Humanized Mouse Avatars for T1D

T1D 人性化鼠标头像

• 批准号: 10020970
• 负责人: Michael Allen Brehm
• 金额: $ 101.15万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-25 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10020970
• 关键词: Address; Animal Model; Antigens; Area; Autoantibodies; Autoimmune Process; Autologous; Beta Cell; Biological Assay; Biological Models; Cadaver; Cell Differentiation process; Cell Line; Cell Transplantation; Cells; Clinical Trials; Clone Cells; Cloning; Consent; Development; Diabetes Mellitus; Disease; Disease Progression; Engraftment; Ensure; Etiology; Genotype; Glucose; Goals; Grant; Hematopoietic stem cells; Human; Immune; Immune system; Immunodeficient Mouse; Immunotherapy; In Vitro; Individual; Insulin; Insulin-Dependent Diabetes Mellitus; Islets of Langerhans; Laboratories; MHC Class I Genes; Mediating; Methods; Modeling; Monitor; Mus; Natural History; Pathology; Patients; Phase; Plant Roots; Protocols documentation; Reagent; Reproducibility; Research Personnel; Resource Development; Rodent Model; Scientist; Source; Standardization; Stimulus; System; T-Lymphocyte; Therapeutic; Therapeutic Intervention; Thymic epithelial cell; Tissues; Transplantation; autoreactive T cell; autoreactivity; cell type; design; diabetic; human embryonic stem cell; human model; humanized mouse; in vivo; individual patient; induced pluripotent stem cell; interest; islet; meetings; multidisciplinary; novel; novel therapeutic intervention; novel therapeutics; patient subsets; prevent; reconstitution; recruit; risk variant; stem

PROJECT SUMMARY / ABSTRACT To date, studies of human type 1 diabetes (T1D) have failed to provide a mechanistic understanding of the underlying causes of the disease, largely because patients must be analyzed long after initiation of the autoimmune attack. Our ignorance of the key molecules and cells mediating the initiation and progression of human T1D may well underlie the paucity of significant new therapeutic interventions. This renewal proposal addresses the stated goal of RFA-DK-18-013 that “CMAI supports resource development projects that are primarily focused on the reagents and model systems needed for mechanistic study of human T1D.” Our Scientific Premise is that human T1D beta cells and immune cells transplanted into optimized immunodeficient mice (OPTI-MICE) will provide tractable model systems to study human T1D. We propose short term (Aim 1) and long term (Aim 2) goals. Aim 1 will develop an effector phase model of T1D in OPTI-MICE. Our team has already assembled the 3 key components that are required to develop an effector model of T1D: 1) Appropriate OPTI-MICE as recipients; 2) autoreactive T cells, lines, clones and iPS- derived T cells from T1D donors; 3) autologous human iPS cell-derived (SC)- cells. We have developed NSG mice deficient in MHC class I and II that do not develop GVHD when engrafted with functional human T cells for this aim. We have also recruited Dr. Sally Kent, an expert in islet autoreactive T cell cloning, to provide T1D autoreactive T cell lines and clones from consented T1D patients. Aim 2 will reconstruct human T1D in OPTI-MICE using cells derived from T1D iPS cells. These iPS cells will be used to produce the three key cell types: hematopoietic stem cells (HSC) that will generate immune systems, thymic epithelial cells (TEC), and β-cells, all integral to the pathology of T1D. These cells will be derived through the use of directed differentiation and reprogramming strategies. We have been successful in generating functional human β cells from human control and T1D patient iPS cells, providing a standardized and reproducible source of β cells for our studies. We have successfully performed directed differentiation of human ES cells to generate HSCs, and will use similar approachs for directed differentiation of iPS cells into HSCs. Functional human TEC will also be generated using directed differentiation protocols similar to those used to achieve fully differentiated human β cells. Each cell type will be subjected to rigorous analysis in vitro and in vivo to ensure full functionality. Differentiated β-cells, TECs, and immune cells derived from T1D donors will be co-transplanted into OPTI- MICE specifically optimized to enhance T cell, SC-β cell, SC-HSC, and SC-TEC cell engraftment and function allowing reconstitution of an individual patient's disease in an animal model. These new models of human diabetes will permit detailed observation, manipulation, and analysis of T1D, enabling us to determine the cells and antigens that initiate T1D, drive disease progression and mediate beta cell destruction. We have assembled an collaborative team of scientists that have the expertise required to accomplish this project.

项目摘要 /摘要 迄今为止，人类1型糖尿病（T1D）的研究未能提供对机械的理解 该疾病的根本原因，主要是因为必须在开始后很长时间分析患者 自动免疫攻击。我们对介导主动性和进展的关键分子和细胞的无知 人类T1D很可能是缺乏重大新治疗干预措施的基础。该更新建议 解决RFA-DK-18-013的既定目标，即CMAI支持资源开发项目 首先专注于人类T1D机械研究所需的试剂和模型系统。 科学的前提是人T1Dβ细胞和免疫细胞移植到优化的 免疫缺陷的小鼠（Opti-MICE）将提供可拖动的模型系统来研究人类T1D。我们 提案短期（目标1）和长期（目标2）目标。 AIM 1将开发一个效应子阶段模型 t1d在Opti-Mice中。我们的团队已经组装了开发一个需要的3个关键组件 T1D的效应器模型：1）适当的Opti-Mice作为接受者； 2）自动反应性T细胞，线，克隆和IPS- 从T1D供体派生的T细胞； 3）自体IPS细胞衍生（SC） - 细胞。我们已经开发了NSG 小鼠在与功能性人类T细胞植入的MHC I和II类中定义了GVHD 为此目标。我们还招募了胰岛自动反应性T细胞克隆专家Sally Kent博士，以提供 来自同意T1D患者的T1D自动反应性T细胞系和克隆。 AIM 2将重建人类T1D 使用源自T1D IPS细胞的细胞的Opti-erice。这些IPS单元将用于生成三个键 细胞类型：造血干细胞（HSC），将产生免疫系统，胸腺上皮细胞（TEC）， 和β细胞，都是T1D病理学的组成部分。这些单元将通过使用的使用来得出 分化和重编程策略。我们已经成功地产生了功能性人β细胞 从人类对照和T1D患者IPS细胞中，为β细胞的标准化和可重复的来源提供 我们的研究。我们已经成功地执行了人类ES细胞的定向分化，以产生HSC，并 将使用类似的方法将IPS细胞分化为HSC。功能性人类TEC也将是 使用与实现完全分化的人β相似的定向分化方案生成 细胞。每种细胞类型都将在体外和体内进行严格的分析，以确保完全功能。 从T1D供体衍生的分化的β细胞，TEC和免疫力将共同转移到光学中 专门优化以增强T细胞，SC-β细胞，SC-HSC和SC-TEC细胞植入和功能的小鼠 允许在动物模型中重建个体患者病。这些新的人类模型 糖尿病将允许对T1D的详细观察，操纵和分析，从而使我们能够确定细胞 以及启动T1D，驱动疾病进展并介导β细胞破坏的抗原。我们有 组建了一个由科学家组成的合作团队，他们具有完成该项目所需的专业知识。