MicroRNA-10a/10b represses a novel target gene mib1 to regulate angiogenesis

Cardiovasc Res. 2016 May 1;110(1):140-50. doi: 10.1093/cvr/cvw023. Epub 2016 Jan 29.

Abstract

Aims: MicroRNA-10 (miR-10) was originally shown to regulate angiogenesis by directly modulating the levels of membrane-bound fms-related tyrosine kinase 1 (mflt1) and its soluble splice isoform sflt1 post-transcriptionally in zebrafish. Given that flt1 knockdown incompletely rescues the angiogenic phenotypes in miR-10 morphants, flt1 is unlikely to be the only important target of miR-10 in endothelial cells (ECs). It will be interesting to investigate new mechanism responsible for angiogenic defect induced by miR-10 knockdown.

Methods and results: Firstly, we demonstrated that miR-10a and miR-10b (miR-10a/10b) were highly enriched in embryonic zebrafish ECs using deep sequencing, Taqman polymerase chain reaction, and in situ hybridisation. Subsequently, we proved that loss of miR-10a/10b impaired blood vessel outgrowth through regulating tip cell behaviours. Mib1 was identified as a potential direct target of miR-10a/10b through in silicon analysis and in vitro luciferase sensor assay. In vivo reporter assay in zebrafish embryos confirmed the binding of miR-10 with 3'-UTR of zebrafish mib1. Furthermore, inhibition of mib1 and Notch signaling rescued the angiogenic defects in miR-10-deficient zebrafish embryos. In addition, we provided evidences that miR-10 regulates human ECs behaviour through targeting Mib1 as well.

Conclusion: Taken together, these results indicate that miR-10 regulates the angiogenic behaviour in a Notch-dependent manner by directly targeting mib1.

Keywords: Notch; angiogenesis; miR-10a; miR-10b; mib1.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Proliferation / genetics*
  • Cell Proliferation / physiology
  • Endothelial Cells / metabolism*
  • Humans
  • MicroRNAs / genetics*
  • MicroRNAs / metabolism
  • Morphogenesis / physiology
  • Neovascularization, Physiologic*
  • Oligonucleotide Array Sequence Analysis / methods
  • Phenotype
  • Ubiquitin-Protein Ligases / genetics*
  • Ubiquitin-Protein Ligases / metabolism*
  • Zebrafish
  • Zebrafish Proteins / genetics*

Substances

  • MIRN10 microRNA, human
  • MIRN10 microRNA, zebrafish
  • MicroRNAs
  • Zebrafish Proteins
  • MIB1 ligase, human
  • Ubiquitin-Protein Ligases
  • mib1 protein, zebrafish