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.

项目摘要 青光眼神经节细胞变性中的潜在病理生理机制仍然不是 清除。因此，当前的药物依赖于改变主要危险因素，即眼内压。 但是，仅降低眼压并不总是足够的，许多患者继续放慢速度 失去视力。确定对神经节细胞存活有害或有益的新机制将 受益于青光眼替代治疗的发展。 青光眼病理生理学中的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起重要作用，尽管尚不清楚这种作用是什么。星形胶质细胞 之所以成为重点，是因为（1）它们填充了ONH，一个早期神经节细胞轴突损伤的部位，（2） ONH中的星形胶质细胞变得高度“反应性”，这一过程改变了其形态，功能和 分子表型，但在中枢神经系统的白质中尚不清楚。虽然该领域识别 它们的重要性，根本缺乏对ONH星形胶质细胞（和白色的特性）的了解 总体而言，重要的是星形胶质细胞），是什么使它们与众不同以及它们在疾病中所做的事情。迄今为止的研究 通常使用整个ONH组织和/或检查了单个或一小部分基因/途径 更好地了解onh星形胶质细胞，但这是一个不完整的图片。 在这里，我们将整个转录组分析与一种新颖的Ribbonag策略相结合，使我们能够 从不同组织区域的星形胶质细胞中分离出mRNA。我们将比较星形胶质细胞 来自不髓鞘的ONH，髓鞘神经适当的转录组和callosum callosum callosum 和老年小鼠，以及在眼内压力上经历慢性升高的小鼠。我们假设这一点 与更远端的髓鞘中的星形胶质细胞相比 视觉神经适当，并且它们具有与青光眼相关的功能专长 病理生理变化优先发生在神经头区域。眼内的年龄和海拔 压力引起独特的转录轮廓。我们以前的形态学特征和 免疫细胞化学标记模式强烈表明，ONH星形胶质细胞确实是独一无二的 其他地区。我们测试假设的具体目的是：（1）研究 正常年轻（3个月）和老年小鼠的ONH，视神经正常和call体的星形胶质细胞 （12个月）和（2）研究适当的ANH和视神经的星形胶质细胞的转录曲线 眼内压力慢性升高后，年轻的（3个月）和老年小鼠（12个月）。 该探索性提案将为未来更集中的假设提供重要的知识基础 驱动的研究。