title: Overexpression of MMP13 in human osteoarthritic cartilage is associated with the SMAD-independent TGF-β signalling pathway.
abstract: BACKGROUND
  In vitro and animal model of osteoarthritis (OA) studies suggest that TGF-β signalling is involved in OA, but human data is limited. We undertook this study to elucidate the role of TGF-β signalling pathway in OA by comparing the expression levels of TGFB1 and BMP2 as ligands, SMAD3 as an intracellular mediator, and MMP13 as a targeted gene between human osteoarthritic and healthy cartilage.
  
  
  METHODS
  Human cartilage samples were collected from patients undergoing total hip/knee joint replacement surgery due to primary OA or hip fractures as controls. RNA was extracted from the cartilage tissues. Real-time quantitative PCR was performed to measure gene expression. Mann-Whitney test was utilized to compare the expression levels of TGFB1, BMP2, SMAD3 and MMP13 in human cartilage between OA cases and controls. Spearman's rank correlation coefficient (rho) was calculated to examine the correlation between the expression levels of the four genes studied and non-parametric regression was used to adjust for covariates.
  
  
  RESULTS
  A total of 32 OA cases (25 hip OA and 7 knee OA) and 21 healthy controls were included. The expression of TGFB1, SMAD3, and MMP13 were on average 70%, 46%, and 355% higher, respectively, whereas the expression of BMP2 was 88% lower, in OA-affected cartilage than that of controls (all p < 0.03), but no difference was observed between hip and knee OA (all p > 0.4). The expression of TGFB1 was correlated with the expression of SMAD3 (rho = 0.50, p = 0.003) and MMP13 (rho = 0.46, p = 0.007) in OA-affected cartilage and the significance became stronger after adjustment for age, sex, and BMI. The expression of BMP2 was negatively correlated with both TGFB1 (rho = -0.50, p = 0.02) and MMP13 (rho = -0.48, p = 0.02) in healthy cartilage, but the significance was altered after adjustment for the covariates. There was no correlation between the expression of SMAD3 and MMP13.
  
  
  CONCLUSIONS
  Our results demonstrate that MMP13 expression is associated with an increased expression of TGFB1 in OA-affected cartilage, possibly through SMAD-independent TGF-β pathway. Furthermore, TGF-β/SMAD3 is overactivated in OA cartilage; yet, the consequence of this overactivation remains to be established.
authors.affiliation: Discipline of Genetics, Faculty of Medicine, Memorial University of Newfoundland, St. John's, NL, A1B 3V6, Canada. erfanae@mun.ca.
authors.forename: Erfan
authors.initials: E
authors.lastname: Aref-Eshghi
authors.affiliation: Discipline of Genetics, Faculty of Medicine, Memorial University of Newfoundland, St. John's, NL, A1B 3V6, Canada. ming.liu@med.mun.ca.
authors.forename: Ming
authors.initials: M
authors.lastname: Liu
authors.affiliation: Discipline of Genetics, Faculty of Medicine, Memorial University of Newfoundland, St. John's, NL, A1B 3V6, Canada. peh318@mun.ca.
authors.forename: Patricia E
authors.initials: PE
authors.lastname: Harper
authors.affiliation: Division of Biomedical Science, Faculty of Medicine, Memorial University of Newfoundland, St. John's, NL, A1B 3V6, Canada. jdore@mun.ca.
authors.forename: Jules
authors.initials: J
authors.lastname: Doré
authors.affiliation: Division of Orthopedics, Faculty of Medicine, Memorial University of Newfoundland, St. John's, NL, A1B 3V6, Canada. gmartin@munmed.ca.
authors.forename: Glynn
authors.initials: G
authors.lastname: Martin
authors.affiliation: Division of Orthopedics, Faculty of Medicine, Memorial University of Newfoundland, St. John's, NL, A1B 3V6, Canada. andrewfurey@hotmail.com.
authors.forename: Andrew
authors.initials: A
authors.lastname: Furey
authors.affiliation: Discipline of Genetics, Faculty of Medicine, Memorial University of Newfoundland, St. John's, NL, A1B 3V6, Canada. rcgreen@mun.ca.
authors.forename: Roger
authors.initials: R
authors.lastname: Green
authors.affiliation: Discipline of Medicine, Faculty of Medicine, Memorial University of Newfoundland, St. John's, NL, A1B 3V6, Canada. prahman@mun.ca.
authors.forename: Proton
authors.initials: P
authors.lastname: Rahman
authors.affiliation: Discipline of Genetics, Faculty of Medicine, Memorial University of Newfoundland, St. John's, NL, A1B 3V6, Canada. guangju.zhai@med.mun.ca.|Department of Twin Research & Genetic Epidemiology, King's College London, London, SE1 7EH, UK. guangju.zhai@med.mun.ca.
authors.forename: Guangju
authors.initials: G
authors.lastname: Zhai
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D055396:Bone Morphogenetic Protein 2 / Q000235:genetics; D002358:Cartilage, Articular / Q000378:metabolism* / Q000473:pathology / Q000601:surgery; D005260:Female; D015870:Gene Expression; D006801:Humans; D008297:Male; D053509:Matrix Metalloproteinase 13 / Q000235:genetics*; D008875:Middle Aged; D015207:Osteoarthritis, Hip / Q000235:genetics / Q000601:surgery; D020370:Osteoarthritis, Knee / Q000235:genetics / Q000601:surgery; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction / Q000235:genetics*; D051900:Smad3 Protein / Q000235:genetics*; D053773:Transforming Growth Factor beta1 / Q000235:genetics*
chemical_list: C525718:BMP2 protein, human; D055396:Bone Morphogenetic Protein 2; D051900:Smad3 Protein; C508960:TGFB1 protein, human; D053773:Transforming Growth Factor beta1; D053509:Matrix Metalloproteinase 13