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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：
10689788
负责人：
Qiao Joe Zhou
金额：
$ 58.34万
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/10689788
关键词：
Adoption Adult Alpha Cell 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 GCG gene Genes Genetic Transcription Genomics Glucagon Glucose Glucose tolerance test Goals Human Immune In Vitro Insulin Insulin-Dependent Diabetes Mellitus Islets of Langerhans Islets of Langerhans Transplantation Knock-out Laboratories Maps Measures Messenger RNA Methods Mus NPM1 gene Names Organoids Patients Production Proliferating Sampling Stains Stomach Structure of beta Cell of islet Technology Tissue Grafts Tissues Transcriptional Activation Transplantation blood glucose regulation cell type clinical translation diabetic gastrointestinal glucose monitor improved in vivo insight insulin secretion islet large scale production 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 β-cells but a paucity of GCG+ and SST+ cells. Human islets have 50-75% β-cells, 25-35% α-cells, and 5% δ-cells. Both α- and δ- cells exert paracrine effects on β-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 β-like or α-/δ-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.

项目总结