Profiling of vitreous proteomes from proliferative diabetic retinopathy and nondiabetic patients

Abstract

Diabetes can lead to serious microvascular complications like proliferative diabetic retinopathy (PDR), which is the leading cause of blindness in adults. The proteomic changes that occur during PDR cannot be measured in the human retina for ethical reasons, but could be reflected by proteomic changes in vitreous humor. Thus, we considered that comparisons between the proteome profiles of the vitreous humors of PDR and nondiabetic controls could lead to the discovery of novel pathogenic proteins and clinical biomarkers. In this study, the authors used several proteomic methods to comprehensively examine vitreous humor proteomes of PDR patients and nondiabetic controls. These methods included immunoaffinity subtraction (IS)/2-DE/ MALDI-MS, nano-LC-MALDI-MS/MS, and nano-LC-ESI-MS/MS. The identified proteins were subjected to the Trans-Proteomic Pipeline validation process. Resultantly, 531 proteins were identified, i.e., 415 and 346 proteins were identified in PDR and nondiabetic control vitreous humor samples, respectively, and of these 531 proteins, 240 were identified for the first time in this study. The PDR vitreous proteome was also found to contain many proteins possibly involved in the pathogenesis of PDR. The proteins described provide the most comprehensive proteome listing in the vitreous humor samples of PDR and nondiabetic control patients.

Characterisation of the vitreous proteome in proliferative diabetic retinopathy

“…Carbonic anhydrase (CA) has been identified by proteomic analysis in the vitreous humour from PDR patients; CA production was increased in the vitreous of diabetic retinopathy patients compared with the controls [7,8]. In this study, we have confirmed this finding.…”

“…Previous studies have identified some proteins that are differentially expressed in the vitreous of PDR patients. Many of the proteins are involved in such processes as angiogenesis, cellular proliferation, acute phase response, vascular permeability changes, and oxygen-induced vessel loss [7,13]. These studies involved the use of vitreous from non-diabetic patients with macular hole (MH) or macular epiretinal membrane (MEM) as control groups.…”

Systems pathology analysis identifies neurodegenerative nature of age‐related vitreoretinal interface diseases

“…Mass spectrometry (MS)‐based quantitative proteomics provides a means for the determination of global proteome changes at the tissue and cellular levels, enabling a molecular level characterization of the pathophysiologies of complex eye disorders. Currently, most proteomic studies characterizing disease‐induced vitreous proteome changes have focused on proliferative and nonproliferative diabetic retinopathies (Kim et al, 2007; Loukovaara et al, 2015; Wang, Feng, Hu, Xie, & Wang, 2013), proliferative vitreoretinopathy (Garweg, Tappeneiner, & Halberstadt, 2013; Mitry, Fleck, Wright, Campbell, & Charteris, 2010), and age‐related macular degeneration (AMD; Koss et al, 2014), whereas iERM and MH remain less‐studied (Mandal et al, 2013; Pollreisz et al, 2013; Yu et al, 2014; Zhang et al, 2017). …”

Characterization of the Vitreous Proteome in Diabetes without Diabetic Retinopathy and Diabetes with Proliferative Diabetic Retinopathy

“…5 A recent study using pooled vitreous samples and protein identification based on at least one peptide match has further expanded the list of vitreous proteins. 13 In this report, we have expanded the number of independent vitreous samples analyzed and have compiled and compared results generated from X!Tandem and SEQUEST algorithms. This analysis resulted in the identification of 252 proteins, which was based on at least 2 unique peptide matches and confirmation in independent vitreous samples.…”