title: A new workflow for proteomic analysis of urinary exosomes and assessment in cystinuria patients.
abstract: Cystinuria is a purely renal, rare genetic disease caused by mutations in cystine transporter genes and characterized by defective cystine reabsorption leading to kidney stones. In 14% of cases, patients undergo nephrectomy, but given the difficulty to predict the evolution of the disease, the identification of markers of kidney damage would improve the follow-up of patients with a higher risk. The aim of the present study is to develop a robust, reproducible, and noninvasive methodology for proteomic analysis of urinary exosomes using high resolution mass spectrometry. A clinical pilot study conducted on eight cystinuria patients versus 10 controls highlighted 165 proteins, of which 38 were up-regulated, that separate cystinuria patients from controls and further discriminate between severe and moderate forms of the disease. These proteins include markers of kidney injury, circulating proteins, and a neutrophil signature. Analysis of selected proteins by immunobloting, performed on six additional cystinuria patients, validated the mass spectrometry data. To our knowledge, this is the first successful proteomic study in cystinuria unmasking the potential role of inflammation in this disease. The workflow we have developed is applicable to investigate urinary exosomes in different renal diseases and to search for diagnostic/prognostic markers. Data are available via ProteomeXchange with identifier PXD001430.
authors.affiliation: INSERM U1151 and ‡Proteomics Platform Necker, PPN-3P5, SFR Necker, INSERM US24, Université Paris Descartes, Sorbonne Paris Cité , 75006 Paris, France.
authors.forename: Matthieu
authors.initials: M
authors.lastname: Bourderioux
authors.forename: Thao
authors.initials: T
authors.lastname: Nguyen-Khoa
authors.forename: Cerina
authors.initials: C
authors.lastname: Chhuon
authors.forename: Ludovic
authors.initials: L
authors.lastname: Jeanson
authors.forename: Danielle
authors.initials: D
authors.lastname: Tondelier
authors.forename: Marta
authors.initials: M
authors.lastname: Walczak
authors.forename: Mario
authors.initials: M
authors.lastname: Ollero
authors.forename: Soumeya
authors.initials: S
authors.lastname: Bekri
authors.forename: Bertrand
authors.initials: B
authors.lastname: Knebelmann
authors.forename: Estelle
authors.initials: E
authors.lastname: Escudier
authors.forename: Bernard
authors.initials: B
authors.lastname: Escudier
authors.forename: Aleksander
authors.initials: A
authors.lastname: Edelman
authors.forename: Ida Chiara
authors.initials: IC
authors.lastname: Guerrera
mesh_terms: D015415:Biomarkers / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D019295:Computational Biology; D003555:Cystinuria / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D055354:Exosomes / Q000378:metabolism*; D005260:Female; D005786:Gene Expression Regulation / Q000235:genetics*; D006801:Humans; D015151:Immunoblotting; D007525:Isoelectric Focusing; D008297:Male; D016253:Microscopy, Immunoelectron; D010865:Pilot Projects; D040901:Proteomics / Q000379:methods*; D016622:Silver Staining; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D015415:Biomarkers
keywords: biomarkers; cystinuria; exosomes; mass spectrometry; proteomics; urine