title: Urine proteome analysis in Dent's disease shows high selective changes potentially involved in chronic renal damage.
abstract: Definition of the urinary protein composition would represent a potential tool for diagnosis in many clinical conditions. The use of new proteomic technologies allows detection of genetic and post-trasductional variants that increase sensitivity of the approach but complicates comparison within a heterogeneous patient population. Overall, this limits research of urinary biomarkers. Studying monogenic diseases are useful models to address this issue since genetic variability is reduced among first- and second-degree relatives of the same family. We applied this concept to Dent's disease, a monogenic condition characterised by low-molecular-weight proteinuria that is inherited following an X-linked trait. Results are presented here on a combined proteomic approach (LC-mass spectrometry, Western blot and zymograms for proteases and inhibitors) to characterise urine proteins in a large family (18 members, 6 hemizygous patients, 6 carrier females, and 6 normals) with Dent's diseases due to the 1070G>T mutation of the CLCN5. Gene ontology analysis on more than 1000 proteins showed that several clusters of proteins characterised urine of affected patients compared to carrier females and normal subjects: proteins involved in extracellular matrix remodelling were the major group. Specific analysis on metalloproteases and their inhibitors underscored unexpected mechanisms potentially involved in renal fibrosis.
  
  
  CONCLUSIONS
  Studying with new-generation techniques for proteomic analysis of the members of a large family with Dent's disease sharing the same molecular defect allowed highly repetitive results that justify conclusions. Identification in urine of proteins actively involved in interstitial matrix remodelling poses the question of active anti-fibrotic drugs in Dent's patients.
authors.affiliation: Laboratory on Physiopathology of Uremia, Istituto Giannina Gaslini, Genova, Italy.
authors.forename: Laura
authors.initials: L
authors.lastname: Santucci
authors.affiliation: Laboratory on Physiopathology of Uremia, Istituto Giannina Gaslini, Genova, Italy.
authors.forename: Giovanni
authors.initials: G
authors.lastname: Candiano
authors.affiliation: Laboratory of Histomorphology and Molecular Biology of the Kidney, Division of Nephrology, University of Padova, Padova, Italy.
authors.forename: Franca
authors.initials: F
authors.lastname: Anglani
authors.affiliation: Laboratory on Physiopathology of Uremia, Istituto Giannina Gaslini, Genova, Italy.
authors.forename: Maurizio
authors.initials: M
authors.lastname: Bruschi
authors.affiliation: Laboratory of Histomorphology and Molecular Biology of the Kidney, Division of Nephrology, University of Padova, Padova, Italy.
authors.forename: Enrica
authors.initials: E
authors.lastname: Tosetto
authors.affiliation: Laboratory of Histomorphology and Molecular Biology of the Kidney, Division of Nephrology, University of Padova, Padova, Italy.
authors.forename: Daniela
authors.initials: D
authors.lastname: Cremasco
authors.affiliation: Nephrology, Dialysis, Transplantation Unit, Azienda Ospedaliera-University of Padova, Padova, Italy.
authors.forename: Luisa
authors.initials: L
authors.lastname: Murer
authors.affiliation: Proteomics and Mass Spectrometry Laboratory, Istituto per il Sistema Produzione Animale in Ambiente Mediterraneo (ISPAAM) National Research Council (CNR), Napoli, Italy.
authors.forename: Chiara
authors.initials: C
authors.lastname: D'Ambrosio
authors.affiliation: Proteomics and Mass Spectrometry Laboratory, Istituto per il Sistema Produzione Animale in Ambiente Mediterraneo (ISPAAM) National Research Council (CNR), Napoli, Italy.
authors.forename: Andrea
authors.initials: A
authors.lastname: Scaloni
authors.affiliation: Laboratory of Mass Spectrometry - Core Facility, Istituto Giannina Gaslini, Genova, Italy.
authors.forename: Andrea
authors.initials: A
authors.lastname: Petretto
authors.affiliation: Nephrology, Dialysis and Transplantation Unit, Istituto Giannina Gaslini, Genova, Italy.
authors.forename: Gianluca
authors.initials: G
authors.lastname: Caridi
authors.affiliation: Nephrology, Dialysis and Transplantation Unit, Istituto Giannina Gaslini, Genova, Italy.
authors.forename: Roberta
authors.initials: R
authors.lastname: Rossi
authors.affiliation: Nephrology, Dialysis and Transplantation Unit, Istituto Giannina Gaslini, Genova, Italy.
authors.forename: Alice
authors.initials: A
authors.lastname: Bonanni
authors.affiliation: Nephrology, Dialysis and Transplantation Unit, Istituto Giannina Gaslini, Genova, Italy; Laboratory on Physiopathology of Uremia, Istituto Giannina Gaslini, Genova, Italy. Electronic address: GMarcoGhiggeri@ospedale-gaslini.ge.it.
authors.forename: Gian Marco
authors.initials: GM
authors.lastname: Ghiggeri
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D015415:Biomarkers / Q000652:urine; D002648:Child; D057973:Dent Disease / Q000652:urine*; D005109:Extracellular Matrix / Q000378:metabolism; D005192:Family Health; D005260:Female; D005355:Fibrosis / Q000473:pathology; D014644:Genetic Variation; D006579:Heterozygote; D006801:Humans; D007676:Kidney Failure, Chronic / Q000652:urine*; D008297:Male; D008875:Middle Aged; D009154:Mutation; D010375:Pedigree; D011507:Proteinuria / Q000473:pathology / Q000652:urine*; D020543:Proteome / Q000378:metabolism*; D040901:Proteomics; D055815:Young Adult
chemical_list: D015415:Biomarkers; D020543:Proteome
keywords: CLCN5 gene; Chronic renal failure; Dent's disease; Fibrosis; Metalloproteases; TIMPs