Development of a stem cell platform for the treatment of neurodegenerativemicrogliopathies

开发用于治疗神经退行性小胶质细胞病的干细胞平台

• 批准号: 10323423
• 负责人: Sunil Gandhi
• 金额: $ 39.9万
• 依托单位: NOVOGLIA, INC.
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-30 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10323423
• 关键词: Address; Adult; Alzheimer' s Disease; Axon; Behavioral; Biotechnology; Bone Marrow; Brain; Brain Pathology; CSF1R gene; Cell Count; Cell Proliferation; Cell Therapy; Cells; Clinical Trials; Cognitive; Data; Deep Cervical Lymph Node; Development; Disease; Disease model; Enhancers; Flow Cytometry; Frontotemporal Dementia; Future; Genes; Genetic Models; Glial Fibrillary Acidic Protein; Goals; Human; Immune; Immunohistochemistry; Industry; Inherited; Injections; Lead; Leukoencephalopathy; Licensing; Light; Liver; Location; Microglia; Modeling; Mus; Mutation; Neoplasms; Nerve Degeneration; Neurodegenerative Disorders; Neuroglia; Organ; PGRN gene; Pathologic; Pathology; Patients; Peripheral; Pharmacology and Toxicology; Phase; Pigments; Process; Regulatory Element; Research; Research Personnel; Risk; Safety; Single Nucleotide Polymorphism; Spleen; Stains; Stem Cell Development; Stem cell transplant; Symptoms; Technology; Testing; Therapeutic; Therapeutic Uses; Tissues; Transplantation; Treatment Efficacy; Undifferentiated; Work; astrogliosis; brain cell; brain tissue; calcification; clinically relevant; commercialization; confocal imaging; drug discovery; effective therapy; experimental study; genome wide association study; human pluripotent stem cell; induced pluripotent stem cell; insight; leukodystrophy; loss of function mutation; motor deficit; mouse genetics; mouse model; nervous system disorder; neurofilament; neuropathology; non-Native; osteopontin; phase 2 study; post-transplant; stem cell biomarkers; stem cell proliferation; stem cell therapy; stem cells; therapeutic evaluation; transplantation therapy

Project Summary We have developed a platform technology that uses induced human pluripotent stem cells (iPSCs) to generate genetically corrected microglia at scale and with high purity for transplantation. In this study, we will demonstrate the safety and therapeutic efficacy of our microglia transplantation technology using a mouse genetic model of adult-onset leukoencephalopathy with axonal spheroids & pigmented glia (ALSP), a fatal disease with no effective treatments. The three overlapping aims to be investigated in this Phase I project are: Aim 1: Optimize microglial transplantation delivery parameters in a mouse genetic model of microglial insufficiency. We have preliminary data showing that our iPSC-derived microglia can repopulate the brains of microglia-deficient mice. We will test three different microglia injection locations, and after determining the optimal injection location using whole brain clearing and quantification, we will then evaluate different numbers of cells at this optimized location. Aim 2: Determine the safety of microglial transplantation in the mouse model of microglial insufficiency. To assess the safety of our microglial repopulation, we will determine the percentage of cells that express non- microglial markers, examine brain tissue using immunohistochemistry for markers of undifferentiated iPSCs and proliferative cell markers, and determine the presence of any non-native human cells in relevant peripheral tissues. Aim 3: Demonstrate the efficacy of microglial transplantation on reduction of pathological ALSP hallmarks in the mouse model brain. Brain calcification, axonal spheroids, and astrogliosis are hallmarks of ALSP in human cases, as well as the mouse model. We have preliminary data demonstrating that microglial transplantation reduces these hallmark pathologies. Our delivery parameters from Aim 1 will be used for microglial transplantation. We will assess pathology reduction in the Aim 1 optimized delivery paradigm mice using confocal imaging and quantification of: osteopontin and Alazarin Red staining to identify calcified tissue, neurofilament light chain staining to identify axonal spheroids, and GFAP staining to identify astrogliosis. We expect a > 30% reduction in all of these pathologies with microglial transplantation. Our Phase I results will provide proof-of-concept data to support the development of iPSC-derived microglial cell therapy for ALSP and related neurodegenerative microgliopathies.

项目摘要 我们已经开发了一种平台技术，该技术使用诱导的人类多能干细胞（iPSC）来产生 大规模遗传校正的小胶质细胞，并且具有用于移植的高纯度。在这项研究中，我们将证明 我们的小胶质细胞移植技术的安全性和治疗效果，使用小鼠遗传模型， 伴有轴突球状体和色素胶质细胞的成人型白质脑病（ALSP），一种致命的疾病， 有效的治疗。在第一阶段项目中将调查的三个重叠目标是： 目的1：在小胶质细胞遗传模型中优化小胶质细胞移植递送参数 不足我们有初步的数据显示，我们的iPSC衍生的小胶质细胞可以重新填充大脑， 小胶质细胞缺陷小鼠。我们将测试三种不同的小胶质细胞注射位置，并在确定 使用全脑清除和量化的最佳注射位置，然后我们将评估不同的数字 在这个最佳位置的细胞。 目的2：确定小胶质细胞移植在小胶质细胞功能不全小鼠模型中的安全性。到 为了评估我们的小胶质细胞再生的安全性，我们将确定表达非- 小胶质细胞标记物，使用免疫组织化学检查脑组织中未分化的iPSC的标记物， 增殖细胞标志物，并确定相关外周血中任何非天然人细胞的存在。 组织中 目的3：证明小胶质细胞移植在减少脑缺血/再灌注损伤中病理性ALSP标志物方面的功效。 老鼠的大脑模型脑钙化、轴索球状体和星形胶质细胞增生是人类ALSP的特征 例，以及小鼠模型。我们有初步数据表明小胶质细胞移植 减少了这些标志性的病理。我们来自Aim 1的递送参数将用于小胶质细胞 移植我们将使用共聚焦显微镜评估Aim 1优化递送范式小鼠中的病理学减少。 成像和定量：骨桥蛋白和阿拉扎林红染色，以识别钙化组织、神经丝 轻链染色以鉴定轴突球状体，GFAP染色以鉴定星形胶质细胞增生。我们预计， 小胶质细胞移植减少了所有这些病理。 我们的I期结果将提供概念验证数据，以支持iPSC衍生的小胶质细胞的开发。 治疗ALSP和相关的神经退行性小胶质细胞病。