cell biology. The use of human organoids and human pancreas slices will provide translational relevance of our studies. The results and datasets obtained from these experiments will complement the efforts of the Human Islet Research Network and provide novel resources to be shared with the larger scientific community.