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miR-764-5p promotes osteoblast differentiation through inhibition of CHIP/STUB1 expression
Junwei Guo1, Fangli Ren, Yinyin Wang
1State Key Laboratory of Biomembrane and Membrane Biotechnology, School of Medicine, Tsinghua University, Beijing, China.
Abstract:
Differentiation of committed precursor cells into the osteoblast lineage is tightly regulated by several factors, including Runx2 and BMP2. We previously reported that C terminus of Hsc70-interacting protein/STIP1 homology and U-Box containing protein 1 (CHIP/STUB1) negatively regulated osteoblast differentiation through promoting Runx2 protein degradation. However, how CHIP is regulated during osteoblast differentiation remains unknown. In this study, we found that miR-764-5p is up-expressed during the osteoblast differentiation in calvarial and osteoblast progenitor cells, coupled with down-expression of CHIP protein. We observed that forced expression or inhibition of miR-764-5p decreased or increased the CHIP protein level through affecting its translation by targeting the 3'-UTR region. Perturbation of miR-764-5p resulted in altered differentiation fate of osteoblast progenitor cells and the role of miR-764-5p was reversed by overexpression of CHIP, whereas depletion of CHIP impaired the effect of miR-764-5p. Our data showed that miR-764-5p positively regulates osteoblast differentiation from osteoblast progenitor cells by repressing the translation of CHIP protein.
Insights
MicroRNA-764-5p promotes osteoblast differentiation by reducing CHIP protein levels. This microRNA targets CHIP mRNA, inhibiting its translation and positively impacting bone cell development.
Area of Science:
- Molecular Biology
- Cell Biology
- Biochemistry
Background:
- Osteoblast differentiation is crucial for bone formation and is regulated by key factors like Runx2 and BMP2.
- CHIP (C-terminus of Hsc70-interacting protein/STIP1 homology and U-box containing protein 1) negatively regulates osteoblast differentiation by degrading Runx2.
- The precise regulation of CHIP during osteoblast differentiation is currently unknown.
Purpose of the Study:
- To investigate the role and regulation of CHIP during osteoblast differentiation.
- To identify microRNAs involved in modulating CHIP expression in osteoblasts.
- To elucidate the functional impact of miR-764-5p on osteoblast progenitor cell differentiation.
Main Methods:
- Quantitative real-time PCR to measure miR-764-5p and CHIP expression.
- Western blotting to assess CHIP protein levels.
- Luciferase reporter assays to confirm targeting of the CHIP 3'-UTR by miR-764-5p.
- Gain- and loss-of-function experiments for miR-764-5p and CHIP in osteoblast progenitor cells.
Main Results:
- miR-764-5p expression is upregulated during osteoblast differentiation in calvarial and progenitor cells, inversely correlating with CHIP protein levels.
- miR-764-5p directly targets the 3'-UTR of CHIP mRNA, inhibiting its translation.
- Modulating miR-764-5p levels altered osteoblast progenitor cell differentiation, with effects reversed by CHIP manipulation.
Conclusions:
- miR-764-5p positively regulates osteoblast differentiation from progenitor cells.
- This regulation occurs through the repression of CHIP protein translation.
- miR-764-5p represents a novel regulator in the osteoblast differentiation pathway.
