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: 10478218
• 负责人: Jason Stephen Meyer
• 金额: $ 38.86万
• 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/10478218
• 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）是视网膜的投影神经元，作为视网膜的连接 眼睛和大脑。在此角色中，他们允许将视觉信息传输到丘脑目标，并以 伤害或疾病损害这种途径，导致视力丧失或失明。神经胶质细胞，尤其是星形胶质细胞 小胶质细胞在视神经内发现与RGC相邻，在那里它们保持体内稳态条件 供RGC确保适当的健康和功能。相反，当神经炎症情况发生 星形胶质细胞和小胶质细胞被诱导采用反应性状态，导致RGC退化。 神经炎症与多种神经退行性疾病有关，但是 由于视神经中神经胶质反应性的高度局部性质，青光眼中的神经炎症是独特的 头部作为RGC轴突脱离眼睛，与青光眼沿RGC轴突的最初损伤部位相关。尽管 动物模型已经证明了神经胶质在神经元发展和变性中的重要性，重要 动物模型和人类患者之间存在差异，包括RGC的保存低以及 神经胶质的功能差异。因此，这些细胞相互作用的人类模型的发展 将进一步扩展我们对Glia如何为RGC提供支持的理解，以及神经胶质的反应 在神经炎症条件下，导致青光眼中RGC的变性。人多能茎 细胞（HPSC）可以作为视网膜发育和疾病研究的强大体外模型， 先前的研究表明，在体外对RGC神经退行性的能力进行了建模。但是，这些研究 尚未关注RGC的分隔性质，也不关注反应性神经胶质的影响 RGC在神经炎症条件下的轴突。因此，解决现有基于HPSC的缺点 青光眼的模型，为了更好地概括RGC和Glia之间的相互作用，当前应用 利用强大且可重现的体外模型来重现人性化神经胶质在人RGC上的空间相互作用 与青光眼中观察到的神经退行性表型有关的轴突。神经胶质与RGC之间的相互作用 隔室将在静态和反应性状态下进行分析，并且反应性的功能后果 RGC轴突上的神经胶质将在表型，转录和功能上进行评估，以确定 反应性神经胶质调节RGC神经变性。成功追求以下目标将利用 一个强大的微流体平台，用于分析RGC轴突，以包括神经胶质的神经炎症效应 并将提供机会进一步阐明人RGC中的基本神经退行性机制， 以及开发新的治疗方法来减慢或逆转神经变性。