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Dickkopf 3: a Novel Target Gene of miR-25-3p in Promoting Fibrosis-Related Gene Expression in Myocardial Fibrosis

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Abstract

Increasing evidence has shown that microRNAs (miRNAs) participate in cardiac fibrosis. We aimed to elucidate the effect of miRNA miR-25-3p on cardiac fibrosis. MiRNA microarray was used to profile miRNAs in the myocardium of angiotensin-II (Ang-II)-infused mice. Effect of miR-25-3p on expression of fibrosis-related genes, including Col1a1, Col3a1, and Acta2, was investigated both in vitro and in vivo. MiR-25-3p was shown increased in the myocardium of Ang-II-infused mice and patients with heart failure. MiR-25-3p enhanced fibrosis-related gene expression in mouse cardiac fibroblasts (mCFs) and in the myocardium of Ang-II-infused mice. Dickkopf 3 (Dkk3) was identified as a target gene of miR-25-3p, and Dkk3 could ameliorate Smad3 activation and fibrosis-related gene expression via enhancing Smad7 expression in mCFs. Additionally, NF-κB signal was proven to mediate upregulation of miR-25-3p in cardiac fibrosis. Our findings suggest that miR-25-3p enhances cardiac fibrosis by suppressing Dkk3 to activate Smad3 and fibrosis-related gene expression.

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Abbreviations

miR:

microRNA

Ang-II:

Angiotensin-II

Dkk3:

Dickkopf 3

mCFs:

Mouse cardiac fibroblasts

HF:

Heart failure

TAC:

Transaortic constriction

Smad7:

SMAD family member 7

LV:

Left ventricular

EF:

Ejection fraction

FS:

Fractional shortening

CVF:

Collagen volume fraction

NMVCs:

Neonatal mouse ventricular cardiomyocytes

RT-qPCR:

Quantitative reverse-transcription PCR

FL:

Firefly luciferase

RL:

Renilla luciferase

ChIP:

Chromatin immunoprecipitation

EdU:

5-Ethynyl-2′-deoxyuridine

3′-UTR:

3′-untranslation region

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Acknowledgements

This study was funded by the National Natural Science Foundation of China (82070254 and 81770264 to Z.-X. Shan), a High-level Hospital Construction Project of Guangdong Provincial People’s Hospital (DFJH201902 to Z.-X. Shan), and a Guangzhou Science and Technology Program project (202002030013 to X.-H. Fang).

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Author notes

  1. Ni Zeng and Yi-Hong Wen contributed equally to this work.

Authors and Affiliations

  1. School of Medicine, South China University of Technology, Guangzhou, 510632, China

    Ni Zeng, Yi-Hong Wen & Zhi-Xin Shan

  2. School of Bioscience and Bioengineering, South China University of Technology, Guangzhou, 510632, China

    Rong Pan

  3. Department of Cardiology, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou, 510120, China

    Jing Yang

  4. School of Pharmacy, Southern Medical University, Guangzhou, 510515, China

    Yu-Min Yan, An-Zhi Zhao & Zhi-Xin Shan

  5. Guangdong Provincial Key Laboratory of Clinical Pharmacology, Guangdong Cardiovascular Institute, Guangzhou, 510080, China

    Jie-Ning Zhu & Xian-Hong Fang

  6. Research Center of Medical Sciences, Guangdong Provincial People’s Hospital, Guangdong Academy of Medical Sciences, Guangzhou, 510080, China

    Jie-Ning Zhu & Zhi-Xin Shan

Authors
  1. Ni Zeng
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Contributions

Conceived and designed the experiments: Ni Zeng and Zhi-Xin Shan. Performed the experiments: Ni Zeng, Yi-Hong Wen, Rong Pan, An-Zhi Zhao, Jing Yang, and Yu-Min Yan. Managed data: Ni Zeng and Xian-Hong Fang. Analyzed the data: Yi-Hong Wen, Jie-Ning Zhu, and Zhi-Xin Shan. Contributed reagents/materials/analysis tools: Yu-Min Yan and Xian-Hong Fang. Funding acquisition: Xian-Hong Fang and Zhi-Xin Shan. Wrote the paper: Zhi-Xin Shan. All authors have read and agreed to the final version of manuscript.

Corresponding authors

Correspondence to Xian-Hong Fang or Zhi-Xin Shan.

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Associate Editor Yihua Bei oversaw the review of this article

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Zeng, N., Wen, YH., Pan, R. et al. Dickkopf 3: a Novel Target Gene of miR-25-3p in Promoting Fibrosis-Related Gene Expression in Myocardial Fibrosis. J. of Cardiovasc. Trans. Res. 14, 1051–1062 (2021). https://doi.org/10.1007/s12265-021-10116-w

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