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Engineering islet-like organoids from gastric stem cells for T1D cell replacement therapy

从胃干细胞中改造胰岛样类器官，用于 T1D 细胞替代疗法

基本信息

批准号：
10512923
负责人：
Qiao Joe Zhou
金额：
$ 62.5万
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：
2022-09-12 至 2026-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10512923
关键词：
Adoption Anterior Antigens Attenuated Autoantigens Autoimmune Autoimmune Responses Autologous Autologous Transplantation Beta Cell Biological Assay Biological Response Modifiers Biopsy Biopsy Specimen CD47 gene CD8-Positive T-Lymphocytes Cell Communication Cell Line Cell Therapy Cells Cellular Assay Clone Cells Cytotoxic T-Lymphocytes Derivation procedure Diabetes Mellitus Doxycycline Engineering Future G6PC2 gene Generations Genetic Glucose Glucose tolerance test Human Human Cloning Immune Immune Evasion Immunosuppression In Vitro Innate Immune Response Insulin Insulin-Dependent Diabetes Mellitus Islets of Langerhans Transplantation Knock-in Laboratories Methods Molecular Mus NPM1 gene Names Organoids Persons Physiologic pulse Production Protocols documentation Stains Standardization Stomach Structure of beta Cell of islet T-Lymphocyte Technology Tissue Donors Tissue Grafts Tissues Transplantation Variant age group cell replacement therapy cell type clinical translation cytotoxic CD8 T cells diabetic eye chamber gastric organoids gastrointestinal glucose monitor glycemic control immunogenic improved in vivo in vivo imaging insulin secretion insulitis islet new technology preproinsulin progenitor programmed cell death ligand 1 single-cell RNA sequencing stem cells translational potential tumor

项目摘要

PROJECT SUMMARY Islet transplantation offers a potential cure for Type 1 diabetes (T1D). Wide adoption of a cell therapy requires abundant islet supplies and effective immune protection without long-term systemic immune suppression. My laboratory and others showed that it was feasible to derive insulin-secreting cells from renewable and abundant gastrointestinal (GI) stem cells. 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 samples and expanded them to billions. The hGSCs were engineered for transient activation of NGN3 and stable expression of PDX1 and MAFA (collectively referred to as NPM factors), leading to formation of thousands of GINS (Gastric Insulin Secreting) organoids. 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 with a scalable derivation method. GINS organoids contain 25-30% of cells that closely resemble pancreatic Beta-cells. In this project, we will evaluate the hypothesis that clonal hGSC lines yielding a higher percentage of Beta-like cells can be readily identified from donor tissues. We will develop a standard derivation protocol using genetic knockin of NPM and clonal selection with the aim to consistently produce highly functional GINS grafts from donors. GINS cells lack key autoantigens and may be naturally less immunogenic than islet Beta-cells. We will confer further autoimmune protection by constitutive expression of two potent immune regulators PD-L1 an CD47. Normal and immune-evasive organoids will be evaluated in vitro and in vivo with a panel of antigen-specific cytotoxic CD8 T cells, the main effector of Beta-cell demise in T1D. Together, these studies will create a technology for reliable production of autologous GINS grafts with strong autoimmune protection, suitable for long-term glycemic control without immune isolation or suppression.

项目总结