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Ocu-miR-10a-5p promotes the chondrogenic differentiation of rabbit BMSCs by targeting BTRC-mediated Wnt/β-catenin signaling pathway

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Abstract

MicroRNAs (miRNAs) play an important role in articular cartilage damage in osteoarthritis (OA). However, the biological role of miRNAs in the chondrogenic differentiation of bone marrow mesenchymal stem cell (BMSC) remains largely unclear. Rabbit bone marrow mesenchymal stem cells (rBMSCs) were isolated, cultured, and identified. Afterwards, rBMSCs were induced to chondrogenic differentiation, examined by Alcian Blue staining. Differentially expressed miRNAs were identified in rBMSCs between induced and non-induced groups by miRNA sequencing analysis, part of which was validated via PCR assay. Cell viability and apoptosis were assessed by CCK-8 assay and Hoechst staining. Saffron O staining was utilized to assess chondrocyte hyperplasia. The expression of specific chondrogenic markers, including COL2A1, SOX9, Runx2, MMP-13, Aggrecan, and BMP-2, were measured at mRNA and protein levels. The association between beta-transducin repeat containing E3 ubiquitin protein ligase (BTRC) and miR-10a-5p in the miRNA family from rabbit (ocu-miR-10a-5p) was determined by luciferase reporter assay. A total of 76 differentially expressed miRNAs, including 52 downregulated and 24 upregulated miRNAs, were identified in rBMSCs from the induced group. Inhibition of ocu-miR-10a-5p suppressed rBMSC viability and chondrogenic differentiation, as well as downregulated the expression of β-catenin, SOX9, COL2A1, MMP-13, and Runx2. BTRC was predicted and confirmed as a target of ocu-miR-10a-5p. Overexpression of BTRC rescued the promoting impacts of overexpressed ocu-miR-10a-5p on chondrogenic differentiation of rBMSCs and β-catenin expression. Taken together, our data suggested that ocu-miR-10a-5p facilitated rabbit BMSC survival and chondrogenic differentiation by activating Wnt/β-catenin signaling through BTRC.

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Data availability

All data are available from the corresponding author with reasonable request.

References

  • Bartel DP (2004) MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 116:281–297

    Article  CAS  PubMed  Google Scholar 

  • Benary U, Kofahl B, Hecht A, Wolf J (2015) Mathematical modelling suggests a differential impact of β-transducin repeat-containing protein paralogues on Wnt/β-catenin signalling dynamics. FEBS J 282:1080–1096

    Article  CAS  PubMed  Google Scholar 

  • Cawthorn WP, Bree AJ, Yao Y, Du B, Hemati N, Martinez-Santibañez G, MacDougald OA (2012) Wnt6, Wnt10a and Wnt10b inhibit adipogenesis and stimulate osteoblastogenesis through a β-catenin-dependent mechanism. Bone 50:477–489

    Article  CAS  PubMed  Google Scholar 

  • Chen CZ, Li L, Lodish HF, Bartel DP (2004) MicroRNAs modulate hematopoietic lineage differentiation. Science 303:83–86

    Article  ADS  CAS  PubMed  Google Scholar 

  • Chen L, Liu G, Li W, Wu X (2019) Sonic hedgehog promotes chondrogenesis of rabbit bone marrow stem cells in a rotary cell culture system. BMC Dev Biol 19:18

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Cheng AM, Byrom MW, Shelton J, Ford LP (2005) Antisense inhibition of human miRNAs and indications for an involvement of miRNA in cell growth and apoptosis. Nucleic Acids Res 33:1290–1297

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Feng L, Yang Z, Li Y, Pan Q, Zhang X, Wu X, Lo JHT, Wang H, Bai S, Lu X, Wang M, Lin S, Pan X, Li G (2022) MicroRNA-378 contributes to osteoarthritis by regulating chondrocyte autophagy and bone marrow mesenchymal stem cell chondrogenesis. Mol Ther Nucleic Acids 28:328–341

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Goldring MB, Marcu KB (2012) Epigenomic and microRNA-mediated regulation in cartilage development, homeostasis, and osteoarthritis. Trends Mol Med 18:109–118

    Article  CAS  PubMed  Google Scholar 

  • He L, He T, Xing J, Zhou Q, Fan L, Liu C, Chen Y, Wu D, Tian Z, Liu B, Rong L (2020) Bone marrow mesenchymal stem cell-derived exosomes protect cartilage damage and relieve knee osteoarthritis pain in a rat model of osteoarthritis. Stem Cell Res Ther 11:276

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Hermann W, Lambova S, Muller-Ladner U (2018) Current treatment options for osteoarthritis. Curr Rheumatol Rev 14:108–116

    Article  CAS  PubMed  Google Scholar 

  • Hu N, Gao Y, Jayasuriya CT, Liu W, Du H, Ding J, Feng M, Chen Q (2019) Chondrogenic induction of human osteoarthritic cartilage-derived mesenchymal stem cells activates mineralization and hypertrophic and osteogenic gene expression through a mechanomiR. Arthritis Res Ther 21:167

    Article  PubMed  PubMed Central  Google Scholar 

  • Hu X, Shen N, Liu A, Wang W, Zhang L, Sui Z, Tang Q, Du X, Yang N, Ying W, Qin B, Li Z, Li L, Wang N, Lin H (2022) Bone marrow mesenchymal stem cell-derived exosomal miR-34c-5p ameliorates RIF by inhibiting the core fucosylation of multiple proteins. Mol Ther 30:763–781

    Article  CAS  PubMed  Google Scholar 

  • Hussain N, Zhu W, Jiang C, Xu J, Geng M, Wu X, Hussain S, Wang B, Rajoka MSR, Li Y, Tian J, Meng L, Lu S (2018) Down-regulation of miR-10a-5p promotes proliferation and restricts apoptosis via targeting T-box transcription factor 5 in inflamed synoviocytes. Biosci Rep 38:BSR20180003

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Hussain N, Zhu W, Jiang C, Xu J, Wu X, Geng M, Hussain S, Cai Y, Xu K, Xu P, Han Y, Sun J, Meng L, Lu S (2018) Down-regulation of miR-10a-5p in synoviocytes contributes to TBX5-controlled joint inflammation. J Cell Mol Med 22:241–250

    Article  CAS  PubMed  Google Scholar 

  • Jiang X, Liu J, Liu Q, Lu Z, Zheng L, Zhao J, Zhang X (2018) Therapy for cartilage defects: functional ectopic cartilage constructed by cartilage-simulating collagen, chondroitin sulfate and hyaluronic acid (CCH) hybrid hydrogel with allogeneic chondrocytes. Biomater Sci 6:1616–1626

    Article  CAS  PubMed  Google Scholar 

  • Jin Z, Da W, Zhao Y, Wang T, Xu H, Shu B, Gao X, Shi Q, Ma Y, Zhang Y, Wang Y, Tang D (2022) Role of skeletal muscle satellite cells in the repair of osteoporotic fractures mediated by β-catenin. J Cachexia Sarcopenia Muscle 13:1403–1417

    Article  PubMed  PubMed Central  Google Scholar 

  • Li Z, Wang Y, Xiang S, Zheng Z, Bian Y, Feng B, Weng X (2020) Chondrocytes-derived exosomal miR-8485 regulated the Wnt/β-catenin pathways to promote chondrogenic differentiation of BMSCs. Biochem Biophys Res Commun 523:506–513

    Article  CAS  PubMed  Google Scholar 

  • Liu J, Tang G, Liu W, Zhou Y, Fan C, Zhang W (2022) MiR-20a-5p facilitates cartilage repair in osteoarthritis via suppressing mitogen-activated protein kinase kinase kinase 2. Bioengineered 13:13801–13814

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ma Y, Wu Y, Chen J, Huang K, Ji B, Chen Z, Wang Q, Ma J, Shen S, Zhang J (2019) miR-10a-5p promotes chondrocyte apoptosis in osteoarthritis by targeting HOXA1. Mol Ther Nucleic Acids 14:398–409

    Article  CAS  PubMed  Google Scholar 

  • Mahboudi H, Soleimani M, Hanaee-Ahvaz H, Ghanbarian H, Bandehpour M, Enderami SE, Kazemi B (2018) New approach for differentiation of bone marrow mesenchymal stem cells toward chondrocyte cells with overexpression of microRNA-140. ASAIO J 64:662–672

    Article  CAS  PubMed  Google Scholar 

  • Marikawa Y, Elinson RP (1998) beta-TrCP is a negative regulator of Wnt/beta-catenin signaling pathway and dorsal axis formation in Xenopus embryos. Mech Dev 77:75–80

    Article  CAS  PubMed  Google Scholar 

  • Mayer U, Benditz A, Grassel S (2017) miR-29b regulates expression of collagens I and III in chondrogenically differentiating BMSC in an osteoarthritic environment. Sci Rep 7:13297

    Article  ADS  PubMed  PubMed Central  Google Scholar 

  • Mobasheri A, Kalamegam G, Musumeci G, Batt ME (2014) Chondrocyte and mesenchymal stem cell-based therapies for cartilage repair in osteoarthritis and related orthopaedic conditions. Maturitas 78:188–198

    Article  CAS  PubMed  Google Scholar 

  • Nie P, Li Y, Suo H, Jiang N, Yu D, Fang B (2019) Dasatinib promotes chondrogenic differentiation of human mesenchymal stem cells via the Src/Hippo-YAP signaling pathway. ACS Biomater Sci Eng 5:5255–5265

    Article  CAS  PubMed  Google Scholar 

  • Oichi T, Otsuru S, Usami Y, Enomoto-Iwamoto M, Iwamoto M (2020) Wnt signaling in chondroprogenitors during long bone development and growth. Bone 137:115368

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Schmidt I (2015) Surgical treatment options in thumb carpometacarpal osteoarthritis: a recent literature overview searching for practice pattern with special focus on total joint replacement. Curr Rheumatol Rev 11:39–46

    Article  PubMed  Google Scholar 

  • Shimada Y, Matsubayashi J, Saito A, Ohira T, Kuroda M, Ikeda N (2021) Small RNA sequencing to differentiate lung squamous cell carcinomas from metastatic lung tumors from head and neck cancers. PLoS ONE 16:e0248206

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Swingler TE, Wheeler G, Carmont V, Elliott HR, Barter MJ, Abu-Elmagd M, Donell ST, Boot-Handford RP, Hajihosseini MK, Munsterberg A, Dalmay T, Young DA, Clark IM (2012) The expression and function of microRNAs in chondrogenesis and osteoarthritis. Arthritis Rheum 64:1909–1919

    Article  CAS  PubMed  Google Scholar 

  • Wang JY, Yin CC, Wu CC, Geng SG, Yin M (2016) Icaritin promotes chondrogenic differentiation of BMSCs by Wnt/β-catenin signaling pathway. Zhongguo Zhong Yao Za Zhi 41:694–699

    CAS  PubMed  Google Scholar 

  • Wang L, Wang Q, Zhang XM (2014) Progress on bone marrow mesenchymal stem cells transplantation for spinal cord injury. Zhongguo Gu Shang 27:437–440

    CAS  PubMed  Google Scholar 

  • Xu S, Wu X (2019) miR-134 inhibits chondrogenic differentiation of bone marrow mesenchymal stem cells by targetting SMAD6. Biosci Rep 39:BSR20180921

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Xu Y, Jiang Y, Jia B, Wang Y, Li T (2021) Icariin stimulates osteogenesis and suppresses adipogenesis of human bone mesenchymal stem cells via miR-23a-mediated activation of the Wnt/β-catenin signaling pathway. Phytomedicine 85:153485

    Article  CAS  PubMed  Google Scholar 

  • Yu D, Li Z, Cao J, Shen F, Wei G (2022) microRNA-25-3p suppresses osteogenic differentiation of BMSCs in patients with osteoporosis by targeting ITGB3. Acta Histochem 124:151926

    Article  CAS  PubMed  Google Scholar 

  • Zhang P, Gao G, Zhou Z, He X (2021) microRNA-130b downregulation potentiates chondrogenic differentiation of bone marrow mesenchymal stem cells by targeting SOX9. Braz J Med Biol Res 54:e10345

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zhang Y, Le X, Zheng S, Zhang K, He J, Liu M, Tu C, Rao W, Du H, Ouyang Y, Li C, Wu D (2022) MicroRNA-146a-5p-modified human umbilical cord mesenchymal stem cells enhance protection against diabetic nephropathy in rats through facilitating M2 macrophage polarization. Stem Cell Res Ther 13:171

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zhang Y, Zhou L, Zhang Z, Ren F, Chen L, Lan Z (2020) miR-10a-5p inhibits osteogenic differentiation of bone marrow-derived mesenchymal stem cells. Mol Med Rep 22:135–144

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Funding

This study was supported by the Guangzhou Huadu District Science and Technology Plan Project (22-HDWS-069) and Traditional Chinese Medicine Bureau of Guangdong Province (No. 20211312).

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Author notes

  1. Donghua Liu and Wang Tang contributed equally to this article.

Authors and Affiliations

  1. Department of Spine Surgery, Guangzhou Hospital of Integrated Traditional and Western Medicine, Guangzhou, China

    Donghua Liu & Wang Tang

  2. Department of Joint Surgery, Guangzhou Hospital of Integrated Traditional and Western Medicine, 87 Yingbin Road, Huadu District, Guangzhou City, Guangdong Province, China

    Dongming Tang & Feng Jiao

  3. Guangzhou University of Chinese Medicine, Guangzhou, China

    Haixia Yan

Authors
  1. Donghua Liu
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  2. Wang Tang
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  3. Dongming Tang
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  4. Haixia Yan
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  5. Feng Jiao
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Contributions

(I) Conception and design: Donghua Liu, Wang Tang; (II) Administrative support: Feng Jiao; (III) Provision of study materials or patients: Dongming Tang; (IV) Collection and assembly of data: Haixia Yan; (V) Data analysis and interpretation: Donghua Liu, Wang Tang, Dongming Tang; (VI) Manuscript writing: All authors; (VII) Final approval of manuscript: All authors.

Corresponding author

Correspondence to Feng Jiao.

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Ethics statement

All the study participants signed an informed consent for the inclusion in the study. The study was approved by the ethics committee of Guangzhou Hospital of Integrated Traditional and Western Medicine. (No. E20210427).

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The authors declare no competing interests.

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Liu, D., Tang, W., Tang, D. et al. Ocu-miR-10a-5p promotes the chondrogenic differentiation of rabbit BMSCs by targeting BTRC-mediated Wnt/β-catenin signaling pathway. In Vitro Cell.Dev.Biol.-Animal (2024). https://doi.org/10.1007/s11626-024-00888-1

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  • DOI: https://doi.org/10.1007/s11626-024-00888-1

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