In vitro and in silico analysis of an inhibitory mechanism of osteoclastogenesis by salubrinal and guanabenz

Inactivating bone-resorbing osteoclasts is a prime therapeutic strategy for the prevention of bone loss in patients with osteopenia and osteoporosis. Synthetic agents such as salubrinal and guanabenz, which attenuate stress to the endoplasmic reticulum, are reported to inhibit development of osteocl...

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Bibliographic Details
Published in:Cellular signalling 2015-02, Vol.27 (2), p.353-362
Main Authors: Hamamura, Kazunori, Chen, Andy, Tanjung, Nancy, Takigawa, Shinya, Sudo, Akihiro, Yokota, Hiroki
Format: Article
Language:English
Subjects:
Acid Phosphatase - genetics
Acid Phosphatase - metabolism
Animals
Biocompatibility
Biomedical materials
Bone Marrow Cells - cytology
Bone Marrow Cells - drug effects
Bone Marrow Cells - metabolism
Bones
c-Fos
Cell Differentiation - drug effects
Cells, Cultured
Cellular
Cinnamates - pharmacology
Down-Regulation - drug effects
Gene expression
Guanabenz
Guanabenz - pharmacology
In vitro testing
Isoenzymes - genetics
Isoenzymes - metabolism
JunB
Mice
NFATC Transcription Factors - metabolism
NFATc1
Osteoblasts - cytology
Osteoblasts - drug effects
Osteoblasts - metabolism
Osteoclasts
Osteogenesis - drug effects
Patients
Principal Component Analysis
Proto-Oncogene Proteins c-fos - antagonists & inhibitors
Proto-Oncogene Proteins c-fos - genetics
Proto-Oncogene Proteins c-fos - metabolism
RANK Ligand - pharmacology
RNA Interference
RNA, Messenger - metabolism
RNA, Small Interfering - metabolism
Salubrinal
Tartrate-Resistant Acid Phosphatase
Thiourea - analogs & derivatives
Thiourea - pharmacology
Transcription Factor AP-1 - metabolism
Transcription Factors - antagonists & inhibitors
Transcription Factors - genetics
Transcription Factors - metabolism
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Summary:Inactivating bone-resorbing osteoclasts is a prime therapeutic strategy for the prevention of bone loss in patients with osteopenia and osteoporosis. Synthetic agents such as salubrinal and guanabenz, which attenuate stress to the endoplasmic reticulum, are reported to inhibit development of osteoclasts. However, the mechanism of their inhibitory action on osteoclasts is largely unknown. Using genome-wide expression profiles, we predicted key transcription factors that downregulated nuclear factor of activated T-cells, cytoplasmic 1 (NFATc1), a master transcription factor for osteoclastogenesis. Principal component analysis (PCA) predicted a list of transcription factors that were potentially responsible for reversing receptor activator of nuclear factor kappa-B ligand (RANKL)-driven stimulation of osteoclastogenesis. A partial silencing of NFATc1 allowed a selection of transcription factors that were likely to be located upstream of NFATc1. We validated the predicted transcription factors by focusing on two AP-1 transcription factors (c-Fos and JunB) using RAW264.7 pre-osteoclasts as well as primary bone marrow cells. As predicted, their mRNA and protein levels were elevated by RANKL, and the elevation was suppressed by salubrinal and guanabenz. A partial silencing of c-Fos or JunB by RNA interference decreased NFATc1 as well as tartrate-resistant acid phosphatase (TRAP) mRNA. Collectively, a systems-biology approach allows the prediction of a RANKL-salubrinal/guanabenz-NFATc1 regulatory axis, and in vitro assays validate an involvement of AP-1 transcription factors in suppression of osteoclastogenesis. •Salubrinal and guanabenz attenuate RANKL-driven osteoclastogenesis.•In silico analysis predicts genes involved in the RANKL–NFATc1 regulatory axis.•In vitro assays with RNA interference validate the role of c-Fos and JunB.
ISSN:0898-6568
1873-3913
DOI:10.1016/j.cellsig.2014.11.020