Glaucoma-connective tissue changes in optic nerve head

青光眼-视神经乳头结缔组织变化

基本信息

批准号：
7292721
负责人：
M. ROSARIO HERNANDEZ
金额：
$ 65.75万
依托单位：
NORTHWESTERN UNIVERSITY AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
1999
资助国家：
美国
起止时间：
1999-04-01 至 2009-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7292721
关键词：
4 hydroxynonenal Affect African American Age American Astrocytes Axon Binding Biochemical Bioinformatics Biological Assay Cell Adhesion Cell Line Cells Coculture Techniques Computer Analysis Connective Tissue Data Environment Gene Expression Gene Expression Profile Genes Glaucoma Growth Factor Human Hydrostatic Pressure Immunohistochemistry In Situ Hybridization In Vitro Inflammatory Laboratories Lead Lipopolysaccharides Maintenance Mechanical Stress Molecular Profiling Neurons Numbers Oligonucleotide Microarrays Optic Disk Oxidative Stress Pathway interactions Phenotype Physiological Population Predisposition Primary Open Angle Glaucoma Proteins Regulation Regulatory Pathway Research Retinal Ganglion Cells Reverse Transcriptase Polymerase Chain Reaction Risk Signal Pathway Signal Transduction Stimulus Stress Testing Tissue-Specific Gene Expression Tissues Variant base caucasian American cell type inhibitor/antagonist migration neuronal survival novel optic nerve disorder pressure response therapeutic target tool

项目摘要

DESCRIPTION (provided by applicant): Our long-term objective is to determine how astrocytes contribute to optic neuropathy in primary open angle glaucoma (POAG). Our laboratory and others have provided evidence that astrocytes are the cells responsible for many pathological changes in the glaucomatous optic nerve head (ONH). Our Specific Aims are: 1) To identify variations in expression of genes in ONH astrocytes of African Americans (AAs) compared with age-matched Caucasian Americans (CAs). 2) To identify mechanisms, which regulate the transition of quiescent astrocytes to reactive astrocytes and mechanisms that regulate specific functions associated with the reactive phenotype. 3) To identify specific genes which are expressed by reactive astrocytes in response to different stimuli. 4) To identify the mechanisms that maintains the reactive astrocyte in the glaucomatous ONH. 5) To characterize the molecular interactions between astrocytes and axons that may lead to a non-supportive microenvironment that damages remaining axons and underlies the progression of optic neuropathy. We will use oligonucleotide microarrays and computational analyses to determine whether AAs have a distinctive gene expression profile in ONH astrocytes compared to CAs. Quantitative RT-PCR, in situ hybridization, immunohistochemistry, and biochemical assays will confirm microarray data. We will use specific inhibitors and activity assays to identify the signaling pathways and determine the regulation of functions associated with the reactive astrocyte. Using oligonucleotide microarrays and computational analyses, we will determine whether exposure of astrocytes to elevated hydrostatic pressure, a proinflammatory stimulus, or to an oxidative stress induce differential gene expressions that are specific to each type of stress. By comparing expression profiles between reactive astrocytes exposed to elevated hydrostatic pressure and glaucomatous astrocytes grown in culture, we will identify gene expressions responsible for maintenance of the reactive phenotype. Using co-cultures of normal and glaucomatous ONH astrocytes and a neuronal cell line, we will determine whether gene products expressed by ONH astrocytes under hydrostatic pressure or in glaucoma affect positively or negatively neuronal survival in vitro. This research is significant because we seek to identify novel gene expression or regulatory pathways that contribute to the susceptibility of AAs to glaucomatous optic neuropathy. By determining how quiescent astrocytes become reactive astrocytes and how the reactive phenotype is maintained in the ONH, we may discover therapeutic targets to encourage specific reactive astrocyte responses that are supportive of axonal survival or to suppress those that are destructive. Our studies to characterize the molecular interactions of astrocytes and axons that may produce a non-supportive microenvironment for remaining healthy axons in the glaucomatous ONH will demonstrate mechanisms underlying progression of glaucomatous optic neuropathy.

描述（由申请人提供）：我们的长期目标是确定星形胶质细胞如何对原发性敞开角度青光眼（POAG）中的视神经病变贡献。我们的实验室和其他人提供了证据，表明星形胶质细胞是导致青光眼视神经头（ONH）许多病理变化的细胞。我们的具体目的是：1）与年龄匹配的高加索美国人（CAS）相比，非洲裔美国人（AAS）中基因表达的差异。 2）确定机制，这些机制调节静态星形胶质细胞向反应性星形胶质细胞和机制的过渡，这些机制调节与反应性表型相关的特定功能。 3）确定反应性星形胶质细胞对不同刺激的反应性星形胶质细胞表达的特定基因。 4）确定在青光眼ONH中保持反应性星形胶质细胞的机制。 5）表征星形胶质细胞和轴突之间的分子相互作用，这些相互作用可能导致非支持性的微环境，该环境会损害剩余的轴突并构成视神经病变的进展。我们将使用寡核苷酸微阵列和计算分析来确定与CAS相比，ONH星形胶质细胞中AAS是否具有独特的基因表达谱。定量RT-PCR，原位杂交，免疫组织化学和生化测定将确认微阵列数据。我们将使用特定的抑制剂和活性测定来识别信号通路，并确定与反应性星形胶质细胞相关的功能的调节。使用寡核苷酸的微阵列和计算分析，我们将确定星形胶质细胞暴露于静水压力升高，促炎性刺激或氧化应激是否会诱导与每种压力的特异性差异基因表达式。通过比较暴露于培养物中静水压力升高的反应性星形胶质细胞和青光眼星形胶质细胞之间的表达谱，我们将确定负责维持反应性表型的基因表达。使用正常和青光眼的ONH星形胶质细胞和神经元细胞系的共培养，我们将确定在静水压力下以ONH星形胶质细胞表达的基因产物或在青光眼中表达的基因产物是否在体外影响正面或负面神经元存活。这项研究很重要，因为我们试图鉴定出有助于AAS对青光眼视觉神经病的敏感性的新型基因表达或调节途径。通过确定静态星形胶质细胞如何成为反应性星形胶质细胞以及如何保持反应性表型在ONH中，我们可以发现治疗靶标，以鼓励支持轴突存活或抑制破坏性的特定反应性星形胶质细胞反应。我们的研究表征了可能产生非支撑性微环境的星形胶质细胞和轴突的分子相互作用，以在青光眼ONH中保持健康的轴突，这将证明glaucompomatios Optic Neuropathy进展的机制。