Proteomics of Urinary Vesicles Links Plakins and Complement to Polycystic Kidney Disease

作者：Mahdi Salih, Jeroen Demmers, Karel Bezstarosti, Wouter N. Leonhard, Monique Losekoot, Cees van Kooten, Ron T. Gansevoort, Dorien J.M. Peters, Robert Zietse, Ewout Jasper Hoorn · 发表于：Journal of the American Society of Nephrology · 年份：2016 · DOI：10.1681/asn.2015090994 · 被引用次数：73 · 研究领域：Extracellular vesicles in disease、Genetic and Kidney Cyst Diseases、Renal Diseases and Glomerulopathies

Novel therapies in autosomal dominant polycystic kidney disease (ADPKD) signal the need for markers of disease progression or response to therapy. This study aimed to identify disease-associated proteins in urinary extracellular vesicles (uEVs), which include exosomes, in patients with ADPKD. We performed quantitative proteomics on uEVs from healthy controls and patients with ADPKD using a labeled approach and then used a label-free approach with uEVs of different subjects (healthy controls versus patients with ADPKD versus patients with non-ADPKD CKD). In both experiments, 30 proteins were consistently more abundant (by two-fold or greater) in ADPKD-uEVs than in healthy- and CKD-uEVs. Of these proteins, we selected periplakin, envoplakin, villin-1, and complement C3 and C9 for confirmation because they were also significantly overrepresented in pathway analysis and were previously implicated in ADPKD pathogenesis. Immunoblotting confirmed higher abundances of the selected proteins in uEVs from three independent groups of patients with ADPKD. Whereas uEVs of young patients with ADPKD and preserved kidney function already had higher levels of complement, only uEVs of patients with advanced stages of ADPKD had increased levels of villin-1, periplakin, and envoplakin. Furthermore, all five proteins correlated positively with total kidney volume. Analysis in kidney tissue from mice with kidney-specific, tamoxifen-inducible Pkd1 deletion demonstrated higher expression in more seve...