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Long noncoding RNA ZBTB40-IT1 regulates bone mass by directing the differentiation of human bone marrow mesenchymal stromal cells via the microRNA-514a-3p/FOXO4 axis

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Abstract

This study intended to clarify the mechanism of long noncoding RNA ZBTB40-IT1 in directing human bone marrow-derived mesenchymal stromal cell (hBMSC) differentiation. hBMSCs underwent osteogenic and adipogenic induction, and an osteoporosis mouse model was established via ovariectomy (OVX). Gain- and loss-of-function approaches were utilized in hBMSCs and mice to investigate the function of ZBTB40-IT1, microRNA (miR)-514a-3p, and forkhead box O4 (FOXO4). Dual-luciferase reporter and RNA pulldown assays were applied to evaluate the binding of miR-514a-3p to ZBTB40-IT1 or FOXO4. The femur of the OVX mice had upregulated ZBTB40-IT1 and FOXO4 expression and downregulated miR-514a-3p expression. The bone mass was increased in OVX mice through ZBTB40-IT1 or FOXO4 knockdown. ZBTB40-IT1 and FOXO4 were downregulated, whereas miR-514a-3p was upregulated in osteogenesis-induced hBMSCs, which was the opposite in adipogenesis-induced hBMSCs. ZBTB40-IT1 or FOXO4 knockdown or miR-514a-3p overexpression increased ARS/ALP absorbance and RUNX2 and OCN levels but decreased fat density and PPARγ and FABP4 levels in hBMSCs. Mechanistically, ZBTB40-IT1 elevated FOXO4 expression by binding to miR-514a-3p. miR-514a-3p inhibition annulled the effects of ZBTB40-IT1 downregulation on hBMSC osteogenesis and adipogenesis, and FOXO4 overexpression abolished the impacts of miR-514a-3p upregulation on hBMSC osteogenesis and adipogenesis. Conclusively, ZBTB40-IT1 inhibition promotes the osteogenic differentiation of hBMSCs via the miR-514a-3p/FOXO4 axis, thereby increasing bone mass.

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Acknowledgements

Thanks for all the contributors and participants.

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Authors and Affiliations

  1. Department of Orthopedics, Nanfang Hospital, Southern Medical University, No. 1838, North Guangzhou Street, Baiyun District, Guangzhou, 510515, Guangdong, China

    Zhe Shi, Qiang Zhong & Yuhang Chen

  2. Rehabilitation Medical School, Guangzhou International Economics College, Guangzhou, 510540, Guangdong, China

    Xin Luo

Authors
  1. Zhe Shi
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  2. Qiang Zhong
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  3. Yuhang Chen
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  4. Xin Luo
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Contributions

ZS: conceived the ideas. ZS: designed the experiments. QZ: performed the experiments. QZ: analyzed the data. YC and XL: provided critical materials. YC and XL: wrote the manuscript. ZS: supervised the study. All the authors have read and approved the final version for publication.

Corresponding author

Correspondence to Zhe Shi.

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The authors report no relationships that could be construed as a conflict of interest.

Ethical approval

All patients signed an informed consent form, and the experimental design was approved by the Ethics Committee of Nanfang Hospital, Southern Medical University (approval number: NFYY-2019-0541). All animal care, treatment, and surgical techniques followed the protocols approved by the Animal Care and Use Committee of Nanfang Hospital, Southern Medical University. Mice were used for this study under the “Implementation Rules for the Management of Medical Laboratory Animals” promulgated by the Chinese Health and Planning Commission and the “Guidelines for the Use and Management of Laboratory Animals” by the National Institutes of Health.

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Shi, Z., Zhong, Q., Chen, Y. et al. Long noncoding RNA ZBTB40-IT1 regulates bone mass by directing the differentiation of human bone marrow mesenchymal stromal cells via the microRNA-514a-3p/FOXO4 axis. Human Cell 35, 1408–1423 (2022). https://doi.org/10.1007/s13577-022-00730-4

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  • DOI: https://doi.org/10.1007/s13577-022-00730-4

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