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： 青光眼是致盲的主要原因，了解青光眼的发病机制是一个重要的 视觉研究的目标。正如许多基于人口的研究所表明的那样， 非洲裔青光眼发病年龄较早，严重程度和风险增加， 与高加索人相比，失明（CC）。造成这些差异的根本机制 是未知的，但遗传影响可能有助于这些表型差异。TM是 在调节眼内压方面很重要，是目前最重要也是唯一的 青光眼发展的可改变风险因素。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 从非裔美国人（AA）和CC青光眼患者中获得的网状（TM）样本 在手术过程中表明SDPR（也称为Cavin 2）在AA中的表达显著较低， 与CC相比，这一发现引发了人们对它与生理学之间关系的探索 和TM的病理学。SDPR与其他洞穴蛋白和小窝蛋白一起， 与调节小窝，亚显微镜，质膜坑，这已被注意到， 在TM中丰富。小窝与广泛的细胞功能和疾病有关， 这些结构在眼睛中的作用尚未阐明。这些细胞器由 caveolin（CAV）和cavin蛋白。中国人CAV 1/CAV 2编码基因附近的基因变异 全基因组关联研究已经在青光眼患者中确定， 人口。第一个具体目的将评估SDPR基因/蛋白的表达，因为它涉及 AA和CC青光眼患者TM小窝的超微结构变化， 健康尸体供体，利用标准和免疫金标记透射电子显微镜， SDPR和CAV 1的显微镜检查。第二个具体目标是调查SDPR特定的短 发夹RNA（shRNA）介导的基因沉默在培养的人TM细胞，以评估 表型相关性在小窝，如我们在POAG TM的初步观察中所确定的。在 与自我报告的种族背景相反，所有青光眼标本和健康供体组织 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.这项创新的研究代表了SDPR/Cavin 2基因的首次探索， 表达，蛋白分布和超微结构的影响，并应提供 对POAG种族差异潜在的新机制的有价值的见解。知识 这些研究中获得的成果可能会导致新的治疗方法和预防/治疗策略 致盲性疾病