Bromodomain and Extra-terminal Proteins (BET) Inhibitors Suppress Chondrocyte Differentiation and Restrain Bone Growth.

Small-molecule inhibitors for bromodomain and extra-terminal proteins (BET) have recently emerged as potential therapeutic agents in clinical trials for various cancers. However, to date, it is unknown whether these inhibitors have side effects on bone structures. Here, we report that inhibition of BET bromodomain proteins may suppress chondrocyte differentiation and restrain bone growth. We generated a luciferase reporter system using the chondrogenic cell line, ATDC5, in which the luciferase gene was driven by the promoter of Col2a1, an elementary collagen of chondrocyte. The COL2A1-luc ATDC5 system was used for rapidly screening both activators and repressors of human collagen COL2A1 gene expression, and we found that BET bromodomain inhibitors reduce the COL2A1-luc. Consistent with the luciferase assay, BET inhibitors decrease the expression of Col2a1. Furthermore, we constructed a zebrafish line in which the Enhanced Green Fluorescent Protein (EGFP) expression was driven by Col2a1 promoter. The transgenic (Col2a1-EGFP) zebrafish line demonstrated that BET inhibitors I-BET151 and (+)-JQ1 may affect EGFP expression in zebrafish. Furthermore, we found that I-BET151 and (+)-JQ1 may affect chondrocyte differentiation in vitro and inhibit zebrafish growth in vivo. Mechanistic analysis revealed that BET inhibitors influenced the depletion of RNA polymerase II from the Col2a1 promoter. Collectively, these results suggest that BET bromodomain inhibition may have side effects on skeletal bone structures.