Humanized mice for investigating human stem cell-derived microglia in Alzheimers Disease

摘要 炎性小胶质细胞是神经炎症的关键调节因子，与阿尔茨海默病密切相关 疾病(AD)。由于小胶质细胞现在是阿尔茨海默病治疗干预的主要靶点，显然有必要 开发强大的临床前动物模型，允许在体内研究人类小胶质细胞，这将使 以小胶质细胞为基础的阿尔茨海默病新治疗方法的发展。 本补充提案是对NOT-AG-20-008的回应。这项补充提案的目标是 目的是开发新型免疫缺陷小鼠，作为研究人小胶质细胞在阿尔茨海默病中的资源。我们 已经组成了一个由小胶质细胞和神经退行性疾病方面的专业知识组成的多学科团队(Dr。 Schafer)以及在创建、验证和与科学研究人员共享人性化小鼠新模型方面的专业知识 社区(博士舒尔茨，布雷姆，格雷纳)。我们要求对我们正在进行的R24 OD026440进行补充资金 项目。R24项目的目标是开发一种免疫缺陷小鼠资源，用于人类研究 干细胞衍生的贝塔细胞和肌肉细胞。这一补充要求将实现我们搬家的目标 我们对人类干细胞及其后代在大脑、小胶质细胞和阿尔茨海默病环境中的分析。 人CSF1和IL34是小胶质细胞发育、生存和支持所必需的细胞因子 小胶质细胞的不同亚群。每个子集与AD的功能和关系尚不清楚。我们的科学 前提是我们的下一代NSG小鼠移植了人造血干细胞(HSC) 将发展出人类小胶质细胞亚群，这将使研究不同的小胶质细胞成为可能 亚群及其在阿尔茨海默病相关神经变性中的作用。我们开发了NOD-SCID IL2rgnull(NSG) 转基因表达人CSF1或人IL34的小鼠。这将使对人类小胶质细胞的研究 动态平衡状态。我们还从杰克逊的豪厄尔博士那里获得了NSG-TG(APP/PS1)菌株 转基因表达人/鼠嵌合前体蛋白的实验室(JAX) 淀粉样斑块。我们将建立NSG-TG(APP/PS1 hIL34)和NSG-TG(APP/PS1 hCSF1)小鼠进行研究 阿尔茨海默病的小胶质细胞。在目标1中，我们将验证和优化人小胶质细胞在人HSC中的植入。 移植NSG-TG(HIL34)、NSG-TG(HCSF1)，以及表达hIL34或hCSF1的NSG-TG(APP/PS1)小鼠。在……里面 目的2.确定人小胶质细胞的定位和免疫学特性，并测定其吞噬功能 与AD相关的神经退行性变的基础和炎症性质。 根据NOT-AG-20-008的要求，作为我们未来在AD中研究人类小胶质细胞的资源 将培育NSG-TG(HIL34 HCSF1)小鼠，并将这些小鼠与NSG-TG(APP/PS1)小鼠杂交，以便进行研究 人类小胶质细胞的两个子集分别处于稳态和疾病状态的单个受者。 这将为研究人类小胶质细胞和 它们在AD中的功能将通过日本宇宙航空研究开发机构的生物库随时提供给科学界。

ABSTRACT Inflammatory microglia are a critical regulator of neuroinflammation and are strongly linked to Alzheimer’s Disease (AD). As microglia are now a lead target for therapeutic intervention in AD, there is a clear need to develop robust pre-clinical animal models that permit investigation of human microglia in vivo, which will enable the development of new microglia-based treatments for AD. This supplemental proposal is in response to NOT-AG-20-008. The goal of this supplemental proposal is to develop novel immunodeficient mice as a resource for the study of human microglia in AD. We have formed a multi-disciplinary team consisting of expertise in microglia and neurodegenerative diseases (Dr. Schafer) and expertise in creating, validating, and sharing new models of humanized mice with the scientific community (Drs. Shultz, Brehm, Greiner). We request supplemental funds to our ongoing R24 OD026440 project. The goal of the R24 project is to develop a resource of immunodeficient mice for the study of human stem cell-derived beta cells and muscle cells. This supplemental request will accomplish our goal of moving our analyses of human stem cells and their progeny into the setting of the brain, microglia, and AD. Human CSF1 and IL34 are cytokines required for microglial development as well as survival and support different subsets of microglia. The function and relationship of each subset to AD is not known. Our Scientific Premise is that our next generation NSG mice transplanted with human hematopoietic stem cells (HSC) will develop subpopulations of human microglia that will allow the investigation of different microglial subsets and their role in AD-related neurodegeneration. We have developed NOD-scid IL2rgnull (NSG) mice that transgenically express human CSF1 or human IL34. This will allow study of human microglia in a homeostatic state. We have also obtained a NSG-Tg(APP/PS1) strain from Dr. Howell at The Jackson Laboratory (JAX) that transgenically expresses a chimeric mouse/human precursor protein of A and develops amyloid plaques. We will create NSG-Tg(APP/PS1 hIL34) and NSG-Tg(APP/PS1 hCSF1) mice to study microglia in AD. In Aim 1, we will validate and optimize the engraftment of human microglia in human HSC- engrafted NSG-Tg(hIL34), NSG-Tg(hCSF1), and in NSG-Tg(APP/PS1) mice expressing hIL34 or hCSF1. In Aim 2, we will define human microglia localization and immunological profile and determine their phagocytic and inflammatory properties basally and in the context of neurodegeneration associated with AD. As requested in NOT-AG-20-008, as a resource for our future investigation of human microglia in AD we will develop NSG-Tg(hIL34 hCSF1) mice and cross these mice with NSG-Tg(APP/PS1) mice to allow the study of both subsets of human microglia in a single recipient in a homeostatic and in a disease state, respectively. This will provide a critical resource of much needed validated platforms for investigating human microglia and their function in AD that will be readily available to the scientific community through the JAX Biorepository.