Pluripotent Stem Cell Derived 3D Retinas for Studies of Early Onset Retinal Degeneration

多能干细胞衍生的 3D 视网膜用于研究早发性视网膜变性

• 批准号: 10287214
• 负责人: KARL J WAHLIN
• 金额: $ 39.47万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10287214
• 关键词: 3-Dimensional; Adoption; Aging; Alzheimer' s Disease; Alzheimer' s disease model; Alzheimer' s disease patient; Alzheimer' s disease risk; Amyloid; Amyloid beta-Protein; Biology; Blindness; Brain; Cell Death; Cells; Cellular Stress; Characteristics; Circadian Rhythms; Coculture Techniques; Color Visions; DLG4 gene; Deposition; Development; Disease; Disease model; Elements; Exhibits; Exposure to; Eye; Functional disorder; Funding; Future; Gene Expression Profiling; Genes; Genetic; Goals; Grant; Heredity; Histology; Homeostasis; Human; Immunohistochemistry; Impaired cognition; Individual; Inherited; Injury; Interferon Type II; Lead; Leber' s amaurosis; Lesion; Microglia; Modeling; Molecular Target; Mutation; Nature; Nerve Degeneration; Neuraxis; Neurodegenerative Disorders; Neurons; Organoids; Outcome; Parents; Pathogenesis; Pathogenicity; Pathway interactions; Photoreceptors; Physiological; Play; Pluripotent Stem Cells; Process; Reporter; Retina; Retinal Degeneration; Retinal Diseases; Retinal Dystrophy; Retinitis Pigmentosa; Role; Senile Plaques; Site; Structure; Study models; Synapses; TNF gene; Technology; Testing; Tissues; Translating; Uncertainty; Vacuum; Vision; Work; abeta oligomer; base; cell type; design; early onset; experience; fascinate; genome wide association study; human model; in vivo; induced pluripotent stem cell; innovative technologies; insight; macrophage; migration; mild cognitive impairment; neuroinflammation; new therapeutic target; novel therapeutics; parent grant; photoreceptor degeneration; presenilin-1; pressure; response; risk variant; sight restoration; stem cell differentiation; therapeutic target; three dimensional cell culture; transcriptome sequencing

Project Summary/Abstract: Retinal degenerative (RD) diseases, such as Retinitis pigmentosa (RP) and Leber congenital amaurosis (LCA), cause dysfunction and cell death of photoreceptor (PR) cells, ultimately leading to blindness. While not classically considered a retinal degeneration, Alzheimer’s disease (AD) also exhibits many hallmarks of hereditary retinal degenerations. In both AD and hereditary photoreceptor degenerations, like LCA, activated microglia are intimately associated with dystrophic regions where they participate in neuroinflammatory processes. The original AIMs of the funded parent grant sought to study the role of AIPL1 mutations in the onset of early onset retinal degeneration. While this will no doubt uncover many fascinating findings about the pathogenesis of this debilitating disease, this model is lacking a key element that is found in vivo, namely the presence of microglia. In addition to providing support for retinal lamination during development, microglia also participate in neuroinflammatory processes. In the current proposal we will build upon our existing LCA model by incorporating microglia into developing 3D retinal organoids which will be more physiologic and capable of responding to dying cells during RD as occurs in vivo. Furthermore, we will include Alzheimer’s patient derived iPSCs to build 3D retinas to model AD based dystrophy. This is extremely important because if microglia serve a similar role in different forms of RD this would give us a common molecular target. Our proposed microglia work is highly synergistic with the AIMs of the original grant since integrating microglia into our existing 3D retina model will enhance not only our ongoing LCA studies but also will enable us to branch out into a new model of RD based on Alzheimer’s disease. A central hypothesis is that human PSC derived 3D retina organoids with LCA and AD associated mutations will recapitulate human retinal dystrophy resulting in PR loss, increased microglial activation and accumulation of markers characteristic of AD. While studying microglia in LCA tissue by itself is a noteworthy endeavor, exploring AD tissue in a similar fashion will add important information on general responses of microglia to cell stress and cell death. This proposal will bridge two innovative technologies; (1) 3D retinal organoid technology and (2) directed differentiation of stem cell derived microglia. Given the involvement of microglia in a host of neurodegenerative disorders this project is highly appropriate to elucidate the mechanisms of both LCA and AD. Not only will these studies lead to new insights into the biology of RD disease, but it could also provide a model to explore new therapeutics targeting microglia which would expand the scope of our work from narrowly focused retinal studies to more broadly applicable studies of neurodegenerative disorders like Alzheimer’s disease and LCA.

项目摘要/摘要： 视网膜变性（RD）疾病，例如色素性视网膜炎（RP）和Leber先天性瘤 （LCA），引起光感受器（PR）细胞的功能障碍和细胞死亡，最终导致失明。 虽然不经典地认为残留变性，但阿尔茨海默氏病（AD）也表现出许多 遗传性视网膜变性的标志。在AD和遗传感受器变性中， LCA，活化的小胶质细胞与营养不良区域密切相关 神经炎症过程。资助的父母赠款意识的最初目的是研究 AIPL1突变在早期发作的常规变性开始时。虽然这无疑会发现很多 关于这种使人衰弱疾病的发病机理的引人入胜的发现，该模型缺乏一个关键因素 在体内发现，即小胶质细胞。除了为视网膜层压提供支持 开发，小胶质细胞也参与神经炎症过程。在当前的建议中，我们将 通过将小胶质细胞纳入开发3D视网膜器官来建立我们现有的LCA模型 与体内发生的情况一样，这将更加生理，能够在RD期间对垂死的细胞做出反应。 此外，我们将包括阿尔茨海默氏症患者衍生的IPSC来构建3D视网膜以基于AD的模型 营养不良。这非常重要，因为如果小胶质细胞在不同形式的RD中发挥相似的作用 将为我们提供一个共同的分子靶标。我们提出的小胶质细胞工作与 原始赠款的目的是因为将小胶质细胞整合到我们现有的3D视网膜模型中不会增强 只有我们正在进行的LCA研究，但也将使我们能够基于 阿尔茨海默氏病。一个中心假设是，人PSC衍生出具有LCA和AD的3D视网膜器官 相关的突变将概括人类视网膜营养不良，导致PR损失，小胶质细胞增加 AD特征的标记的激活和积累。同时研究LCA组织中的小胶质细胞是 值得注意的努力，以类似的方式探索广告组织将添加有关一般信息的重要信息 小胶质细胞对细胞应激和细胞死亡的反应。该提议将桥接两种创新技术。 （1） 3D视网膜器官技术和（2）定向干细胞衍生的小胶质细胞的分化。鉴于 小胶质细胞参与许多神经退行性疾病，此项目非常适合阐明 LCA和AD的机制。这些研究不仅会导致对RD生物学的新见解 疾病，但它也可以提供一个模型来探索针对小胶质细胞的新疗法，这将扩大 我们的工作范围从狭oppo的视网膜研究到更广泛的适用研究 神经退行性疾病，例如阿尔茨海默氏病和LCA。