The astrocyte transcriptome in age and glaucoma: a comparative study of the optic nerve head, optic nerve proper and corpus callosum

年龄与青光眼的星形胶质细胞转录组：视神经乳头、视神经固有层和胼胝体的比较研究

• 批准号: 10707133
• 负责人: Daniel Sun
• 金额: $ 24.63万
• 依托单位: SCHEPENS EYE RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-30 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10707133
• 关键词: ATP binding cassette transporter 1; Affect; Age; Aging; Architecture; Astrocytes; Axon; Axonal Transport; Blindness; Blood Vessels; Brain; CSPG4 gene; Cell Separation; Cell Survival; Cells; Chronic; Comparative Study; Complement; Complex; Corpus Callosum; Data Set; Development; Disease; Disease Outcome; Distal; Extracellular Matrix; Functional disorder; Future; Galectin 3; Gene Expression; Gene Expression Profile; Genes; Genetic Transcription; Glaucoma; Glial Fibrillary Acidic Protein; Growth Factor; Head; Heterogeneity; Immunoprecipitation; Infiltration; Inflammation; Inflammatory; Injury; Knowledge; Label; Manufactured Baseball; Messenger RNA; Methods; Microglia; Microspheres; Modeling; Molecular; Morphology; Mus; Nerve; Neurons; Oligodendroglia; Optic Disk; Optic Nerve; Pathway interactions; Patients; Pattern; Phagocytosis; Pharmaceutical Preparations; Physiologic Intraocular Pressure; Play; Population; Positioning Attribute; Predisposition; Process; Property; Quantitative Reverse Transcriptase PCR; Reaction; Retinal Ganglion Cells; RiboTag; Ribosomes; Risk Factors; Role; Site; Spinal Cord; Techniques; Testing; Tissues; Transcript; Vimentin; Vision; Work; aged; alternative treatment; aquaporin 4; axon injury; axonal degeneration; cell injury; cell type; deprivation; differential expression; ganglion cell; genome-wide; knowledge base; mitochondrial metabolism; molecular phenotype; monocyte; myelination; nestin protein; neuroinflammation; novel; transcriptome; transcriptome sequencing; transcriptomic profiling; transcriptomics; white matter

Project Summary The underlying pathophysiological mechanisms in glaucomatous ganglion cell degeneration are still not clear. Because of this, current medications rely on modifying a major risk factor, which is intraocular pressure. However, lowering intraocular pressure alone is not always sufficient, and many patients continue to slowly lose vision. Identifying new mechanisms that are either harmful or beneficial for ganglion cell survival will benefit the development of alternative treatments for glaucoma. A leading hypothesis in the pathophysiology of glaucoma is that astrocytes within the optic nerve head (ONH) play an important role in the ganglion cell degeneration, albeit it is not clear what this role is. Astrocytes are a focus because (1) they populate the ONH, a site where early ganglion cell axon injury occurs, and (2) astrocytes in the ONH become highly “reactive”, a process that changes their morphology, function and molecular phenotype, but in the white matter of the CNS is not well understood. Although the field recognizes their importance, there is a fundamental lack of understanding of the properties of ONH astrocytes (and white matter astrocytes in general), what makes them different and what they are doing in disease. Studies to date have typically used whole ONH tissues and/or examined either a single or a small group of genes/pathways to better understand ONH astrocytes, but this is an incomplete picture. Here, we have combined whole transcriptome profiling with a novel ribotag strategy that allows us to isolate mRNAs specifically from astrocytes from different tissue regions. We will compare the astrocyte transcriptome from the unmyelinated ONH, the myelinated optic nerve proper and corpus callosum, in young and aged mice, and mice that have undergone chronic elevations in intraocular pressure. We hypothesize that ONH astrocytes are a molecularly distinct population compared to astrocytes in the more distal myelinated optic nerve proper, and that they have functional specializations associated with the fact that glaucomatous pathophysiological changes preferentially occur in the nerve head region. Age and elevations in intraocular pressure induces a unique transcriptional profile. Our previous morphological characterization and immunocytochemical labeling patterns strongly suggest that ONH astrocytes are indeed unique from those in the other regions. Our specific aims to test the hypothesis are: (1) investigate the transcriptional profile of astrocytes in the ONH, optic nerve proper and corpus callosum in both normal young (3 mths) and aged mice (12 mths), and (2) investigate the transcriptional profile of astrocytes from the ONH and optic nerve proper in young (3 mths) and aged mice (12 mths) following a chronic elevation in intraocular pressure. This exploratory proposal will provide an important knowledge base for future more focused hypothesis driven studies.

项目概要 青光眼神经节细胞变性的潜在病理生理机制仍不清楚 因此，目前的药物依赖于改变一个主要的危险因素，即眼压。 然而，仅仅降低眼压并不总是足够的，许多患者继续缓慢地降低眼压。 识别对神经节细胞生存有害或有益的新机制将导致失明。 有利于青光眼替代疗法的开发。 青光眼病理生理学的一个主要假设是视神经头内的星形胶质细胞 (ONH) 在神经节细胞变性中发挥重要作用，尽管尚不清楚其作用是什么。 成为焦点，因为 (1) 它们分布在 ONH，这是早期神经节细胞轴突损伤发生的部位，(2) ONH 中的星形胶质细胞变得高度“反应”，这一过程改变了它们的形态、功能和 分子表型，但在中枢神经系统白质中尚不清楚，尽管该领域已认识到。 尽管它们很重要，但人们对 ONH 星形胶质细胞（和白色星形胶质细胞）的特性根本缺乏了解。 星形胶质细胞），它们的不同之处以及它们迄今为止在疾病中的研究。 通常使用整个 ONH 组织和/或检查单个或一小群基因/途径 更好地了解 ONH 星形胶质细胞，但这并不是一个完整的图景。 在这里，我们将全转录组分析与新颖的核标签策略相结合，使我们能够 特异地从不同组织区域的星形胶质细胞中分离 mRNA，我们将比较星形胶质细胞。 来自年轻无髓鞘 ONH、有髓鞘视神经和胼胝体的转录组 以及老年小鼠和眼内压慢性升高的小鼠。 与更远端有髓鞘的星形胶质细胞相比，ONH 星形胶质细胞是分子上不同的群体 视神经本身，并且它们具有与青光眼相关的功能专门化 病理生理变化优先发生在神经头区域。年龄和眼内抬高。 压力诱导了独特的转录谱。 免疫细胞化学标记模式强烈表明 ONH 星形胶质细胞确实与 我们检验该假设的具体目标是：（1）研究转录谱。 正常年轻（3 个月）和老年小鼠 ONH、视神经固有层和胼胝体中的星形胶质细胞 （12 个月），以及（2）研究来自 ONH 和视神经本身的星形胶质细胞的转录谱 眼压慢性升高后的年轻小鼠（3 个月）和老年小鼠（12 个月）。 这一探索性提案将为未来更有针对性的假设提供重要的知识基础 驱动的研究。