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A potential therapeutic target for regulating osteoporosis via suppression of osteoclast differentiation
Qin Sun1, Boran Zhang1, Wei Zhu1
1Department of Stomatology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Objectives:
Osteoclast differentiation is regulated by transcriptional, post-transcriptional and post-translational mechanisms. Micro-ribonucleic acids (miRNAs) are 20-24 nucleotides long non-coding RNAs involved in post-translational regulation of gene expressions during osteoclast differentiation. The objective of the present study was to investigate the role played by the miRNA, miR-338-3p, in osteoclastogenesis.
Methods:
Osteoclastogenesis was induced in murine RAW264.7 cells using M-CSF and RANKL. The differentiated cells were harvested at designated times for TRAP staining and detection of designated gene expressions. A synthetic miR-338-3p mimic or its inhibitor was transfected into RAW264.7 cells prior to the induction of osteoclastogenesis. The effects of mimic or inhibitor on osteoclastogenesis were examined by qRT-PCR and TRAP staining. Bioinformatic analysis and luciferase activity were performed to identify the relationship between miR-338-3p and the transcription factor MafB. The miR-338-3p mimic and MafB siRNA were co-transfected into RAW264.7 cells to evaluate the cross-talk between miR-338-3p and MafB.
Results:
miR-338-3p was increased significantly during osteoclast differentiation. Overexpression of miR-338-3p promoted osteoclastogenesis while its inhibition had the opposite effect. Bioinformatic analysis and dual luciferase assays indicated that miR-338-3p targeted MafB to repress its gene expression. MafB knockdown by RNA silencing blocked the promotional effect of miR-338-3p on osteoclast differentiation.
Conclusion:
Because miR-338-3p is crucial for osteoclastic differentiation via targeting of the transcription factor MafB, inhibition of this miRNA represents a potential strategy for modulating osteoporosis in an aging population. CLINICAL SIGNIfiCANCE: Understanding the role played by miR-338-3p in osteoclast differentiation bridges the gap between the pathogenesis of osteoporosis and the quest for novel therapeutics to reduce the risk of bone fracture associated with this global disease.
Insights
MicroRNA miR-338-3p promotes osteoclast differentiation by targeting the transcription factor MafB. Inhibiting miR-338-3p offers a potential therapeutic strategy for osteoporosis, particularly in aging populations.
Area of Science:
- Molecular Biology
- Cell Biology
- Biochemistry
Background:
- Osteoclast differentiation is a complex process regulated by multiple mechanisms.
- Micro-ribonucleic acids (miRNAs) are key post-transcriptional regulators of gene expression during osteoclastogenesis.
Purpose of the Study:
- To investigate the specific role of microRNA miR-338-3p in the process of osteoclast differentiation (osteoclastogenesis).
Main Methods:
- Osteoclastogenesis was induced in RAW264.7 cells.
- miR-338-3p mimic and inhibitor transfections were performed.
- Gene expression was analyzed using qRT-PCR and TRAP staining.
- Bioinformatic analysis and luciferase assays identified the target gene MafB.
- MafB knockdown was used to assess cross-talk with miR-338-3p.
Main Results:
- miR-338-3p levels significantly increased during osteoclast differentiation.
- Overexpression of miR-338-3p enhanced osteoclastogenesis; inhibition reduced it.
- miR-338-3p directly targets and represses the transcription factor MafB.
- MafB knockdown abolished the pro-osteoclastogenic effect of miR-338-3p.
Conclusions:
- miR-338-3p plays a crucial role in osteoclast differentiation by targeting MafB.
- Inhibition of miR-338-3p presents a potential therapeutic avenue for osteoporosis.
- Understanding miR-338-3p's role aids in developing novel treatments for bone fractures.
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