Generating novel sources of functional human insulin-secreting cells for T1D modeling

为 T1D 建模生成功能性人胰岛素分泌细胞的新来源

• 批准号: 9459621
• 负责人: Qiao Joe Zhou
• 金额: $ 72.22万
• 依托单位: HARVARD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-20 至 2018-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9459621
• 关键词: Address; Adult; Animals; Antral; Autoimmune Diabetes; Autoimmune Process; Autoimmunity; Beta Cell; Biopsy Specimen; Blood Glucose; Cell Communication; Cell Differentiation process; Cell Survival; Cell Transplants; Cells; Chemicals; Clinical; Clone Cells; Complement; Diabetes Mellitus; Enteroendocrine Cell; Epithelial; Foundations; Gastric Tissue; Genes; Genetic; Genetic Engineering; Glucose; Goals; Harvest; Hormone use; Human; Hyperglycemia; Immune; Immune response; Immunohistochemistry; In Vitro; Inbred NOD Mice; Insulin; Insulin-Dependent Diabetes Mellitus; Laboratories; Measures; Mesenchymal; Methods; Modeling; Molecular; Monitor; Mus; NPM1 gene; Natural Killer Cells; Organ; Pathway interactions; Patients; Production; Property; Protocols documentation; Research; Research Personnel; SCID Mice; Sampling; Secure; Signal Pathway; Source; Stem cells; Stomach; Structure of beta Cell of islet; T-Lymphocyte; Technology; Testing; Therapeutic; Thyroid Hormones; Time; Tissues; Transplantation; Tretinoin; Work; adeno-associated viral vector; base; diabetic; effective therapy; embryonic stem cell; glycemic control; human embryonic stem cell; immunogenic; immunogenicity; insight; insulin secretion; macrophage; mouse model; novel; prevent; three dimensional cell culture; tool

PROJECT SUMMARY Transplantation of insulin-secreting beta cells is an effective therapy for Type 1 diabetes (T1D). To secure supplies of these therapeutic cells, many research groups are actively optimizing protocols to derive insulin+ cells by differentiation of human embryonic stem cells (hESCs) or by reprogramming of adult tissues. The next major challenge is to discover methods to protect transplanted cells from autoimmunity to achieve long-term glycemic control. Clinical observations have long suggested existence of beta cells in T1D patients that escape autoimmunity and continue to function. Such cells are very challenging to study from clinical samples. New research tools are needed to address the critical question of how to produce insulin-secreting cells that resist autoimmunity. Our laboratory recently discovered that insulin-secreting cells can be readily produced from murine gastric tissues using a direct reprogramming approach with defined genetic factors Ngn3, Pdx1, and Mafa (referred to as NPM factors). The gastric-derived insulin+ cells share key molecular and functional features of pancreatic beta cells but are not identical to native beta cells. We transplanted gastric insulin+ cells into the NOD murine model of T1D and made the surprising observation that gastric insulin+ cells, unlike native pancreatic beta cells, are not subject to strong immune attack, suggesting reduced immunogenic properties. This observation raised the exciting possibility that gastric-derived insulin+ cells could be employed as a novel tool to study immune-beta cell interactions in human T1D and may be further developed into a transplantation therapy that requires minimal immune protection. In this proposal, we will develop new methods to produce functional insulin-secreting cells from human gastric tissues using two separate approaches and study their immunogenic properties with a variety of tools including human T1D- derived T-cell clones. Our ultimate goal is to gain mechanistic insight into the immune response to beta cells in T1D and to discover novel pathways that may diminish or prevent the autoimmune attack. If successful, this project will also lay the foundation for a novel source of insulin-secreting cells for therapeutic transplantation to treat T1D.

项目摘要 分泌胰岛素β细胞的移植是1型糖尿病（T1D）的有效疗法。到 这些治疗细胞的安全供应，许多研究小组正在积极优化方案 通过分化人类胚胎干细胞（HESC）或通过 重编程成人组织。下一个主要挑战是发现保护方法 从自身免疫的移植细胞获得长期血糖控制。临床观察 长期以来，在T1D患者中存在β细胞的存在，这些患者逃脱了自身免疫性和 继续运行。从临床样品中研究此类细胞非常具有挑战性。新研究 需要工具来解决如何产生分泌胰岛素分泌细胞的关键问题 抵抗自身免疫性。我们的实验室最近发现，分泌胰岛素的细胞很容易 使用直接重编程方法从鼠胃组织产生 因子NGN3，PDX1和MAFA（称为NPM因素）。胃来源的胰岛素+细胞共享 胰腺β细胞的关键分子和功能特征，但与天然beta并不相同 细胞。我们将胃胰岛素+细胞移植到T1D的点头鼠模型中，并使 令人惊讶的是，与天然胰腺β细胞不同的是，胃胰岛素+细胞不是对 强烈的免疫发作，表明免疫原性降低。这种观察提出了 胃衍生的胰岛素+细胞可以用作研究的新工具的令人兴奋的可能性 人类T1D中的免疫β细胞相互作用，可以进一步发展为移植 需要最小的免疫保护的治疗。在此建议中，我们将开发新方法 使用两个独立的 使用多种工具，包括人类T1D- 派生的T细胞克隆。我们的最终目标是获得对免疫反应的机械洞察力 T1D中的β细胞并发现可能减少或预防自身免疫的新型途径 攻击。如果成功，该项目还将为胰岛素分泌的新来源奠定基础 用于治疗T1D的治疗移植细胞。