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 (RD) 疾病，例如色素性视网膜炎 (RP) 和莱伯先天性黑蒙 （LCA），导致感光（PR）细胞功能障碍和细胞死亡，最终导致失明。 虽然传统上不认为是视网膜变性，但阿尔茨海默病 (AD) 也表现出许多症状 遗传性视网膜变性的特征。在 AD 和遗传性感光器变性中，例如 LCA，激活的小胶质细胞与它们参与的营养不良区域密切相关 神经炎症过程。资助的家长补助金的最初目标旨在研究 AIPL1 突变导致早发性视网膜变性。虽然这无疑会揭露许多 关于这种使人衰弱的疾病的发病机制的令人着迷的发现，该模型缺乏一个关键要素 在体内发现，即小胶质细胞的存在。除了在视网膜层压过程中提供支持外， 发育过程中，小胶质细胞也参与神经炎症过程。在当前的提案中，我们将 通过将小胶质细胞纳入开发 3D 视网膜类器官，以我们现有的 LCA 模型为基础 这将更具生理性，并且能够像体内发生的那样，在 RD 过程中对垂死的细胞做出反应。 此外，我们将包括阿尔茨海默病患者衍生的 iPSC 来构建 3D 视网膜，以模拟基于 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，那么 会给我们一个共同的分子靶标。我们提出的小胶质细胞工作与 自从将小胶质细胞整合到我们现有的 3D 视网膜模型中以来，最初拨款的目标将增强而不是 不仅是我们正在进行的 LCA 研究，而且还将使我们能够扩展到基于 阿尔茨海默病。一个中心假设是人类 PSC 衍生出具有 LCA 和 AD 的 3D 视网膜类器官 相关突变将重现人类视网膜营养不良，导致 PR 丧失、小胶质细胞增多 AD 特征标记的激活和积累。单独研究 LCA 组织中的小胶质细胞是 一项值得注意的努力，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。该提案将连接两项创新技术； (1) 3D 视网膜类器官技术和 (2) 干细胞来源的小胶质细胞定向分化。鉴于 小胶质细胞参与许多神经退行性疾病该项目非常适合阐明 LCA 和 AD 的机制。这些研究不仅会给 RD 生物学带来新的见解 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隘的视网膜研究到更广泛的应用研究 神经退行性疾病，如阿尔茨海默病和 LCA。