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 胰岛素分泌β细胞移植是1型糖尿病（T1 D）的有效治疗方法。到 为了确保这些治疗细胞的供应，许多研究小组正在积极优化方案， 通过人胚胎干细胞（hESC）的分化或通过 重组成人组织下一个主要挑战是找到保护 从自身免疫移植细胞，以实现长期血糖控制。临床观察结果 长期以来一直认为T1 D患者中存在逃避自身免疫的β细胞， 继续发挥作用。从临床样品中研究这些细胞非常具有挑战性。新的研究 需要工具来解决如何产生胰岛素分泌细胞的关键问题， 抵抗自身免疫。我们的实验室最近发现，胰岛素分泌细胞可以很容易地 使用直接重编程方法从鼠胃组织产生， 因子Ngn 3、Pdx 1和Mafa（称为NPM因子）。胃源性胰岛素+细胞 胰腺β细胞的关键分子和功能特征，但与天然β细胞不同 细胞我们将胃胰岛素+细胞移植到T1 D的NOD小鼠模型中， 令人惊讶的观察是，胃胰岛素+细胞，不像天然的胰腺β细胞，不受 强烈的免疫攻击，表明免疫原性降低。这一观察提出了 令人兴奋的可能性，胃源性胰岛素+细胞可以作为一种新的工具，研究 免疫-β细胞相互作用在人类T1 D，并可进一步发展成移植 需要最低限度免疫保护的疗法。在本提案中，我们将开发新方法， 从人胃组织中产生功能性胰岛素分泌细胞， 方法和研究其免疫原性的各种工具，包括人类T1 D- 衍生的T细胞克隆。我们的最终目标是获得对免疫反应的机制性见解， T1 D中的β细胞，并发现可能减少或预防自身免疫性疾病的新途径。 攻击如果成功，该项目还将为一种新的胰岛素分泌来源奠定基础。 细胞用于治疗性移植以治疗T1 D。