BAPX‐1/NKX‐3.2 Acts as a Chondrocyte Hypertrophy Molecular Switch in Osteoarthritis

• Published in: Arthritis & rheumatology (Hoboken, N.J.) N.J.), 2015-11, Vol.67 (11), p.2944-2956
• Main Authors: Caron, Marjolein M. J., Emans, Pieter J., Surtel, Don A. M., van der Kraan, Peter M., van Rhijn, Lodewijk W., Welting, Tim J. M.
• Format: Article
• Language: English
• Subjects: Animals; Arthritis, Experimental - metabolism; Arthritis, Experimental - pathology; Bone Morphogenetic Protein 7 - pharmacology; Cartilage, Articular - drug effects; Cartilage, Articular - metabolism; Cartilage, Articular - pathology; Cell Differentiation; Cell Enlargement - drug effects; Chondrocytes - drug effects; Chondrocytes - metabolism; Chondrocytes - pathology; Gene expression; Genotype & phenotype; Homeodomain Proteins - metabolism; Humans; Interleukin-1beta - pharmacology; Mice; Osteoarthritis - metabolism; Osteoarthritis - pathology; Rodents; Transcription Factors - metabolism; Tumor Necrosis Factor-alpha - pharmacology
• ISSN: 2326-5191; 2326-5205
• DOI: 10.1002/art.39293

Objective Osteoarthritis (OA) development involves a shift of the articular chondrocyte phenotype toward hypertrophic differentiation via still poorly characterized mechanisms. The purpose of this study was to test our hypothesis that the function of BAPX‐1/NKX‐3.2 is impaired in OA chondrocytes and leads directly to loss of hypertrophic protection of the articular chondrocyte, which is central in the changing chondrocyte phenotype that drives OA. Methods Human articular chondrocytes (HACs; from healthy and OA donors) and SW‐1353 chondrocytic cells were exposed to bone morphogenetic protein 7 (BMP‐7), interleukin‐1β (IL‐1β), tumor necrosis factor, or OA synovial fluid (SF; 20% [volume/volume]). Loss‐of‐function and gain‐of‐function experiments for BAPX‐1/NKX‐3.2 were performed. Mouse experimental models of OA were used, and (immuno)histochemistry of tissue sections was performed. Gene and protein expression of BAPX‐1/NKX‐3.2 and chondrogenic, hypertrophic, and OA‐related mediators were determined by real‐time quantitative polymerase chain reaction analysis and immunoblotting. In addition, alkaline phosphatase (AP) activity and prostaglandin E2 levels were measured. Results BAPX‐1/NKX‐3.2 expression correlated negatively with expression of chondrocyte hypertrophic markers (RUNX‐2, COL10A1, AP), cartilage‐degrading enzymes (matrix metalloproteinase 13, ADAMTS‐5), and mediators of inflammation (cyclooxygenase 2, IL‐6) in healthy and OA chondrocytes, as well as in OA induced chondrocytes. BAPX‐1/NKX‐3.2 positivity was diminished in articular chondrocytes in the knee joints of mice with experimental OA. Knockdown of BAPX‐1/NKX‐3.2 in HACs did not influence the expression of SOX9, COL2A1, or aggrecan, but led to an acute hypertrophic shift in the HAC phenotype. Overexpression of BAPX‐1/NKX‐3.2 decreased hypertrophic gene expression in HACs. Furthermore, the hypertrophic OA chondrocyte phenotype could be counteracted by overexpression of BAPX‐1/NKX‐3.2 and by BMP‐7 in a BAPX‐1/NKX‐3.2 dependent manner. Conclusion Our findings indicate that BAPX‐1/NKX‐3.2 is a molecular switch that is involved in controlling the hypertrophic phenotype of the postdevelopmental (OA) articular chondrocyte.