Establishing a human cellular model of retinal ganglion cell compartmentalization in neurodegeneration and neuroinflammation

建立神经变性和神经炎症中视网膜神经节细胞区室化的人类细胞模型

• 批准号: 10279666
• 负责人: Jason Stephen Meyer
• 金额: $ 42.16万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10279666
• 关键词: Address; Adopted; Affect; Animal Model; Astrocytes; Axon; Axonal Transport; Blindness; Brain; Cell Compartmentation; Cell Differentiation process; Cell model; Cells; Coculture Techniques; Dendrites; Development; Disease; Disease Progression; Ensure; Eye; Gene Expression Profile; Genetic Transcription; Glaucoma; Health; Human; In Vitro; Injury; Label; Lead; Length; Microfluidics; Microglia; Modeling; Morphology; Mutation; Nature; Nerve Degeneration; Neuraxis; Neurodegenerative Disorders; Neuroglia; Neurons; Optic Disk; Optic Nerve; Pathology; Pathway interactions; Patients; Phenotype; Play; Reproducibility; Retina; Retinal Ganglion Cells; Role; Signal Pathway; Site; Study models; System; Testing; Thalamic structure; cell type; human model; human pluripotent stem cell; in vitro Model; neurodegenerative phenotype; neuroinflammation; neuron development; neurotoxic; neurotoxicity; novel; novel therapeutic intervention; patch clamp; real-time images; retinal axon; retinal ganglion cell degeneration; retinal neuron; stem cell model; success; transcriptome sequencing; transmission process; visual information

SUMMARY Retinal ganglion cells (RGCs) are the projection neurons of the retina that serve as the connection between the eye and the brain. In this role, they allow for the transmission of visual information to thalamic targets, with damage to this pathway in injury or disease leading to vision loss or blindness. Glial cells, particularly astrocytes and microglia, are found adjacent to RGCs within the optic nerve, where they maintain homeostatic conditions for RGCs to ensure proper health and functionality. Conversely, neuroinflammatory conditions occur when astrocytes and microglia are induced to adopt a reactive state, leading to the degeneration of RGCs. Neuroinflammation has been associated with a variety of neurodegenerative diseases, but the pathology of neuroinflammation in glaucoma is unique due to the highly localized nature of glial reactivity in the optic nerve head as RGC axons exit the eye, correlated with the initial site of injury along RGC axons in glaucoma. While animal models have demonstrated the importance of glia in neuronal development and degeneration, important differences exist between animal models and human patients, including low conservation of RGCs as well as numerous functional differences in glia. As such, the development of a human model of these cellular interactions would further expand our understanding of how glia provide support for RGCs, as well as how glia respond during neuroinflammatory conditions leading to the degeneration of RGCs in glaucoma. Human pluripotent stem cells (hPSCs) can serve as powerful in vitro models for the study of retinal development and disease, with previous studies demonstrating the ability to model RGC neurodegeneration in vitro. However, these studies have not focused upon the compartmentalized nature of RGCs, nor how reactive glia disproportionally affect the axons of RGCs in neuroinflammatory conditions. Thus, to address the shortcomings of existing hPSC-based models of glaucoma and to better recapitulate interactions between RGCs and glia, the current application leverages a robust and reproducible in vitro model to recreate the spatial interactions of glia upon human RGC axons relevant to the neurodegenerative phenotypes observed in glaucoma. Interactions between glia and RGC compartments will be analyzed in quiescent and reactive states, and the functional consequences of reactive glia upon RGC axons will be assessed phenotypically, transcriptionally, and functionally to identify the extent to which reactive glia modulate RGC neurodegeneration. The successful pursuit of the following aims will leverage a powerful microfluidic platform for the analysis of RGC axons to include the neuroinflammatory effects of glia and will provide opportunities to further elucidate fundamental neurodegenerative mechanisms in human RGCs, as well as to develop novel therapeutic approaches to slow or reverse neurodegeneration.

概括 视网膜神经节细胞（RGC）是视网膜的投射神经元，充当视网膜和视网膜之间的连接。 眼睛和大脑。在这个角色中，它们允许将视觉信息传输到丘脑目标， 损伤或疾病导致该通路受损，导致视力丧失或失明。神经胶质细胞，特别是星形胶质细胞 和小胶质细胞，被发现与视神经内的 RGC 相邻，在那里它们维持稳态条件 RGC 确保适当的健康和功能。相反，当以下情况发生时，就会发生神经炎症状况： 星形胶质细胞和小胶质细胞被诱导进入反应状态，导致 RGC 退化。 神经炎症与多种神经退行性疾病有关，但其病理学 由于视神经中神经胶质反应性的高度局部性，青光眼中的神经炎症是独特的 RGC 轴突离开眼睛时的头部，与青光眼中 RGC 轴突的初始损伤部位相关。尽管 动物模型已经证明了神经胶质细胞在神经元发育和变性中的重要性，重要 动物模型和人类患者之间存在差异，包括 RGC 保守性低以及 神经胶质细胞的许多功能差异。因此，开发这些细胞相互作用的人类模型 将进一步加深我们对胶质细胞如何为 RGC 提供支持以及胶质细胞如何应对的理解 在导致青光眼中 RGC 退化的神经炎症条件下。人类多能干 细胞（hPSC）可以作为研究视网膜发育和疾病的强大体外模型， 之前的研究证明了在体外模拟 RGC 神经变性的能力。然而，这些研究 没有关注 RGC 的划分性质，也没有关注反应性神经胶质细胞如何不成比例地影响 神经炎症条件下 RGC 的轴突。因此，为了解决现有基于 hPSC 的缺点 青光眼模型并更好地概括 RGC 和神经胶质细胞之间的相互作用，当前的应用 利用稳健且可重复的体外模型来重建神经胶质细胞对人类 RGC 的空间相互作用 与青光眼中观察到的神经退行性表型相关的轴突。神经胶质细胞和 RGC 之间的相互作用 将在静态和反应状态下分析隔室，以及反应的功能后果 RGC 轴突上的神经胶质细胞将从表型、转录和功能上进行评估，以确定其程度 其中反应性神经胶质细胞调节 RGC 神经变性。成功实现以下目标将利用 强大的微流体平台，用于分析 RGC 轴突，包括神经胶质细胞的神经炎症效应 并将提供机会进一步阐明人类 RGC 的基本神经退行性机制， 以及开发新的治疗方法来减缓或逆转神经退行性变。