title: Proteomic profiling of human liver biopsies: hepatitis C virus-induced fibrosis and mitochondrial dysfunction.
abstract: Liver biopsies from hepatitis C virus (HCV)-infected patients offer the unique opportunity to study human liver biology and disease in vivo. However, the low protein yields associated with these small samples present a significant challenge for proteomic analysis. In this study we describe the application of an ultrasensitive proteomics platform for performing robust quantitative proteomic studies on microgram amounts of HCV-infected human liver tissue from 15 patients at different stages of fibrosis. A high-quality liver protein database containing 5,920 unique protein identifications supported high throughput quantitative studies using (16)O/(18)O stable isotope labeling in combination with the accurate mass and time (AMT) tag approach. A total of 1,641 liver biopsy proteins were quantified, and analysis of variance (ANOVA) identified 210 proteins exhibiting statistically significant differences associated with fibrosis stage. Hierarchical clustering showed that biopsies representative of later fibrosis stages (for example, Batts-Ludwig stages 3-4) exhibited a distinct protein expression profile, indicating an apparent down-regulation of many proteins when compared with samples from earlier fibrosis stages (for example, Batts-Ludwig stages 0-2). Functional analysis of these signature proteins suggests that impairment of key mitochondrial processes including fatty acid oxidation and oxidative phosphorylation, and response to oxidative stress and reactive oxygen species occurs during advanced stage 3 to 4 fibrosis.
  
  
  CONCLUSIONS
  The results reported here represent a significant advancement in clinical proteomics providing to our knowledge, the first demonstration of global proteomic alterations accompanying liver disease progression in patients chronically infected with HCV. Our findings contribute to a generally emerging theme associating oxidative stress and hepatic mitochondrial dysfunction with HCV pathogenesis.
authors.affiliation: Department of Microbiology, University of Washington, Seattle, WA 98195-8070, USA. ddiamond@u.washington.edu
authors.forename: Deborah L
authors.initials: DL
authors.lastname: Diamond
authors.forename: Jon M
authors.initials: JM
authors.lastname: Jacobs
authors.forename: Bryan
authors.initials: B
authors.lastname: Paeper
authors.forename: Sean C
authors.initials: SC
authors.lastname: Proll
authors.forename: Marina A
authors.initials: MA
authors.lastname: Gritsenko
authors.forename: Robert L
authors.initials: RL
authors.lastname: Carithers
authors.forename: Anne M
authors.initials: AM
authors.lastname: Larson
authors.forename: Matthew M
authors.initials: MM
authors.lastname: Yeh
authors.forename: David G
authors.initials: DG
authors.lastname: Camp
authors.forename: Richard D
authors.initials: RD
authors.lastname: Smith
authors.forename: Michael G
authors.initials: MG
authors.lastname: Katze
mesh_terms: D000328:Adult; D001706:Biopsy; D005260:Female; D016174:Hepacivirus; D006801:Humans; D008103:Liver Cirrhosis / Q000378:metabolism* / Q000473:pathology* / Q000821:virology; D008297:Male; D008875:Middle Aged; D008930:Mitochondria, Liver / Q000378:metabolism; D018384:Oxidative Stress; D011506:Proteins / Q000032:analysis*; D040901:Proteomics / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: D011506:Proteins