Immunogenicity of Human Stem Cell-Derived Beta Cells and Muscle Cells in Humanized Mice

人胚胎干细胞（ESC）和人诱导多能干细胞（iPSC）的开发 释放了细胞疗法治疗多种人类疾病的潜力。替代疗法 使用人干细胞衍生的β（SC-β）和SC-成肌细胞（本项目中使用的两种细胞类型），以及 基本上NIH的所有研究所和中心都对多种其它干细胞衍生物感兴趣。到 避免排斥，细胞替代疗法将需要通过包封的免疫保护， 免疫逃避特性，或与免疫抑制一起给予。由于细胞产物是人源性的， 由于有效异种移植，免疫原性不能使用免疫活性动物模型在体内测试 应答人们可以测试细胞的安全性，有效性和免疫原性，这将是时间 消耗、昂贵且具有潜在危险。因此，翻译人类语言的一个关键要求是， 以干细胞为基础的治疗走向临床是发展稳健的临床前动物模型进行评价 有效性，安全性，重要的是，人类细胞疗法的免疫原性。我们建议修改 人糖尿病和肌肉疾病的NOD-scid IL 2 rgnull（NSG）模型，以允许开发稳健的 人体免疫系统这将允许快速临床前评价的功能和免疫原性， 人SC-β细胞和SC-成肌细胞（以及正在开发的其他模型中的SC衍生细胞）。我们有 证明了目前移植人类免疫系统的NSG小鼠模型不能完全排斥 同种异体人ESC或SC-内皮细胞，并且在初步研究中，不能排斥人SC-β细胞。 我们的科学假设是，移植的人类免疫系统的功能有限， 人源化NSG小鼠的免疫缺陷是由于次级淋巴组织中缺乏有组织的结构。我们有 观察到新生NSG小鼠与野生型鼠HSC的移植将导致 淋巴结和强大的免疫反应，这表明缺乏物种特异性因素阻碍了 人源化NSG小鼠的淋巴发育。在这个多PI，多学科团队提案中，目标1将 产生表达人类特异性因子的糖尿病和肌肉疾病的新型NSG小鼠品系 支持淋巴结构和人体免疫功能。目的2将人SC-β细胞和SC-β细胞移植到体外， 我们独特的糖尿病和肌肉疾病的人源化小鼠模型， 和免疫原性。目标3将向杰克逊实验室（JAX）提供这些经验证的新菌株 用于全球分销的生物储存库。我们的建议利用强大的新技术， 创建新的人源化小鼠模型，并建立在我们的生成，验证和 迄今为止，自2005年以来，通过JAX分销网络共享了17种新的人源化NSG小鼠模型。 我们相信，我们的创新方法，结合我们的多学科合作团队将 确保这些急需的再生医学临床前模型的发展。

Development of human embryonic stem cells (ESCs) and human induced pluripotent stem cells (iPSCs) has unleashed the potential of cellular therapy for a number of human diseases. Replacement therapies using human stem cell-derived β (SC-β) and SC-myoblasts (the two cell types used in this project), and multiple other stem cell derivatives are of interest to essentially all of the Institutes and Centers at NIH. To avoid rejection, cell replacement therapies will require immune protection by encapsulation, provided immune-evading properties, or given with immunosuppression. As the cell products are of human origin, immunogenicity can't be tested in vivo using immunocompetent animal models due to potent xenograft responses. One can test the cells for safety, efficacy, and immunogenicity in patients, which will be time consuming, expensive, and potentially dangerous. Thus, a critical requirement for the translation of human stem cell-based therapy to the clinic is the development of robust pre-clinical animal models for evaluation of the efficacy, safety and importantly, immunogenicity of human cellular therapy. We propose to modify NOD-scid IL2rgnull (NSG) models of human diabetes and muscle disease to permit development of robust human immune systems. This will allow rapid preclinical evaluation of the function and immunogenicity of human SC-β cells and SC-myoblasts (and SC-derived cells in other models under development). We have documented that current models of NSG mice engrafted with human immune systems cannot fully reject allogeneic human ESCs or SC-endothelial cells, and in preliminary studies, cannot reject human SC-β cells. Our Scientific Premise is that the limited function of the engrafted human immune system in humanized NSG mice is due to lack of organized structure in secondary lymphoid tissues. We have observed that engraftment of newborn NSG mice with wildtype murine HSC will lead to the development of lymph nodes and robust immune responses, suggesting that a lack of species-specific factors impede lymphoid development in humanized NSG mice. In this multi-PI, multi-disciplinary team proposal, Aim 1 will generate novel NSG mouse strains of diabetes and muscular disease that express human specific factors that support lymphoid structure and human immune function. Aim 2 will engraft human SC-β cells and SC- myoblasts into our unique humanized mouse models of diabetes and muscle disease to test for function and immunogenicity. Aim 3 will provide these validated new strains to The Jackson Laboratory (JAX) Biorepository for worldwide distribution. Our proposal takes advantage of powerful new technologies for creating new models of humanized mice, and builds on our track record for generating, validating, and to date, sharing through the JAX distribution network of 17 novel models of humanized NSG mice since 2005. We believe that our innovative approaches, combined with our multi-disciplinary collaborative team will ensure the development of these much-needed preclinical models for regenerative medicine.