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： 视网膜退行性疾病(RD)，如视网膜色素变性(RP)和Leber先天性黑发 (LCA)，导致光感受器(PR)细胞功能障碍和细胞死亡，最终导致失明。 虽然不被经典地认为是视网膜变性，但阿尔茨海默病(AD)也表现出许多 遗传性视网膜变性的特征。在阿尔茨海默病和遗传性光感受器退化中， LCA，激活的小胶质细胞与营养不良区域密切相关，在那里它们参与 神经炎性过程。受资助的父母补助金的最初目的是研究 AIPL1突变在早发性视网膜变性发病中的作用虽然这无疑会揭开许多 关于这种令人衰弱的疾病的发病机制的有趣发现，这个模型缺乏一个关键元素， 在体内发现，即小胶质细胞的存在。除了为视网膜层压提供支持外， 在发育过程中，小胶质细胞也参与神经炎症过程。在目前的提案中，我们将 在我们现有的LCA模型的基础上，将小胶质细胞整合到正在开发的3D视网膜器质中 它将更具生理性，并能够像在体内那样对RD期间死亡的细胞做出反应。 此外，我们将包括阿尔茨海默病患者来源的IPSCs来构建3D视网膜来模拟基于AD的 营养不良。这一点非常重要，因为如果小胶质细胞在不同形式的RD中发挥类似的作用， 会给我们一个共同的分子目标。我们建议的小胶质细胞工作与 最初拨款的目的，因为将小胶质细胞整合到我们现有的3D视网膜模型中将不会增强 只有我们正在进行的LCA研究，也将使我们能够扩展到一个新的研发模式，基于 阿尔茨海默氏症。一个中心假设是，人类PSC衍生出具有LCA和AD的3D视网膜器官 相关突变将重现人类视网膜营养不良，导致PR丢失，小胶质细胞增加 阿尔茨海默病标志物的激活和积累。而研究LCA组织中的小胶质细胞本身就是 值得注意的是，以类似的方式探索AD组织将增加关于一般情况的重要信息 小胶质细胞对细胞应激和细胞死亡的反应。这项建议将连接两项创新技术；(1) 3D视网膜器官技术和(2)干细胞来源的小胶质细胞定向分化。给定 小胶质细胞在一系列神经退行性疾病中的作用这个项目非常适合阐明 LCA和AD的作用机制。这些研究不仅将为RD的生物学带来新的见解 疾病，但它也可能提供一个模型，以探索针对小胶质细胞的新疗法，这将使 我们的工作范围从狭隘的视网膜研究到更广泛适用的 神经退行性疾病，如阿尔茨海默病和LCA。