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

使用离体、体内模型和患者突变来探究胰腺外分泌-内分泌串扰

• 批准号: 10706558
• 负责人: Saumya Das
• 金额: $ 91.95万
• 依托单位: JOSLIN DIABETES CENTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-20 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10706558
• 关键词: 2' -Deoxythymidine; Acinar Cell; Adult; Anatomy; Autophagocytosis; Beta Cell; Biological Models; Biology; Cardiovascular Diseases; Cause of Death; Cell Communication; Cell Line; Cell physiology; Cell secretion; Cells; Cellular Stress; Cellular biology; Cessation of life; Coculture Techniques; Communication; Communities; Complement; Data; Data Set; Defect; Development; Diabetes Mellitus; Disease; Disease Progression; Duct (organ) structure; Ductal Epithelial Cell; Embryonic Development; Endocrine; Esters; Exhibits; Exocrine pancreas; Extracellular Space; Functional disorder; Gene Mutation; Genes; Genetically Engineered Mouse; Goals; Growth; Hip region structure; Hormones; Human; Incubated; Insulin-Dependent Diabetes Mellitus; Islet Cell; Kidney Failure; Label; Lipase; Malignant Neoplasms; Malignant neoplasm of pancreas; Metabolic; Modeling; Molecular; Morbidity - disease rate; Mutation; Organoids; Outcome; Output; Pancreas; Pancreatic Diseases; Pancreatitis; Pathogenesis; Pathway interactions; Patients; Physiology; Process; Property; Reagent; Research; Resolution; Resources; Role; Secondary to; Slice; Small RNA; Testing; Therapeutic; Tissues; Transfer RNA; Transplantation; Variant; blood glucose regulation; cell type; combat; diabetes risk; endoplasmic reticulum stress; exosome; experimental study; extracellular vesicles; gene product; human disease; humanized mouse; in vitro Model; in vivo; in vivo Model; induced pluripotent stem cell; insight; insulin secretion; intercellular communication; islet; lipid disorder; mutant; non-diabetic; novel; paracrine; risk variant; senescence; stress granule; transmission process; uptake

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.

项目摘要 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。 在美国和世界范围内的挑战。除了与糖尿病特别相关的发病率外， 与并发症如肾衰竭、脂质紊乱、心血管疾病和癌症有关， 是全球死亡的主要原因。因此，迫切需要更好地了解其发病机制 促进胰岛素分泌β细胞的损失，以计划更好的治疗方法来对抗这种疾病。几 研究认为，构成胰腺小生境的不同细胞类型有助于胰腺炎的发病机制。 疾病的过程。因此，更好地理解主要细胞之间的细胞间通信， 因为腺泡和导管细胞以及胰岛细胞是必需的。我们研究了突变羧基酯的作用 脂肪酶（CEL）基因，在腺泡细胞中表达，对它被β细胞摄取的能力，并导致其缺陷， 功能和成长。这项提议的目的是发现当腺泡分泌蛋白质组在细胞内表达时， 与人胰岛和β细胞一起孵育。这些研究的数据将产生新的 假设的测试，这将允许更深入地询问人类腺泡，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和表征EV， 诱导多能干细胞（hIPS）来源的腺泡与导管谱系定向类器官，并孵育 它们与人胰岛/β细胞和人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，特别关注来自转移RNA的片段。 目的2）我们将询问EV的能力，EV来源于从hiPS产生的腺泡衍生的类器官 来自MODY 8患者的细胞，以直接调节人胰岛β细胞生物学。人类类器官的使用， 人胰腺切片将提供我们研究的翻译相关性。获得的结果和数据集 这些实验将补充人类胰岛研究网络的努力，并提供新的 与广大科学界共享资源。