title: Characterization of glomerular diseases using proteomic analysis of laser capture microdissected glomeruli.
abstract: The application of molecular techniques to characterize clinical kidney biopsies has the potential to provide insights into glomerular diseases that cannot be revealed by traditional renal pathology. The present work is a proof-of-concept approach to test whether proteomic analysis of glomeruli isolated from clinical biopsies by laser capture microdissection can provide unique information regarding differentially expressed proteins relevant to disease pathogenesis. The proteomes of glomeruli isolated by laser capture microdissection from biopsies of normal kidneys (living-related donor kidneys) were compared with those from patients with diabetic nephropathy, lupus nephritis, and fibronectin glomerulopathy. Glomerular proteins were extracted, trypsin digested, and subjected to liquid chromatography-tandem mass spectrometry for identification and quantitation. Relative to normal glomeruli, all disease-associated glomeruli showed an increased presence of complement components, a marked decline in podocyte-associated proteins, and a decrease in proteins associated with cellular metabolism. Additionally, fibronectin glomerulopathy glomeruli differed from all the other glomeruli because of a significant accumulation of fibronectin and fibulin. This study demonstrates that our method acquires reproducible and quantitative proteomic information from laser capture microdissection isolates that can be used to characterize the molecular features of glomerular diseases.
authors.affiliation: Department of Pathology, The Ohio State University Medical Center, Columbus, OH, USA.
authors.forename: Anjali A
authors.initials: AA
authors.lastname: Satoskar
authors.forename: John P
authors.initials: JP
authors.lastname: Shapiro
authors.forename: Cherri N
authors.initials: CN
authors.lastname: Bott
authors.forename: Huijuan
authors.initials: H
authors.lastname: Song
authors.forename: Gyongyi M
authors.initials: GM
authors.lastname: Nadasdy
authors.forename: Sergey V
authors.initials: SV
authors.lastname: Brodsky
authors.forename: Lee A
authors.initials: LA
authors.lastname: Hebert
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Birmingham
authors.forename: Tibor
authors.initials: T
authors.lastname: Nadasdy
authors.forename: Michael A
authors.initials: MA
authors.lastname: Freitas
authors.forename: Brad H
authors.initials: BH
authors.lastname: Rovin
mesh_terms: D000293:Adolescent; D000328:Adult; D015415:Biomarkers / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D003928:Diabetic Nephropathies / Q000378:metabolism / Q000473:pathology; D005260:Female; D015432:Glomerulonephritis, Membranoproliferative / Q000378:metabolism / Q000473:pathology; D006801:Humans; D007674:Kidney Diseases / Q000175:diagnosis* / Q000378:metabolism / Q000473:pathology; D007678:Kidney Glomerulus / Q000737:chemistry / Q000378:metabolism / Q000473:pathology*; D061346:Laser Capture Microdissection / Q000379:methods*; D008181:Lupus Nephritis / Q000378:metabolism / Q000473:pathology; D008297:Male; D008565:Membrane Proteins / Q000737:chemistry / Q000378:metabolism*; D010375:Pedigree; D010449:Peptide Mapping; D040901:Proteomics / Q000379:methods*; D021241:Spectrometry, Mass, Electrospray Ionization
chemical_list: D015415:Biomarkers; D008565:Membrane Proteins