Identification of miR-34 regulatory networks in settings of disease and antimiR-therapy: Implications for treating cardiac pathology and other diseases

RNA Biol. 2017 May 4;14(5):500-513. doi: 10.1080/15476286.2016.1181251. Epub 2016 Apr 28.


Expression of the miR-34 family (miR-34a, -34b, -34c) is elevated in settings of heart disease, and inhibition with antimiR-34a/antimiR-34 has emerged as a promising therapeutic strategy. Under chronic cardiac disease settings, targeting the entire miR-34 family is more effective than targeting miR-34a alone. The identification of transcription factor (TF)-miRNA regulatory networks has added complexity to understanding the therapeutic potential of miRNA-based therapies. Here, we sought to determine whether antimiR-34 targets secondary miRNAs via TFs which could contribute to antimiR-34-mediated protection. Using miRNA-Seq we identified differentially regulated miRNAs in hearts from mice with cardiac pathology due to transverse aortic constriction (TAC), and focused on miRNAs which were also regulated by antimiR-34. Two clusters of stress-responsive miRNAs were classified as "pathological" and "cardioprotective," respectively. Using ChIPBase we identified 45 TF binding sites on the promoters of "pathological" and "cardioprotective" miRNAs, and 5 represented direct targets of miR-34, with the capacity to regulate other miRNAs. Knockdown studies in a cardiomyoblast cell line demonstrated that expression of 2 "pathological" miRNAs (let-7e, miR-31) was regulated by one of the identified TFs. Furthermore, by qPCR we confirmed that expression of let-7e and miR-31 was lower in hearts from antimiR-34 treated TAC mice; this may explain why targeting the entire miR-34 family is more effective than targeting miR-34a alone. Finally, we showed that Acsl4 (a common target of miR-34, let-7e and miR-31) was increased in hearts from TAC antimiR-34 treated mice. In summary, antimiR-34 regulates the expression of other miRNAs and this has implications for drug development.

Keywords: Heart disease; miR-Seq; miRNA networks; microRNA; therapy.

MeSH terms

  • Adult
  • Analysis of Variance
  • Animals
  • Cardiomegaly / metabolism
  • Cardiomegaly / therapy*
  • Cell Line
  • Coenzyme A Ligases / genetics
  • Coenzyme A Ligases / metabolism
  • Disease Models, Animal
  • Gene Expression Regulation
  • Gene Regulatory Networks*
  • Heart Failure / metabolism
  • Heart Failure / therapy*
  • Heart Ventricles / chemistry
  • Heart Ventricles / metabolism
  • Humans
  • Male
  • Mice
  • Mice, Inbred Strains
  • MicroRNAs / analysis
  • MicroRNAs / antagonists & inhibitors*
  • MicroRNAs / metabolism*
  • Myocytes, Cardiac / chemistry
  • Myocytes, Cardiac / metabolism
  • Placebos
  • Sequence Analysis, RNA
  • Transcription Factors / genetics
  • Transcription Factors / metabolism


  • MIRN34a microRNA, mouse
  • MicroRNAs
  • Placebos
  • Transcription Factors
  • Acsl4 protein, mouse
  • Coenzyme A Ligases