title: Proteomics of Urinary Vesicles Links Plakins and Complement to Polycystic Kidney Disease.
abstract: 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 severe stages of the disease and correlation with kidney weight for each protein of interest. In summary, proteomic analysis of uEVs identified plakins and complement as disease-associated proteins in ADPKD. These proteins are new candidates for evaluation as biomarkers or targets for therapy in ADPKD.
authors.affiliation: Department of Internal Medicine, Division of Nephrology & Transplantation, and.
authors.forename: Mahdi
authors.initials: M
authors.lastname: Salih
authors.affiliation: Proteomics Center, Erasmus Medical Center, Rotterdam, The Netherlands.
authors.forename: Jeroen A
authors.initials: JA
authors.lastname: Demmers
authors.affiliation: Proteomics Center, Erasmus Medical Center, Rotterdam, The Netherlands.
authors.forename: Karel
authors.initials: K
authors.lastname: Bezstarosti
authors.affiliation: Departments of Human Genetics.
authors.forename: Wouter N
authors.initials: WN
authors.lastname: Leonhard
authors.affiliation: Clinical Genetics, and.
authors.forename: Monique
authors.initials: M
authors.lastname: Losekoot
authors.affiliation: Nephrology, Leiden University Medical Center, Leiden, The Netherlands; and.
authors.forename: Cees
authors.initials: C
authors.lastname: van Kooten
authors.affiliation: Department of Nephrology, University Medical Center Groningen, Groningen, The Netherlands.
authors.forename: Ron T
authors.initials: RT
authors.lastname: Gansevoort
authors.affiliation: Departments of Human Genetics.
authors.forename: Dorien J M
authors.initials: DJ
authors.lastname: Peters
authors.affiliation: Department of Internal Medicine, Division of Nephrology & Transplantation, and.
authors.forename: Robert
authors.initials: R
authors.lastname: Zietse
authors.affiliation: Department of Internal Medicine, Division of Nephrology & Transplantation, and e.j.hoorn@erasmusmc.nl.
authors.forename: Ewout J
authors.initials: EJ
authors.lastname: Hoorn
mesh_terms: D000818:Animals; D003176:Complement C3 / Q000502:physiology*; D003186:Complement C9 / Q000502:physiology*; D000067128:Extracellular Vesicles; D006801:Humans; D051379:Mice; D051179:Plakins / Q000502:physiology*; D016891:Polycystic Kidney, Autosomal Dominant / Q000209:etiology*; D040901:Proteomics; D014556:Urine / Q000737:chemistry*
chemical_list: D003176:Complement C3; D003186:Complement C9; D051179:Plakins
keywords: ADPKD; complement; cytoskeleton; genetic renal disease