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.

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