title: Mass spectrometry assays of plasma biomarkers to predict radiographic progression of knee osteoarthritis.
abstract: BACKGROUND
  Biomarkers to identify osteoarthritis (OA) patients at risk for disease progression are needed. As part of a proteomic analysis of knee synovial fluid from normal and OA patients, differentially expressed proteins were identified that could represent potential biomarkers for OA. This study aimed to use mass spectrometry assays to identify representative peptides from several proteins in synovial fluid and peripheral blood, and assess their levels as biomarkers of OA progression.
  
  
  METHODS
  Multiplexed high throughput selected reaction monitoring (SRM) assays were developed to measure tryptic peptides representative of 23 proteins in matched serum and synovial fluid samples from late OA subjects at the time of joint replacement. Subsequently plasma samples from the baseline visit of 173 subjects in an observational OA cohort were tested by SRM for peptides from nine of these proteins: afamin, clusterin, cartilage oligomeric matrix protein, hepatocyte growth factor, kallistatin, insulin-like growth factor binding protein, acid labile subunit, lubricin, lumican, and pigment epithelium-derived factor. Linear regression was used to determine the association between the peptide biomarker level at baseline and change in joint space width (ΔJSW) from baseline to 30 months, adjusting for age and sex.
  
  
  RESULTS
  In the matched cohort, 17 proteins could be identified in synovial fluid and 16 proteins were detected in serum. For the progression cohort, the average age was 62 and average ΔJSW over 30 months was 0.68 mm. A high correlation between different peptides from individual proteins was observed, indicating our assays correctly measured their target proteins. Peptides representative of clusterin, lumican and lubricin showed statistically significant associations with joint space narrowing after adjustment for age and sex. Partial R2 values showed clusterin FMETVAEK and lubricin LVEVNPK peptide biomarkers explains about 2 to 3% of the variability of ΔJSW, similar to that explained by age. A biomarker score combining normalized data for both lubricin and clusterin peptides increased the model R2 to 0.079.
  
  
  CONCLUSIONS
  Our results suggest that when combined, levels of peptides representative of clusterin and lubricin in plasma are as predictive of OA progression as age. Replication of these findings in other prospective OA cohorts is planned.
authors.forename: Susan Y
authors.initials: SY
authors.lastname: Ritter
authors.forename: Jamie
authors.initials: J
authors.lastname: Collins
authors.forename: Bryan
authors.initials: B
authors.lastname: Krastins
authors.forename: David
authors.initials: D
authors.lastname: Sarracino
authors.forename: Mary
authors.initials: M
authors.lastname: Lopez
authors.forename: Elena
authors.initials: E
authors.lastname: Losina
authors.forename: Antonios O
authors.initials: AO
authors.lastname: Aliprantis
mesh_terms: D000368:Aged; D000595:Amino Acid Sequence; D015415:Biomarkers / Q000032:analysis* / Q000097:blood / Q000378:metabolism; D011508:Chondroitin Sulfate Proteoglycans / Q000032:analysis / Q000097:blood / Q000378:metabolism; D051152:Clusterin / Q000032:analysis / Q000097:blood / Q000378:metabolism; D015331:Cohort Studies; D018450:Disease Progression; D005260:Female; D006023:Glycoproteins / Q000032:analysis / Q000097:blood / Q000378:metabolism; D006801:Humans; D007632:Keratan Sulfate / Q000032:analysis / Q000097:blood / Q000378:metabolism; D016014:Linear Models; D000071234:Lumican; D008297:Male; D013058:Mass Spectrometry / Q000379:methods*; D008875:Middle Aged; D020370:Osteoarthritis, Knee / Q000097:blood / Q000000981:diagnostic imaging* / Q000378:metabolism; D010455:Peptides / Q000032:analysis; D011379:Prognosis; D020543:Proteome / Q000032:analysis* / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D011859:Radiography; D013582:Synovial Fluid / Q000378:metabolism; D000099158:Proteoglycan 4
chemical_list: D015415:Biomarkers; D011508:Chondroitin Sulfate Proteoglycans; D051152:Clusterin; D006023:Glycoproteins; D007632:Keratan Sulfate; D000071234:Lumican; D010455:Peptides; D020543:Proteome; D000099158:Proteoglycan 4; C000604306:LUM protein, human