Characterization of glomerular diseases using proteomic analysis of laser capture microdissected glomeruli

• Published in: Modern pathology Vol. 25; no. 5; pp. 709 - 721
• Main Authors: Satoskar, Anjali A, Shapiro, John P, Bott, Cherri N, Song, Huijuan, Nadasdy, Gyongyi M, Brodsky, Sergey V, Hebert, Lee A, Birmingham, Daniel J, Nadasdy, Tibor, Freitas, Michael A, Rovin, Brad H
• Format: Journal Article
• Language: English
• Published: New York Elsevier Inc 01.05.2012; Nature Publishing Group US; Elsevier Limited
• Subjects: 631/1647/2067; 692/699/1585/2759/1419; 692/700/139/422; Adolescent; Adult; Biomarkers - metabolism; Biopsy; Chromatography, High Pressure Liquid; Diabetes; Diabetic Nephropathies - metabolism; Diabetic Nephropathies - pathology; Diabetic nephropathy; Female; glomerular disease; Glomerulonephritis, Membranoproliferative - metabolism; Glomerulonephritis, Membranoproliferative - pathology; Humans; kidney; Kidney diseases; Kidney Diseases - diagnosis; Kidney Diseases - metabolism; Kidney Diseases - pathology; Kidney Glomerulus - chemistry; Kidney Glomerulus - metabolism; Kidney Glomerulus - pathology; Laboratory Medicine; laser capture microdissection; Laser Capture Microdissection - methods; Lasers; Lupus; Lupus Nephritis - metabolism; Lupus Nephritis - pathology; Male; Mass spectrometry; Medicine; Medicine & Public Health; Membrane Proteins - chemistry; Membrane Proteins - metabolism; Nephrology; original-article; Pathogenesis; Pathology; Pedigree; Peptide Mapping; Proteins; Proteomics; Proteomics - methods; Scientific imaging; Spectrometry, Mass, Electrospray Ionization; United States > US; Ohio; kidney; mass spectrometry; glomerular disease; proteomics; laser capture microdissection; lupus
• ISSN: 0893-3952; 1530-0285; 1530-0285
• DOI: 10.1038/modpathol.2011.205

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.