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MEG3 alleviates ankylosing spondylitis by suppressing osteogenic differentiation of mesenchymal stem cells through regulating microRNA-125a-5p-mediated TNFAIP3

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Abstract

Osteoblasts are important regulators of bone formation, but their roles in ankylosing spondylitis (AS) remain unclear. This study aims to explore the role of long non-coding RNA (lncRNA) maternally expressed 3 (MEG3) MEG3 in AS. Serum from AS patients as well as AS mesenchymal stem cells (ASMSCs) and healthy donors mesenchymal stem cells (HDMSCs) was collected. Accordingly, poorly expressed MEG3 and TNF alpha induced protein 3 (TNFAIP3) as well as overexpressed microRNA-125a-5p (miR-125a-5p) were noted in the serum of AS patients and in ASMSCs during the osteogenic induction process. Meanwhile, the interaction among MEG3, miR-125a-5p, and TNFAIP3 was determined and their effect on osteoblast activity was examined in vitro and in vivo. Overexpression of MEG3 and TNFAIP3 or inhibition of miR-125a-5p was found to inactivate the Wnt/β-catenin pathway, thus suppressing osteogenic differentiation of MSCs. MEG3 competitively bound to miR-125a-5p to increase TNFAIP3 expression, thereby inactivating the Wnt/β-catenin pathway and repressing the osteogenic differentiation of MSCs. In proteoglycan (PG)-induced AS mouse models, MEG3 also reduced osteogenic activity of MSCs to inhibit AS progression through the miR-125a-5p/TNFAIP3/Wnt/β-catenin axis. Therefore, up-regulation of MEG3 or depletion of miR-125a-5p holds potential of alleviating AS, which sheds light on a new therapeutic strategy for AS treatment.

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Data availability

The datasets generated/analysed during the current study are available from the corresponding author on reasonable request.

Abbreviations

AS:

Ankylosing spondylitis

lncRNA:

Long non-coding RNA

MEG3:

Maternally expressed 3

ASMSCs:

AS mesenchymal stem cells

HDMSCs:

Healthy donors mesenchymal stem cells

miR-125a-5p:

microRNA-125a-5p

TNFAIP3:

TNF alpha induced protein 3

GM:

Growth medium

ALP:

Alkaline phosphatase

RIPA:

Radio-immunoprecipitation assay

CST:

Cell signaling technologies

IHC:

Immunohistochemistry

DAB:

Diaminobenzidine

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Acknowledgements

Not applicable.

Funding

This work was supported by the Youth Science Foundation of Guangxi Medical University, Grant/Award Number: GXMUYFY201712; Guangxi Young and Middle aged Teacher’s Basic Ability Promoting Project, Grant/Award Number: 2019KY0119.

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Authors and Affiliations

  1. Spine and Osteopathy Ward, The First Affiliated Hospital of Guangxi Medical University, No. 6, Shuangyong Road, Nanning, 530021, Guangxi Zhuang Autonomous Region, People’s Republic of China

    Chong Liu & Xinli Zhan

  2. Guangxi Medical University, Nanning, 530021, People’s Republic of China

    Tuo Liang, Zide Zhang, Jiarui Chen & Jang Xue

  3. Reproductive Medicine Center, The First Affiliated Hospital of Guangxi Medical University, No.6 Shuangyong Road, Nanning, 530021, Guangxi Zhuang Autonomous Region, People’s Republic of China

    Liang Ren

Authors
  1. Chong Liu
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  4. Jiarui Chen
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Contributions

CL and TL designed study and collated data, ZZ and JC carried out data analyses and produced initial draft of manuscript, JX, XZ and LR contributed to drafting manuscript. All authors read and approved final manuscript.

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Correspondence to Xinli Zhan or Liang Ren.

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Ethical approval

All research procedures were approved by the Ethics Committee of The First Affiliated Hospital of Guangxi Medical University and in line with the Declaration of Helsinki. All animal experiments were approved by the Animal Ethics Committee of The First Affiliated Hospital of Guangxi Medical University.

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Supplementary Information

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10495_2022_1804_MOESM1_ESM.jpg

Supplementary file1 (JPG 254 kb). Fig. S1 Chondrogenesis in pellet cultures. Cartilage beads formed by MSCs cultured under chondrogenic conditions for 1, 2 and 3 weeks with Alcian-blue staining

10495_2022_1804_MOESM2_ESM.jpg

Supplementary file2 (JPG 423 kb). Fig. S2 miR-125a-5p expression in ASMSCs and HDMSCs treated with LV- miR-125a-5p detected by RT-qPCR. ***p < 0.001. The measurement data were expressed as mean ± standard deviation, and data between two groups were compared with independent sample t test. The cell experiment was conducted in triplicates

10495_2022_1804_MOESM3_ESM.jpg

Supplementary file3 (JPG 8990 kb). Fig. S3 Differentially expressed mRNAs in AS samples screened by bioinformatics analyses. A, A heat map of the expression of the differentially expressed mRNAs in AS patient samples from the GSE11886 microarray. B, Volcano map of differentially expressed mRNAs in AS patient samples from the GSE11886 microarray. C, Veen diagram of the downstream target genes of miR-125a-5p predicted by starBase (http://starbase.sysu.edu.cn/index.php), TargetScan (http://www.targetscan.org/vert_71/), mirDIP (http://ophid.utoronto.ca/mirDIP/index.jsp#R) and miRDB (http://www.mirdb.org/) databases and the significantly down-regulated genes in the GSE11886 microarray. Blue circle indicates target genes predicted by starBase database, red circle indicates target genes predicted by TargetScan database, green circle indicates target genes predicted by mirDIP database, yellow circle indicates target genes predicted by miRDB database, orange circle indicates the significantly down-regulated genes in AS patient samples in GSE11886 microarray, and the middle part indicates the intersection of the target genes predicted by these five datasets

Supplementary file4 (DOCX 15 kb)

Supplementary file5 (DOCX 16 kb)

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Liu, C., Liang, T., Zhang, Z. et al. MEG3 alleviates ankylosing spondylitis by suppressing osteogenic differentiation of mesenchymal stem cells through regulating microRNA-125a-5p-mediated TNFAIP3. Apoptosis 28, 498–513 (2023). https://doi.org/10.1007/s10495-022-01804-2

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