C/EBPβ and RUNX2 cooperate to degrade cartilage with MMP-13 as the target and HIF-2α as the inducer in chondrocytes

• Published in: Human molecular genetics 2012-03, Vol.21 (5), p.1111-1123
• Main Authors: Hirata, Makoto, Kugimiya, Fumitaka, Fukai, Atsushi, Saito, Taku, Yano, Fumiko, Ikeda, Toshiyuki, Mabuchi, Akihiko, Sapkota, Bishwa Raj, Akune, Toru, Nishida, Nao, Yoshimura, Noriko, Nakagawa, Takumi, Tokunaga, Katsushi, Nakamura, Kozo, Chung, Ung-il, Kawaguchi, Hiroshi
• Format: Article
• Language: English
• Subjects: Aged; Aged, 80 and over; Animals; Basic Helix-Loop-Helix Transcription Factors - metabolism; Biological and medical sciences; Bone Development; Cartilage - metabolism; CCAAT-Enhancer-Binding Protein-beta - genetics; CCAAT-Enhancer-Binding Protein-beta - metabolism; Cell Line, Tumor; Cells, Cultured; Chondrocytes - metabolism; Core Binding Factor Alpha 1 Subunit - genetics; Core Binding Factor Alpha 1 Subunit - metabolism; Fundamental and applied biological sciences. Psychology; Genetics of eukaryotes. Biological and molecular evolution; Humans; Matrix Metalloproteinase 13 - genetics; Matrix Metalloproteinase 13 - metabolism; Mice; Middle Aged; Molecular and cellular biology; Osteoarthritis - genetics; Osteoarthritis - metabolism; Osteoarthritis, Knee - genetics; Promoter Regions, Genetic; Transcription, Genetic; Transcriptional Activation
• ISSN: 0964-6906; 1460-2083
• DOI: 10.1093/hmg/ddr540

To elucidate the molecular mechanism underlying the endochondral ossification process during the skeletal growth and osteoarthritis (OA) development, we examined the signal network around CCAAT/enhancer-binding protein-β (C/EBPβ, encoded by CEBPB), a potent regulator of this process. Computational predictions and a C/EBP motif-reporter assay identified RUNX2 as the most potent transcriptional partner of C/EBPβ in chondrocytes. C/EBPβ and RUNX2 were induced and co-localized in highly differentiated chondrocytes during the skeletal growth and OA development of mice and humans. The compound knockout of Cebpb and Runx2 in mice caused growth retardation and resistance to OA with decreases in cartilage degradation and matrix metalloproteinase-13 (Mmp-13) expression. C/EBPβ and RUNX2 cooperatively enhanced promoter activity of MMP13 through specific binding to a C/EBP-binding motif and an osteoblast-specific cis-acting element 2 motif as a protein complex. Human genetic studies failed to show the association of human CEBPB gene polymorphisms with knee OA, nor was there a genetic variation around the identified responsive region in the human MMP13 promoter. However, hypoxia-inducible factor-2α (HIF-2α), a functional and genetic regulator of knee OA through promoting endochondral ossification, was identified as a potent and functional inducer of C/EBPβ expression in chondrocytes by the CEBPB promoter assay. Hence, C/EBPβ and RUNX2, with MMP-13 as the target and HIF-2α as the inducer, control cartilage degradation. This molecular network in chondrocytes may represent a therapeutic target for OA.