Derivation of pancreatic islet-like organoids from human gastric stem cells

从人胃干细胞中衍生胰岛样类器官

基本信息

批准号：
10502451
负责人：
Qiao Joe Zhou
金额：
$ 60.21万
依托单位：
WEILL MEDICAL COLL OF CORNELL UNIV
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-08-24 至 2026-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10502451
关键词：
ATAC-seq Adoption Adult Autopsy Beta Cell Binding Biopsy Blood Glucose Cell Line Cells Chromatin Clone Cells D Cells Data Derivation procedure Diabetes Mellitus Diabetic mouse Enhancers Frequencies Functional disorder G Cells GCG gene Genes Genetic Transcription Genomics Glucagon Glucose Glucose tolerance test Goals Human Human Cloning Immune In Vitro Insulin Insulin-Dependent Diabetes Mellitus Islet Cell Islets of Langerhans Islets of Langerhans Transplantation Knock-out Laboratories Measures Messenger RNA Methods Mus NPM1 gene Names Organoids Pancreas Patients Production Sampling Stains Stomach Technology Tissue Grafts Tissues Transcriptional Activation Transplantation base blood glucose regulation cell type clinical translation diabetic gastrointestinal glucose monitor improved improved functioning in vivo insight insulin secretion islet nanoparticle new technology novel paracrine reconstitution response single-cell RNA sequencing stem cells tool translational potential tumor

项目摘要

PROJECT SUMMARY Islet transplantation offers a potential cure for Type 1 diabetes (T1D). Wide adoption of this promising therapy requires abundant islet supplies and effective immune protection. My laboratory and others showed that it was feasible to derive insulin-secreting cells from gastrointestinal (GI) tissues. However, it has not been possible to mass-produce islet-like organoids from human GI tissues for detailed assessment of their translational potential. In preliminary studies, we established methods to culture human gastric stem cells (hGSCs) from biopsy or autopsy samples that can be expanded to billions. We developed a scalable 2-step method to produce thousands of GINS (Gastric Insulin Secreting) organoids by transient activation of NGN3 and stable expression of PDX1 and MAFA (collectively referred to as NPM factors). GINS organoids acquired glucose-stimulated-insulin-secretion (GSIS) within 10 days, and upon transplantation, rapidly reversed diabetes in mice and maintained normoglycemia for over 3 months, with no tumor formation. Human GINS organoids thus have favorable attributes as a potential cell product for T1D treatment. GINS organoids contain 25% of cells that closely resemble pancreatic E-cells but a paucity of GCG+ and SST+ cells. Human islets have 50-75% E-cells, 25-35% D-cells, and 5% G-cells. Both D- and G- cells exert paracrine effects on E-cell section. In this project, we aim to develop new clonal hGSC lines and novel nanoparticle-based mRNA transduction method suitable for mass production of organoids that closely mimic human islets in cell composition and function. These studies are based on preliminary data indicating that hGSC clonal lines are markedly different, with some predominantly producing E-like or D-/G-like cells. Their differentiated progenies can thus be combined to yield islet- like organoids. We will further study the clonal lines for chromatin features and PDX1/MAFA genomic binding to gain mechanistic insight in GINS formation. Together, these studies constitute a major step in advancing the long-term goal of developing GINS organoids for T1D treatment.

项目概要胰岛移植为 1 型糖尿病 (T1D) 提供了潜在的治愈方法。这种有希望的广泛采用治疗需要充足的胰岛供应和有效的免疫保护。我的实验室和其他人表明从胃肠道（GI）组织中提取胰岛素分泌细胞是可行的。然而，目前还不可能从人类胃肠道组织中大规模生产胰岛样类器官。详细评估其转化潜力。在初步研究中，我们建立了培养人胃干细胞（hGSC）的方法活检或尸检样本可扩大至数十亿。我们开发了一种可扩展的两步方法通过 NGN3 的瞬时激活产生数千个 GINS（胃胰岛素分泌）类器官 PDX1和MAFA（统称为NPM因子）的稳定表达。杜松子酒类器官在 10 天内获得葡萄糖刺激胰岛素分泌 (GSIS)，并且在移植后，快速逆转小鼠糖尿病并维持正常血糖超过3个月，且无肿瘤形成。因此，人类 GINS 类器官具有作为 T1D 潜在细胞产品的有利属性治疗。 GINS 类器官含有 25% 的细胞与胰腺 E 细胞非常相似，但缺乏 GCG+ 和 SST+ 细胞。人类胰岛含有 50-75% E 细胞、25-35% D 细胞和 5% G 细胞。 D-和G- 细胞对 E 细胞切片发挥旁分泌作用。在这个项目中，我们的目标是开发新的克隆 hGSC 线和新型基于纳米颗粒的 mRNA 转导方法，适合大规模生产在细胞组成和功能上非常模仿人类胰岛的类器官。这些研究基于初步数据表明 hGSC 克隆系明显不同，其中一些主要产生E样或D-/G样细胞。因此，它们的分化后代可以组合产生胰岛- 像类器官。我们将进一步研究克隆系的染色质特征和 PDX1/MAFA 基因组结合以获得杜松子酒形成的机制见解。这些研究共同构成了重要的一步推进开发用于 T1D 治疗的 GINS 类器官的长期目标。