title: Label-free quantitative urinary proteomics identifies the arginase pathway as a new player in congenital obstructive nephropathy.
abstract: Obstructive nephropathy is a frequently encountered situation in newborns. In previous studies, the urinary peptidome has been analyzed for the identification of clinically useful biomarkers of obstructive nephropathy. However, the urinary proteome has not been explored yet and should allow additional insight into the pathophysiology of the disease. We have analyzed the urinary proteome of newborns (n = 5/group) with obstructive nephropathy using label free quantitative nanoLC-MS/MS allowing the identification and quantification of 970 urinary proteins. We next focused on proteins exclusively regulated in severe obstructive nephropathy and identified Arginase 1 as a potential candidate molecule involved in the development of obstructive nephropathy, located at the crossroad of pro- and antifibrotic pathways. The reduced urinary abundance of Arginase 1 in obstructive nephropathy was verified in independent clinical samples using both Western blot and MRM analysis. These data were confirmed in situ in kidneys obtained from a mouse obstructive nephropathy model. In addition, we also observed increased expression of Arginase 2 and increased total arginase activity in obstructed mouse kidneys. mRNA expression analysis of the related arginase pathways indicated that the pro-fibrotic arginase-related pathway is activated during obstructive nephropathy. Taken together we have identified a new actor in the development of obstructive nephropathy in newborns using quantitative urinary proteomics and shown its involvement in an in vivo model of disease. The present study demonstrates the relevance of such a quantitative urinary proteomics approach with clinical samples for a better understanding of the pathophysiology and for the discovery of potential therapeutic targets.
authors.affiliation: From the ‡Centre National de la Recherche Scientifique, Institut de Pharmacologie et de Biologie Structurale, F-31077 Toulouse, France; §Université Paul Sabatier, Toulouse, France;
authors.forename: Chrystelle
authors.initials: C
authors.lastname: Lacroix
authors.affiliation: §Université Paul Sabatier, Toulouse, France; ¶Institut National de la Santé et de la Recherche Médicale (INSERM), U1048, Institut of Cardiovascular and Metabolic Disease, Toulouse, France;
authors.forename: Cécile
authors.initials: C
authors.lastname: Caubet
authors.affiliation: From the ‡Centre National de la Recherche Scientifique, Institut de Pharmacologie et de Biologie Structurale, F-31077 Toulouse, France; §Université Paul Sabatier, Toulouse, France;
authors.forename: Anne
authors.initials: A
authors.lastname: Gonzalez-de-Peredo
authors.affiliation: §Université Paul Sabatier, Toulouse, France; ¶Institut National de la Santé et de la Recherche Médicale (INSERM), U1048, Institut of Cardiovascular and Metabolic Disease, Toulouse, France;
authors.forename: Benjamin
authors.initials: B
authors.lastname: Breuil
authors.affiliation: From the ‡Centre National de la Recherche Scientifique, Institut de Pharmacologie et de Biologie Structurale, F-31077 Toulouse, France; §Université Paul Sabatier, Toulouse, France;
authors.forename: David
authors.initials: D
authors.lastname: Bouyssié
authors.affiliation: From the ‡Centre National de la Recherche Scientifique, Institut de Pharmacologie et de Biologie Structurale, F-31077 Toulouse, France; §Université Paul Sabatier, Toulouse, France;
authors.forename: Alexandre
authors.initials: A
authors.lastname: Stella
authors.affiliation: From the ‡Centre National de la Recherche Scientifique, Institut de Pharmacologie et de Biologie Structurale, F-31077 Toulouse, France; §Université Paul Sabatier, Toulouse, France;
authors.forename: Luc
authors.initials: L
authors.lastname: Garrigues
authors.affiliation: ‖Methodomics, Toulouse, France; **Institut de Mathématiques de Toulouse, UMR 5219, INSA de Toulouse, Université de Toulouse, 135 Avenue de Rangueil, F-31077 Toulouse, France;
authors.forename: Caroline
authors.initials: C
authors.lastname: Le Gall
authors.affiliation: §Université Paul Sabatier, Toulouse, France; ¶Institut National de la Santé et de la Recherche Médicale (INSERM), U1048, Institut of Cardiovascular and Metabolic Disease, Toulouse, France;
authors.forename: Anthony
authors.initials: A
authors.lastname: Raevel
authors.affiliation: §Université Paul Sabatier, Toulouse, France; ¶Institut National de la Santé et de la Recherche Médicale (INSERM), U1048, Institut of Cardiovascular and Metabolic Disease, Toulouse, France;
authors.forename: Angelique
authors.initials: A
authors.lastname: Massoubre
authors.affiliation: §Université Paul Sabatier, Toulouse, France; ¶Institut National de la Santé et de la Recherche Médicale (INSERM), U1048, Institut of Cardiovascular and Metabolic Disease, Toulouse, France;
authors.forename: Julie
authors.initials: J
authors.lastname: Klein
authors.affiliation: §Université Paul Sabatier, Toulouse, France; ¶Institut National de la Santé et de la Recherche Médicale (INSERM), U1048, Institut of Cardiovascular and Metabolic Disease, Toulouse, France; ‡‡Nephrology and Internal Medicine Department, University Children's Hospital, Toulouse, France;
authors.forename: Stéphane
authors.initials: S
authors.lastname: Decramer
authors.affiliation: §§Laboratoire de Biochimie Métabolique, IFB, CHU Purpan, and INSERM UMR 1037, CRCT CHU Rangueil, Toulouse, France.
authors.forename: Frédérique
authors.initials: F
authors.lastname: Sabourdy
authors.affiliation: §Université Paul Sabatier, Toulouse, France; ¶Institut National de la Santé et de la Recherche Médicale (INSERM), U1048, Institut of Cardiovascular and Metabolic Disease, Toulouse, France; ‡‡Nephrology and Internal Medicine Department, University Children's Hospital, Toulouse, France;
authors.forename: Flavio
authors.initials: F
authors.lastname: Bandin
authors.affiliation: From the ‡Centre National de la Recherche Scientifique, Institut de Pharmacologie et de Biologie Structurale, F-31077 Toulouse, France; §Université Paul Sabatier, Toulouse, France;
authors.forename: Odile
authors.initials: O
authors.lastname: Burlet-Schiltz
authors.affiliation: From the ‡Centre National de la Recherche Scientifique, Institut de Pharmacologie et de Biologie Structurale, F-31077 Toulouse, France; §Université Paul Sabatier, Toulouse, France;
authors.forename: Bernard
authors.initials: B
authors.lastname: Monsarrat
authors.affiliation: §Université Paul Sabatier, Toulouse, France; ¶Institut National de la Santé et de la Recherche Médicale (INSERM), U1048, Institut of Cardiovascular and Metabolic Disease, Toulouse, France; joost-peter.schanstra@inserm.fr.
authors.forename: Joost-Peter
authors.initials: JP
authors.lastname: Schanstra
authors.affiliation: §Université Paul Sabatier, Toulouse, France; ¶Institut National de la Santé et de la Recherche Médicale (INSERM), U1048, Institut of Cardiovascular and Metabolic Disease, Toulouse, France; joost-peter.schanstra@inserm.fr.
authors.forename: Jean-Loup
authors.initials: JL
authors.lastname: Bascands
mesh_terms: D000818:Animals; D001119:Arginase / Q000235:genetics / Q000652:urine*; D004195:Disease Models, Animal; D005786:Gene Expression Regulation; D006801:Humans; D006869:Hydronephrosis / Q000151:congenital / Q000473:pathology / Q000652:urine*; D007223:Infant; D007231:Infant, Newborn; D007668:Kidney / Q000378:metabolism* / Q000473:pathology; D008297:Male; D008810:Mice, Inbred C57BL; D020543:Proteome / Q000235:genetics / Q000378:metabolism*; D040901:Proteomics / Q000379:methods; D051437:Renal Insufficiency / Q000151:congenital / Q000473:pathology / Q000652:urine*; D015398:Signal Transduction
chemical_list: D020543:Proteome; C574619:ARG1 protein, human; C477631:ARG2 protein, human; D001119:Arginase