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（导致失明的主要原因）的发病机制非常重要 视觉研究的目标。正如大量基于人群的研究表明， 非洲人后裔患青光眼的年龄较早，且严重程度和风险增加 与白种人相比失明（CC）。这些差异的基本机制 尚不清楚，但遗传影响可能导致这些表型差异。 TM 是 对眼压调节很重要，是目前最重要且唯一的 青光眼发生的可改变危险因素。 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 从非裔美国人 (AA) 和 CC 青光眼患者获得的网状 (TM) 标本 手术期间表明 SDPR（也称为 Cavin2）在 AA 中的表达显着降低 患者与CC相比。这一发现引发了对其与生理学的关系的探索 和 TM 的病理学。 SDPR 与其他cavins 和caveolin 蛋白一起，与 具有对小凹、亚微观、质膜凹坑的调节，这些凹坑已被注意到 TM 丰富。小凹与多种细胞功能和疾病有关，但是 这些结构在眼睛中的作用尚未阐明。这些细胞器是由 Caveolin (CAV) 和 Cavin 蛋白。 CAV1/CAV2 编码基因附近的基因变异 已在选定的青光眼患者中进行了全基因组关联研究 人口。第一个具体目标将评估 SDPR 基因/蛋白质的表达，因为它与 AA 和 CC 青光眼患者 TM 中小凹的超微结构变化 健康尸体捐献者，利用标准和免疫金标记传输电子 SDPR 和 CAV1 的显微镜检查。第二个具体目标是调查 SDPR 特定的短期 在培养的人类 TM 细胞中进行发夹 RNA (shRNA) 介导的基因沉默以进行评估 正如我们在 POAG TM 中的初步观察所确定的，小凹中的表型相关性。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相比 将通过 DNA 基因分型进行确认，代表更准确的种族血统数据 分析。这项创新研究代表了对SDPR/Cavin2基因的首次探索 人类 TM 中的表达、蛋白质分布和超微结构效应，并应提供 对 POAG 种族差异背后的新机制提供了宝贵的见解。知识点 这些研究中获得的成果可能会带来新的疗法和预防/治疗策略 致盲疾病。