A carvedilol-responsive microRNA, miR-125b-5p protects the heart from acute myocardial infarction by repressing pro-apoptotic bak1 and klf13 in cardiomyocytes

https://doi.org/10.1016/j.yjmcc.2017.11.003Get rights and content

Highlights

  • MiR-125b-5p protects the heart against myocardial infarction.

  • MiR-125b-5p functions as a gatekeeper of cardiomyocyte survival.

  • The action of miR-125b-5p is mediated by the repression of bak1 and klf13.

Abstract

Background

Cardiac injury is accompanied by dynamic changes in the expression of microRNAs (miRs), small non-coding RNAs that post-transcriptionally regulate target genes. MiR-125b-5p is downregulated in patients with end-stage dilated and ischemic cardiomyopathy, and has been proposed as a biomarker of heart failure. We previously reported that the β-blocker carvedilol promotes cardioprotection via β-arrestin-biased agonism of β1-adrenergic receptor while stimulating miR-125b-5p processing in the mouse heart. We hypothesize that β1-adrenergic receptor/β-arrestin1-responsive miR-125b-5p confers the improvement of cardiac function and structure after acute myocardial infarction.

Methods and results

Using cultured cardiomyocyte (CM) and in vivo approaches, we show that miR-125b-5p is an ischemic stress-responsive protector against CM apoptosis. CMs lacking miR-125b-5p exhibit increased susceptibility to stress-induced apoptosis, while CMs overexpressing miR-125b-5p have increased phospho-AKT pro-survival signaling. Moreover, we demonstrate that loss-of-function of miR-125b-5p in the mouse heart causes abnormalities in cardiac structure and function after acute myocardial infarction. Mechanistically, the improvement of cardiac function and structure elicited by miR-125b-5p is in part attributed to repression of the pro-apoptotic genes Bak1 and Klf13 in CMs.

Conclusions

In conclusion, these findings reveal a pivotal role for miR-125b-5p in regulating CM survival during acute myocardial infarction.

Section snippets

Subject codes

Non-coding RNAs

Cell signaling/signal transduction

Receptor pharmacology

Heart failure-basic studies

Animal study approval

Eight to 12-week-old C57BL/6 wild-type (WT) mice and 1- to 2-day-old Sprague-Dawley rats were used for this study. Research with animals carried out for this study was performed according to approved protocols and animal welfare regulations of Augusta University's Institutional IACUC Committees. All animal procedures were performed in accordance with NIH guidelines. Neonatal rats were euthanized by decapitation under anesthesia for CM isolation, and mice were euthanized by thoracotomy with 1–4%

In vivo knockdown of miR-125b-5p results in enhanced post-AMI mortality and left ventricular dysfunction

To investigate the role of miR-125b-5p in experimental MI, we intramyocardially injected LNA™-antimiR-125b-5p into WT mice immediately after LAD occlusion or sham surgery. First, we demonstrated efficacy of the antimiR-125b-5p by showing that the level of miR-125b-5p was reduced, for instance, by ~ 75% after 7 days compared with anti-miR controls in both the sham and MI groups (Fig. 1A and data not shown). We further showed that the hearts of antimiR-125b-5p-injected mice at baseline were

Discussion

Here, we identify miR-125b-5p as an ischemic stress-responsive protector against CM apoptosis both in vivo and in vitro. Knockdown of miR-125b-5p renders mice more sensitive to ischemic injury, as evidenced by increased cardiac apoptosis and fibrosis as well as impairment of ventricular function following AMI. Mechanistically, we determined that miR-125b-5p targets pro-apoptotic bak1 and klf13 to elicit its protective effects. CMs deficient in miR-125b-5p exhibit increased sensitivity to

Conclusions

Our results suggest that miR-125b-5p protects the heart against AMI by blunting CM death in response to injury in part through its repression of bak1 and klf13 (Fig. 8D). Although additional mechanistic studies concentrating on miR-125b-5p in different injury models and in other cardiac cell types are needed, our data nevertheless suggest that boosting miR-125b-5p levels to attenuate CM death may provide therapeutic benefits given that downregulation of miR-125b-5p is associated with ischemic

Sources of funding

This work was supported by American Heart Association Predoctoral Fellowship 16PRE30210016 to Jian-peng Teoh, National Institutes of Health R01 HL086555 to Yaoliang Tang, National Institutes of Health R01 HL124248 to Huabo Su, National Institutes of Health R01 HL134354 and AR070029 to Yaoliang Tang and Neal L. Weintraub, National Institutes of Health R01 HL112640 and HL126949 to Neal L. Weintraub, and American Physiological Society (APHYS00008) Shih-Chun Wang Young Investigator Award, American

Disclosures

The authors declare no conflict of interest.

Acknowledgements

We thank Drs. Ruth Caldwell, Zsolt Bagi and Zheng Dong for sharing their equipment, and Dr. Zuzana Bologna for excellent technical assistance.

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  • Cited by (0)

    1

    Present address: Washington University, Saint Louis, MO, USA.

    2

    Present address: University of Kentucky, Lexington, KY, USA.

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