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

用于研究阿尔茨海默病中人类干细胞衍生的小胶质细胞的人源化小鼠

• 批准号: 10120199
• 负责人: Michael Allen Brehm
• 金额: $ 42.08万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-15 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10120199
• 关键词: APP-PS1; Administrative Supplement; Alzheimer associated neurodegeneration; Alzheimer' s Disease; Alzheimer' s disease related dementia; Amyloid beta-Protein; Animal Model; Award; Beta Cell; Brain; Breeding; CSF1 gene; Characteristics; Clinical Treatment; Communities; Development; Disease; Engraftment; Etiology; Funding; Future; Gliosis; Goals; Grant; Hematopoietic stem cells; Histologic; Human; Immunodeficient Mouse; Immunologics; Individual; Inflammatory; Investigation; Lead; Link; Microglia; Modeling; Molecular; Monoclonal Antibody R24; Mus; Muscle Cells; Nerve Degeneration; Neurodegenerative Disorders; Pathogenesis; Pathologic; Pathology; Patients; Phagocytes; Phenotype; Population; Property; Protein Precursors; Reporting; Research Personnel; Research Support; Resources; Risk; Role; Sampling; Senile Plaques; Study models; Synapses; The Jackson Laboratory; Therapeutic; Therapeutic Intervention; TimeLine; Transgenic Mice; Transgenic Organisms; Transplantation; United States National Institutes of Health; Work; base; biobank; cohort; cytokine; experimental study; human stem cells; humanized mouse; immunogenicity; in vivo; induced pluripotent stem cell; macrophage; model development; mouse model; multidisciplinary; nervous system disorder; neuroinflammation; next generation; novel; pre-clinical; prevent; response; risk variant; targeted treatment; tau Proteins; therapeutic target; transcriptome sequencing

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.

摘要 炎症性小胶质细胞是神经炎症的关键调节因子，与阿尔茨海默氏症密切相关 疾病（AD）。由于小胶质细胞现在是AD治疗干预的主要靶点，因此显然需要 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 开发新的基于小胶质细胞的AD治疗方法。 本补充提案是对NOT-AG-20-008的回应。本补充提案的目的是 是开发新的免疫缺陷小鼠作为研究AD中人类小胶质细胞的资源。我们 已经形成了一个多学科的团队，包括在小胶质细胞和神经退行性疾病的专业知识（博士。 Schafer）和专业知识，创造，验证，并与科学家分享新的人源化小鼠模型。 社区（Drs. Shultz，Bürzm，Greiner）.我们请求对正在进行的R24 OD 026440提供补充资金 项目R24项目的目标是开发用于人类免疫缺陷研究的免疫缺陷小鼠资源。 干细胞衍生的β细胞和肌肉细胞。这一补充请求将实现我们的目标， 我们对人类干细胞及其后代在大脑、小胶质细胞和AD中的作用进行了分析。 人CSF 1和IL 34是小胶质细胞发育以及存活和支持所需的细胞因子 不同的小胶质细胞亚群每个子集与AD的功能和关系尚不清楚。我们的科学 假设我们的下一代NSG小鼠移植了人类造血干细胞（HSC）， 将开发人类小胶质细胞的亚群，这将允许研究不同的小胶质细胞， 亚群及其在AD相关神经变性中的作用。我们开发了NOD-scid IL 2 rgull（NSG） 转基因表达人CSF 1或人IL 34的小鼠。这将使人类小胶质细胞的研究在一个 稳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 JAX实验室（JAX），转基因表达嵌合小鼠/人前体蛋白的A β， 淀粉样斑块我们将建立NSG-Tg（APP/PS1 hIL 34）和NSG-Tg（APP/PS1 hCSF 1）小鼠进行研究 AD中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。 移植的NSG-Tg（hIL 34）、NSG-Tg（hCSF 1）和表达hIL 34或hCSF 1的NSG-Tg（APP/PS 1）小鼠中。在 目的2：明确人小胶质细胞的定位和免疫学特性， 和炎症性质的基础上，并在与AD相关的神经变性的背景下。 根据NOT-AG-20-008的要求，作为我们未来研究AD中人类小胶质细胞的资源，我们 将培育NSG-Tg（hIL 34 hCSF 1）小鼠，并将这些小鼠与NSG-Tg（APP/PS 1）小鼠杂交以进行研究 分别在稳态和疾病状态下的单个接受者中的两个人类小胶质细胞亚群。 这将为研究人类小胶质细胞提供急需的验证平台的关键资源， 它们在AD中的功能将通过JAX生物储存库随时提供给科学界。