Sahay Prity, Chakraborty Munmun, Rao Aparna
Hyderabad Eye Research Foundation (HERF), L.V. Prasad Eye Institute, Bhubaneswar, India.
KIIT School of Biotechnology, Bhubaneswar, India.
Front Mol Biosci. 2022 Apr 20;9:877250. doi: 10.3389/fmolb.2022.877250. eCollection 2022.
Pseudoexfoliation (PXF) is characterized by the accumulation of the exfoliative material in the eye and high rates of blindness if left untreated. Pseudoexfoliation glaucoma (PXG) is generally diagnosed too late due to its asymptomatic nature, necessitating the development of new effective screening tools for the early diagnosis of the disease. Thus, the increasing prevalence of this disease due to an aging population has demanded the identification of suitable biomarkers for the early detection of the disease or detection of the onset of glaucoma in the eyes with PXF. We applied a proteomics strategy based on a high-throughput screening method for the determination of proteins involving PXF and PXG pathogenesis. The lens capsule (LC), iris, and trabecular meshwork (TM) samples with PXF and PXG were taken by surgical trabeculectomy, and control samples were taken from the donor corneal buttons obtained from the institutional eye bank to characterize the proteome profile. Peptides from the LC were analyzed using liquid chromatography with tandem mass spectrometry (LC-MS/MS). The protein of interest and cytokine/chemokine profiles were verified using immunohistochemistry and the bio-plex kit assay, respectively. There were a total of 1433 proteins identified in the human LC, of which 27 proteins were overexpressed and eight proteins were underexpressed in PXG compared with PXF. Overexpressed proteins such as fibromodulin, decorin, lysyl oxidase homolog 1, collagen alpha-1(I) chain, collagen alpha-3(VI) chain, and biglycan were the major components of the extracellular matrix (ECM) proteins involved in cell-matrix interactions or ECM proteoglycans and the assembly and cross-linking of collagen fibrils. The ECM composition and homeostasis are altered in glaucoma. Thus, quantitative proteomics is a method to discover molecular markers in the eye. Monitoring these events can help evaluate disease progression in future studies.
假性剥脱（PXF）的特征是眼部有剥脱物质积聚，若不治疗，失明率很高。假性剥脱性青光眼（PXG）由于无症状，通常诊断过晚，因此需要开发新的有效筛查工具用于该病的早期诊断。所以，随着人口老龄化，这种疾病的患病率不断上升，这就要求识别合适的生物标志物，以便早期检测该疾病或检测PXF患者眼中青光眼的发病情况。我们应用了一种基于高通量筛选方法的蛋白质组学策略，以确定与PXF和PXG发病机制相关的蛋白质。通过手术小梁切除术获取PXF和PXG患者的晶状体囊（LC）、虹膜和小梁网（TM）样本，从机构眼库获得的供体角膜植片获取对照样本，以表征蛋白质组图谱。使用液相色谱串联质谱（LC-MS/MS）分析LC中的肽段。分别使用免疫组织化学和生物芯片试剂盒检测验证感兴趣的蛋白质和细胞因子/趋化因子谱。在人LC中总共鉴定出1433种蛋白质，与PXF相比，PXG中有27种蛋白质过表达，8种蛋白质低表达。过表达的蛋白质如纤调蛋白、核心蛋白聚糖、赖氨酰氧化酶同源物1、胶原蛋白α-1（I）链、胶原蛋白α-3（VI）链和双糖链蛋白聚糖是参与细胞-基质相互作用或ECM蛋白聚糖以及胶原纤维组装和交联过程的细胞外基质（ECM）蛋白质的主要成分。青光眼患者的ECM组成和稳态会发生改变。因此，定量蛋白质组学是一种在眼部发现分子标志物的方法。监测这些事件有助于在未来的研究中评估疾病进展。
