C/ebpα controls osteoclast terminal differentiation, activation, function, and postnatal bone homeostasis through direct regulation of Nfatc1

Osteoclast lineage commitment and differentiation have been studied extensively, although the mechanism by which transcription factor(s) control osteoclast terminal differentiation, activation, and function remains unclear. CCAAT/enhancer-binding protein alpha (C/ebp alpha) has been reported to be a key regulator of osteoclast cell lineage commitment, yet C/ebp alpha's roles in osteoclast terminal differentiation, activation and function, and bone homeostasis, under physiological or pathological conditions, have not been studied because newborn C/ebp alpha-null mice die within several hours after birth. Furthermore, the function of C/ebp alpha in osteoclast terminal differentiation, activation, and function is largely unknown. Herein, we generated and analyzed an osteoclast-specific C/ebp alpha conditional knockout (CKO) mouse model via Ctsk-Cre mice and found that C/ebp alpha-deficient mice exhibited a severe osteopetrosis phenotype due to impaired osteoclast terminal differentiation, activation, and function, including mildly reduced osteoclast number, impaired osteoclast polarization, actin formation, and bone resorption, which demonstrated the novel function of C/ebp alpha in cell function and terminal differentiation. Interestingly, C/ebp alpha deficiency did not affect bone formation or monocyte/macrophage development. Our results further demonstrated that C/ebp alpha deficiency suppressed the expression of osteoclast functional genes, e.g. encoding cathepsin K (Ctsk), Atp6i (Tcirg1), and osteoclast regulator genes, e.g. encoding c-fos (Fos), and nuclear factor of activated T-cells 1 (Nfatc1), while having no effect on Pu.1 (Spi1) expression. Promoter activity mapping and ChIP assay defined the critical cis-regulatory element (CCRE) in the promoter region of Nfatc1, and also showed that the CCREs were directly associated with C/ebp alpha, which enhanced the promoter's activity. The deficiency of C/ebp alpha in osteoclasts completely blocked ovariectomy-induced bone loss, indicating that C/ebp alpha is a promising new target for the treatment of osteolytic diseases. Copyright (C) 2017 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.