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Suppression of the Inhibitory Effect of circ_0036176-Translated Myo9a-208 on Cardiac Fibroblast Proliferation by miR-218-5p

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Abstract

Increasing evidence has shown that circular RNAs (circRNAs) participate in the process of cardiac remodeling. CircRNA circ_0036176 originating from the back-splicing of exon 2 to exon4 of myosin IXA (Myo9a) gene was shown to be increased in the myocardium of patients with heart failure (HF) and riched in exosomes from human AC16 cardiomyocytes with overexpression of circ_0036176. Proliferation activity was inhibited in mCFs subjected to exosomal circ_0036176 treatment and in mCFs with overexpression of circ_0036176. Interestingly, circ_0036176 contains an IRES element and an ORF of 627 nt encoding a 208-amino acid protein (termed as Myo9a-208). Myo9a-208 was shown to mediate the inhibitory effect of circ_0036176 on CFs proliferation, and miR-218-5p could inhibit Myo9a-208 expression by binding to circ_0036176, resulting in abolishing the effect of circ_0036176 on inactivating cyclin/Rb signal and suppressing CFs proliferation. Our findings suggest that circ_0036176 inhibits mCFs proliferation by translating Myo9a-208 protein to suppress cyclin/Rb pathway.

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Abbreviations

Ang-II:

Angiotensin-II

CCK8:

Cell Counting Kit-8

CCND1:

Cyclin D1

CCNE1:

Cyclin E1

CDK6:

Cyclin kinase 6

CDK9:

Cyclin kinase 9

circRNAs:

Circular RNAs

CVF:

Collagen volume fraction

ECM:

Extracellular matrix

EdU:

5-Ethynyl-2′-deoxyuridine

FL:

Firefly luciferase

HAFs:

Human atrial fibroblasts

HF:

Heart failure

IRES:

Internal ribosome entry sites

mCFs:

Mouse cardiac fibroblasts

MIRESs:

M6A-induced ribosome engagement sites

miR:

MicroRNA

MYO9A:

Myosin IXA

ORF:

Open reading frame

p-HH3:

Phosphorylated histone H3

p-Rb1:

Phosphorylated RB transcriptional corepressor 1

RL:

Renilla luciferase

RT-qPCR:

Quantitative reverse-transcription PCR

TEM:

Transmission electron microscope

3′-UTR:

3′ Untranslated region

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Funding

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), Guangdong Basic and Applied Basic Research Foundation (2021A1515011554 to X.-H. Fang), and Guangzhou Science and Technology Program project (202002030013 to X.-H. Fang, 202002030039 to H.-M. Guo).

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Author notes

  1. **g Guo, Li-Wen Chen and Zhi-Qi Huang contributed equally to this work.

Authors and Affiliations

  1. Guangdong Provincial Key Laboratory of Clinical Pharmacology, Guangdong Provincial People’s Hospital, Guangdong Academy of Medical Sciences, Guangzhou, 510080, China

    **g Guo, Hui Li, Yu-Min Yan, Jie-Ning Zhu, **an-Hong Fang & Zhi-**n Shan

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

    **g Guo

  3. Guangdong Cardiovascular Institute, Guangzhou, 510080, China

    Li-Wen Chen, Hui-Ming Guo & Zhi-**n Shan

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

    Zhi-Qi Huang

  5. The Second School of Clinical Medicine, Southern Medical University, Guangzhou, 510280, China

    Ji-Shen Guo & Ze-Run Chen

  6. Guangzhou Foreign Language School, Guangzhou, 511455, China

    Yue Shan

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Contributions

Conceived and designed the experiments: **g Guo and Zhi-**n Shan. Performed the experiments: **g Guo, Li-Wen Chen, Zhi-Qi Huang, Ji-Shen Guo, Yue Shan, and Jie-Ning Zhu. Managed data: Zhi-Qi Huang and Hui Li. Analyzed the data: Hui-Ming Guo and Zhi-**n Shan. Contributed reagents/materials/analysis tools: Yu-Min Yan, Ze-Run Chen, and **an-Hong Fang. Funding acquisition: **an-Hong Fang, Hui-Ming Guo, and Zhi-**n Shan. Wrote the paper: Zhi-**n Shan. All authors have read and agreed to the final version of manuscript.

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Correspondence to **an-Hong Fang or Zhi-**n Shan.

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Associate Editor Junjie **ao oversaw the review of this article.

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Guo, J., Chen, LW., Huang, ZQ. et al. Suppression of the Inhibitory Effect of circ_0036176-Translated Myo9a-208 on Cardiac Fibroblast Proliferation by miR-218-5p. J. of Cardiovasc. Trans. Res. 15, 548–559 (2022). https://doi.org/10.1007/s12265-022-10228-x

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