title: Zoledronate inhibits receptor activator of nuclear factor kappa-B ligand-induced osteoclast differentiation via suppression of expression of nuclear factor of activated T-cell c1 and carbonic anhydrase 2.
abstract: Bisphosphonates (BPs) are widely used in the prevention of skeletal-related events (SRE), including osteoporosis, skeletal metastases of malignant tumours, and multiple myeloma. Osteonecrosis of the jaw (ONJ) is frequently reported as a major adverse effect induced by BP treatment. The receptor activator of the nuclear factor kappa-B ligand (RANKL) inhibitor, denosumab, has recently been used to prevent SRE, but the frequency of ONJ induced by denosumab is similar to that by BPs. This finding suggests that the inhibition of RANKL-mediated osteoclastogenesis may have a close relationship with the occurrence of ONJ. We therefore investigated the expression status of RANKL-inducible genes in zoledronate-treated mouse osteoclast precursor cells. The molecular targets of zoledronate in the RANKL signal pathway and additional factors associated with osteoclastogenesis were analysed by genome-wide screening. Microarray analysis identified that among 31 genes on 44 entities of RANKL-inducible genes, the mRNA expression level of two genes, i.e., nuclear factor of activated T-cells c1 (NFATc1) and carbonic anhydrase 2 (CAII), was decreased in zoledronate-treated cells. Subsequent analyses verified that these two genes were significantly silenced by zoledronate treatment and that their expression was restored following inhibition of zoledronate action by geranylgeraniol. Zoledronate inhibited RANKL-induced osteoclast differentiation by suppression of NFATc1 and CAII gene expression. Our results suggest that these genes might be common targets for zoledronate and denosumab in the mechanism underlying RANKL-induced osteoclast differentiation. A clear understanding of the common molecular mechanisms of bone-remodelling agents is thus essential for prevention of ONJ.
authors.affiliation: Department of Oral and Maxillofacial Surgery, Applied Life Sciences, Institute of Biomedical and Health Sciences, Hiroshima University, Kasumi 1-2-3, Minami-ku, Hiroshima 734-8553, Japan. Electronic address: tnakag@hiroshima-u.ac.jp.
authors.forename: Takayuki
authors.initials: T
authors.lastname: Nakagawa
authors.affiliation: Department of Oral and Maxillofacial Surgery, Applied Life Sciences, Institute of Biomedical and Health Sciences, Hiroshima University, Kasumi 1-2-3, Minami-ku, Hiroshima 734-8553, Japan.
authors.forename: Kouji
authors.initials: K
authors.lastname: Ohta
authors.affiliation: Department of Oral and Maxillofacial Surgery, Applied Life Sciences, Institute of Biomedical and Health Sciences, Hiroshima University, Kasumi 1-2-3, Minami-ku, Hiroshima 734-8553, Japan.
authors.forename: Kazumi
authors.initials: K
authors.lastname: Kubozono
authors.affiliation: Department of Oral and Maxillofacial Surgery, Applied Life Sciences, Institute of Biomedical and Health Sciences, Hiroshima University, Kasumi 1-2-3, Minami-ku, Hiroshima 734-8553, Japan.
authors.forename: Yoko
authors.initials: Y
authors.lastname: Ishida
authors.affiliation: Department of Oral and Maxillofacial Surgery, Applied Life Sciences, Institute of Biomedical and Health Sciences, Hiroshima University, Kasumi 1-2-3, Minami-ku, Hiroshima 734-8553, Japan.
authors.forename: Takako
authors.initials: T
authors.lastname: Naruse
authors.affiliation: Department of Oral and Maxillofacial Surgery, Applied Life Sciences, Institute of Biomedical and Health Sciences, Hiroshima University, Kasumi 1-2-3, Minami-ku, Hiroshima 734-8553, Japan.
authors.forename: Masaaki
authors.initials: M
authors.lastname: Takechi
authors.affiliation: Department of Oral and Maxillofacial Surgery, Applied Life Sciences, Institute of Biomedical and Health Sciences, Hiroshima University, Kasumi 1-2-3, Minami-ku, Hiroshima 734-8553, Japan.
authors.forename: Nobuyuki
authors.initials: N
authors.lastname: Kamata
mesh_terms: D000135:Acid Phosphatase / Q000378:metabolism; D000818:Animals; D002256:Carbonic Anhydrases / Q000235:genetics / Q000378:metabolism*; D002454:Cell Differentiation / Q000187:drug effects; D004164:Diphosphonates / Q000633:toxicity*; D004224:Diterpenes / Q000494:pharmacology; D015870:Gene Expression; D007093:Imidazoles / Q000633:toxicity*; D007527:Isoenzymes / Q000378:metabolism; D051379:Mice; D046228:Microarray Analysis; D050778:NFATC Transcription Factors / Q000235:genetics / Q000378:metabolism*; D010010:Osteoclasts / Q000187:drug effects*; D053245:RANK Ligand / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D015398:Signal Transduction / Q000187:drug effects; D000071681:Tartrate-Resistant Acid Phosphatase; D000077211:Zoledronic Acid
chemical_list: D004164:Diphosphonates; D004224:Diterpenes; D007093:Imidazoles; D007527:Isoenzymes; D050778:NFATC Transcription Factors; D053245:RANK Ligand; D012333:RNA, Messenger; D000077211:Zoledronic Acid; C040155:geranylgeranic acid; D000135:Acid Phosphatase; D000071681:Tartrate-Resistant Acid Phosphatase; D002256:Carbonic Anhydrases
keywords: Bisphosphonate; CAII; DNA microarray; NFATc1; RANKL; Zoledronate