Racial Disparities in glaucoma: Implications of SDPR/Cavin2 in human trabecular meshwork

青光眼的种族差异：SDPR/Cavin2 对人类小梁网的影响

• 批准号: 9808499
• 负责人: CARLA J SIEGFRIED
• 金额: $ 23.56万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-01 至 2021-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9808499
• 关键词: Affect; African; African American; Age; Antibodies; Biogenesis; Blindness; CAV1 gene; CAV2 gene; Cadaver; Caucasians; Caveolae; Caveolins; Cell Culture Techniques; Cell membrane; Cell physiology; Cells; Code; Cornea; DNA; Data Analyses; Development; Disease; Eye; Family; Functional disorder; Future; Gene Expression; Gene Expression Profiling; Gene Proteins; Gene Silencing; Genes; Genetic; Genotype; Glaucoma; Goals; Goniotomies; Health; High Prevalence; Human; Immunohistochemistry; In Situ Hybridization; In Vitro; Individual; Investigation; Knockout Mice; Knowledge; Label; Lasers; Lead; Link; Liquid substance; Mammalian Cell; Mediating; Microdissection; Microscopic; Modeling; Morphology; Operative Surgical Procedures; Organelles; Pathogenesis; Pathologic; Pathology; Pathway interactions; Patient Self-Report; Patients; Penetrating Keratoplasty; Phenotype; Physiologic Intraocular Pressure; Physiological; Physiology; Play; Polymerase Chain Reaction; Population; Population Study; Prevention; Primary Open Angle Glaucoma; Proteins; Public Health; Quantitative Evaluations; Race; Regulation; Research Proposals; Resistance; Risk; Risk Factors; Role; Serum; Severities; Specimen; Structure; Study of serum; Techniques; Therapeutic Intervention; Tissue Donors; Tissue-Specific Gene Expression; Tissues; Trabecular meshwork structure; Trabeculectomy; Transmission Electron Microscopy; Variant; Vision research; Visual impairment; Western Blotting; aqueous; base; caucasian American; caveolin 1; caveolin-2; cohort; deprivation; early onset; genetic variant; genome wide association study; genome-wide analysis; high intraocular pressure; improved; in vivo; innovation; insight; knock-down; modifiable risk; mouse model; novel; novel therapeutics; personalized therapeutic; pressure; prevent; protein distribution; protein expression; racial disparity; response; sex; small hairpin RNA; transcriptome sequencing; whole genome

Project Abstract: Understanding the pathogenesis of glaucoma, a leading cause of blindness, is an important goal of vision research. As indicated in numerous population-based studies, individuals of African descent develop glaucoma at an earlier age with increased severity and risk of blindness compared to Caucasians (CC). The fundamental mechanisms for these disparities are unknown, but genetic influences likely contribute to these phenotypic differences. The TM is important in the regulation of intraocular pressure, the most important and only current modifiable risk factor for glaucoma development. Differential gene expression analysis by RNA sequencing (RNAseq) and quantitative polymerase chain reaction (qPCR) compared trabecular meshwork (TM) specimens from African American (AA) and CC glaucoma patients obtained during surgery indicated SDPR (also known as Cavin2) had significantly lower expression in AA patients compared to CC. This finding led to explorations of how it may be related to physiology and pathology of the TM. SDPR, together with other cavins and caveolin proteins, is associated with regulation of caveolae, sub-microscopic, plasma membrane pits, which have been noted to be abundant in TM. Caveolae are linked to a wide range of cellular functions and disease, but the role of these structures in the eye has not been elucidated. These organelles are composed of caveolin (CAV) and cavin proteins. Gene variants near the CAV1/CAV2 encoding gene in genome wide association studies have been identified in glaucoma patients in select populations. The first specific aim will evaluate expression of SDPR gene/protein as it relates to the ultrastructural changes of caveolae in TM from both AA and CC glaucoma patients and healthy cadaver donors, utilizing both standard and immunogold labeling transmission electron microscopy for SDPR and CAV1. The second specific aim is to investigate SDPR-specific short hairpin RNA (shRNA)-mediated gene silencing in cultured human TM cells to evaluate phenotypic correlations in caveolae as identified in our preliminary observations in POAG TM. In contrast to self-reported racial background, all glaucoma specimens and healthy donor tissues will be confirmed by DNA genotyping, representing more accurate racial ancestry for data analysis. This innovative study represents the first exploration of SDPR/Cavin2 gene expression, protein distribution and ultrastructural effects in human TM and should provide valuable insights into a novel mechanism underlying racial disparities in POAG. The knowledge gained in these studies may lead to new therapies for and strategies to prevent/treat this blinding disease.

项目摘要： 了解青光眼的发病机理是失明的主要原因，是一个重要的 视力研究的目标。如许多基于人群的研究所示， 非洲血统在更早的年龄较早的年龄较早，严重程度增加和风险 与高加索人（CC）相比，失明。这些差异的基本机制 是未知的，但是遗传影响可能会导致这些表型差异。 TM是 对于眼内压的调节，最重要，最重要，唯一的电流很重要 青光眼发育的可修改风险因素。 RNA差异基因表达分析 测序（RNASEQ）和定量聚合酶链反应（QPCR）比较小梁 来自非裔美国人（AA）和CC青光眼患者的网格工作（TM）标本 手术期间表明SDPR（也称为Cavin2）在AA中的表达明显较低 与CC相比。这一发现导致探索它与生理的关系 和TM的病理。 SDPR与其他骑士和小窝蛋白蛋白一起关联 通过调节口腔，亚微镜，质膜坑的调节，已注意到 在TM中很丰富。 Caveolae与各种细胞功能和疾病有关，但 这些结构在眼中的作用尚未阐明。这些细胞器组成 小窝蛋白（CAV）和卡氏蛋白的蛋白质。 CAV1/CAV2附近的基因变体编码基因 在青光眼患者中已经确定了基因组广泛的研究 人群。第一个特定目的将评估SDPR基因/蛋白质的表达 AA和CC青光眼患者和 健康的尸体供体，同时使用标准和免疫金标签传输电子 SDPR和CAV1的显微镜。第二个具体目的是研究特定于SDPR的简短 发夹RNA（shRNA）介导的培养的人TM细胞中的基因沉默以评估 Caveolae中的表型相关性，如我们在POAG TM中的初步观察中所述。在 与自我报告的种族背景，所有青光眼标本和健康供体组织形成对比 DNA基因分型将证实，代表数据的更准确的种族血统 分析。这项创新研究代表了SDPR/Cavin2基因的第一次探索 人类TM中的表达，蛋白质分布和超微结构效应，应提供 对POAG中种族差异的一种新型机制的宝贵见解。知识 在这些研究中获得的可能会导致新的疗法和防止/治疗的策略 盲目疾​​病。