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miR-92a-3p-inspired shRNA exhibits pro-chondrogenic and chondrocyte protective effects in osteoarthritis treatment through targeting SMAD6/7

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Journal of Bone and Mineral Metabolism Aims and scope Submit manuscript

Abstract

Introduction

Osteoarthritis (OA) compromises patients’ quality of life and requires further study. Although miR-92a-3p was reported to possess chondroprotective effects, the underlying mechanism requires further clarification. The objectives of this study were to elucidate the mechanism by which miR-92a-3p alleviates OA and to examine the efficacy of shRNA-92a-3p, which was designed based on mature miR-92a-3p.

Materials and methods

TargetScan and luciferase reporter assay were used to predict the target of miR-92a-3p. Adipose-derived stem cells (ADSCs) were transfected with miR-92a-3p/miR-NC mimic for the analysis of chondrogenic biomarkers and SMAD proteins. ADSCs and osteoarthritic chondrocytes were transduced with shRNA-92a-3p for the analysis of chondrogenic biomarkers and SMAD proteins. OA was surgically induced in C57BL/6JJcl mice, and ADSCs with/without shRNA-92a-3p transduction were intra-articularly injected for the assessment of cartilage damage.

Results

SMAD6 and SMAD7 were predicted as direct targets of miR-92a-3p by TargetScan and luciferase reporter assay. Transfection of the miR-92a-3p mimic resulted in a decrease in SMAD6 and SMAD7 levels and an increase in phospho-SMAD2/3, phospho-SMAD1/5/9, SOX9, collagen type II, and aggrecan levels in ADSCs. Furthermore, shRNA-92a-3p decreased SMAD6 and SMAD7 levels, and increased phospho-SMAD2/3, phospho-SMAD1/5/9, SOX9, collagen type II, and aggrecan levels in ADSCs and osteoarthritic chondrocytes. Additionally, ADSC-shRNA-92a-3p-EVs reduced the rate of decrease of SOX9, collagen type II, and aggrecan in osteoarthritic chondrocytes. In mice with surgically induced OA, shRNA-92a-3p-treated ADSCs alleviated cartilage damage more effectively than nontreated ADSCs.

Conclusions

miR-92a-3p and shRNA-92a-3p exhibit therapeutic effects in treating OA by targeting SMAD6 and SMAD7, thereby enhancing TGF-β signaling.

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Acknowledgements

This work is supported by Japan Society for the Promotion of Science KAKENHI [grant numbers #19K22700, #21H03136]. P0 Human ADSCs were kindly provided by a previous laboratory member Dr. Inaki (Miyagi National Hospital). P0 mouse ADSCs were kindly provided by Dr. IGARASHI (the department of Sensory and Motor System Medicine, the University of Tokyo). P0 Human OA chondrocytes were kindly provided by Dr. Saito (the department of orthopedic surgery and spinal surgery, the University of Tokyo Hospital).

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Authors and Affiliations

  1. Department of Sensory and Motor System Medicine, Graduate School of Medicine, The University of Tokyo, Tokyo, 113-8655, Japan

    Chenhuang Zheng & Kazuto Hoshi

  2. Department of Tissue Engineering, The University of Tokyo Hospital, Bunkyo City, 7 Chome-3-1 Hongo, Tokyo, 113-8655, Japan

    Chenhuang Zheng, Kazuto Hoshi & Atsuhiko Hikita

  3. Department of Oral-Maxillofacial Surgery, Dentistry and Orthodontics, The University of Tokyo Hospital, Tokyo, 113-8655, Japan

    Kazuto Hoshi & Atsuhiko Hikita

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  1. Chenhuang Zheng
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Contributions

All authors contributed to the study conception and design. CZ prepared materials, performed the majority of the experiments described in this manuscript, and participated in the analysis of the histological data. AH and KH supervised the studies, provided technical support, and directed the interpretations of the result. Also, the first draft of the manuscript was written by CZ and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Atsuhiko Hikita.

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Conflict of interest

Atsuhiko Hikita held an endowed chair supported by FUJISOFT INCORPORATED (until 31 October 2020) and an endowed chair supported by CPC corporation, Kyowa Co., Ltd., Kanto Chemical Co., Inc., and Nichirei Corporation (from 1 July, 2021, to 30 June, 2022), and is affiliated with the social cooperation program of Kohjin Bio Co., Ltd. (since 1 July, 2022). All other authors have no conflicts of interest.

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Zheng, C., Hoshi, K. & Hikita, A. miR-92a-3p-inspired shRNA exhibits pro-chondrogenic and chondrocyte protective effects in osteoarthritis treatment through targeting SMAD6/7. J Bone Miner Metab 42, 1–16 (2024). https://doi.org/10.1007/s00774-023-01474-3

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  • DOI: https://doi.org/10.1007/s00774-023-01474-3

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