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Transcription factor 21 accelerates vascular calcification in mice by activating the IL-6/STAT3 signaling pathway and the interplay between VSMCs and ECs

Acta Pharmacologica Sinica volume 44, pages 1625–1636 (2023)Cite this article

Abstract

Vascular calcification is caused by the deposition of calcium salts in the intimal or tunica media layer of the aorta, which increases the risk of cardiovascular events and all-cause mortality. However, the mechanisms underlying vascular calcification are not fully clarified. Recently it has been shown that transcription factor 21 (TCF21) is highly expressed in human and mouse atherosclerotic plaques. In this study we investigated the role of TCF21 in vascular calcification and the underlying mechanisms. In carotid artery atherosclerotic plaques collected from 6 patients, we found that TCF21 expression was upregulated in calcific areas. We further demonstrated TCF21 expression was increased in an in vitro vascular smooth muscle cell (VSMC) osteogenesis model. TCF21 overexpression promoted osteogenic differentiation of VSMC, whereas TCF21 knockdown in VSMC attenuated the calcification. Similar results were observed in ex vivo mouse thoracic aorta rings. Previous reports showed that TCF21 bound to myocardin (MYOCD) to inhibit the transcriptional activity of serum response factor (SRF)-MYOCD complex. We found that SRF overexpression significantly attenuated TCF21-induced VSMC and aortic ring calcification. Overexpression of SRF, but not MYOCD, reversed TCF21-inhibited expression of contractile genes SMA and SM22. More importantly, under high inorganic phosphate (3 mM) condition, SRF overexpression reduced TCF21-induced expression of calcification-related genes (BMP2 and RUNX2) as well as vascular calcification. Moreover, TCF21 overexpression enhanced IL-6 expression and downstream STAT3 activation to facilitate vascular calcification. Both LPS and STAT3 could induce TCF21 expression, suggesting that the inflammation and TCF21 might form a positive feedback loop to amplify the activation of IL-6/STAT3 signaling pathway. On the other hand, TCF21 induced production of inflammatory cytokines IL-1β and IL-6 in endothelial cells (ECs) to promote VSMC osteogenesis. In EC-specific TCF21 knockout (TCF21ECKO) mice, VD3 and nicotine-induced vascular calcification was significantly reduced. Our results suggest that TCF21 aggravates vascular calcification by activating IL-6/STAT3 signaling and interplay between VSMC and EC, which provides new insights into the pathogenesis of vascular calcification.

TCF21 enhances vascular calcification by activating the IL-6-STAT3 signaling pathway. TCF21 inhibition may be a new potential therapeutic strategy for the prevention and treatment of vascular calcification.

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Fig. 1: TCF21 was positively correlated with vascular calcification.
Fig. 2: TCF21 promotes HASMC osteogenesis.
Fig. 3: TCF21 enhances media calcification in the thoracic aorta ex vivo.
Fig. 4: SRF abolishes TCF21-induced calcification.
Fig. 5: TCF21 activates the IL-6-STAT3 signaling pathway.
Fig. 6: TCF21 promotes calcification through ECs and VSMCs interactions.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (NSFC) Grants U22A20272 to YLC and YJD, 82160094 to MXJ; Natural Science Foundation of Anhui province 2208085MH196 to BCZ.

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Authors and Affiliations

  1. Key Laboratory of Metabolism and Regulation for Major Diseases of Anhui Higher Education Institutes, School of Food and Biological Engineering, Hefei University of Technology, Hefei, 230009, China

    Xiao-kang Zhao, Meng-meng Zhu, Sheng-nan Wang, Ting-ting Zhang, Xiao-ning Wei, Cheng-yi Wang, Juan Zheng, Wen-ya Zhu, Xiao-xiao Yang, Ji-hong Han & Yuan-li Chen

  2. The Institute of Translational Medicine, the National Engineering Research Center for Bioengineering Drugs and the Technologies, Nanchang University, Nanchang, 330031, China

    Mei-xiu Jiang

  3. Department of Endocrinology, The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, 230036, China

    Suo-wen Xu

  4. School of Pharmacy, Bengbu Medical College, Bengbu, 233000, China

    Suo-wen Xu

  5. Department of Cardiology, The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, 230036, China

    Ya-jun Duan, Bu-chun Zhang & Hao Hu

  6. College of Life Sciences, Key Laboratory of Bioactive Materials of Ministry of Education, Nankai University, Tianjin, 300071, China

    Ji-hong Han

  7. Diabetes and Obesity Research Center, New York University Long Island School of Medicine, New York, NY, USA

    Qing R. Miao

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  1. Xiao-kang Zhao
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Contributions

Conceptualization, YLC and HH; methodology and investigation, XKZ, MMZ, SNW, TTZ, XNW, CYW, JZ, WYZ, XY, BCZ, and MXJ; resources: SWX; writing—original draft preparation, XKZ and YLC; writing—review and editing, YLC, SWX, YJD, JHH, QRM, and HH; supervision, YLC; funding acquisition, YLC, JHH, MXJ, and BCZ. All authors have read and agreed to the final version of the paper.

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Correspondence to Hao Hu or Yuan-li Chen.

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Zhao, Xk., Zhu, Mm., Wang, Sn. et al. Transcription factor 21 accelerates vascular calcification in mice by activating the IL-6/STAT3 signaling pathway and the interplay between VSMCs and ECs. Acta Pharmacol Sin 44, 1625–1636 (2023). https://doi.org/10.1038/s41401-023-01077-8

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