EBioMedicine (Sep 2019)

Dysregulated miR-125a promotes angiogenesis through enhanced glycolysisResearch in context

  • Sarah M. Wade,
  • Nils Ohnesorge,
  • Hayley McLoughlin,
  • Monika Biniecka,
  • Steven P. Carter,
  • Michelle Trenkman,
  • Clare C. Cunningham,
  • Trudy McGarry,
  • Mary Canavan,
  • Breandán N. Kennedy,
  • Douglas J. Veale,
  • Ursula Fearon

Journal volume & issue
Vol. 47
pp. 402 – 413

Abstract

Read online

Background: Although neoangiogenesis is a hallmark of chronic inflammatory diseases such as inflammatory arthritis and many cancers, therapeutic agents targeting the vasculature remain elusive. Here we identified miR-125a as an important regulator of angiogenesis. Methods: MiRNA levels were quantified in Psoriatic Arthritis (PsA) synovial-tissue by RT-PCR and compared to macroscopic synovial vascularity. HMVEC were transfected with anti-miR-125a and angiogenic mechanisms quantified using tube formation assays, transwell invasion chambers, wound repair, RT-PCR and western blot. Real-time analysis of EC metabolism was assessed using the XF-24 Extracellular-Flux Analyzer. Synovial expression of metabolic markers was assessed by immunohistochemistry and immunofluorescent staining. MiR-125a CRISPR/Cas9-based knock-out zebrafish were generated and vascular development assessed. Finally, glycolytic blockade using 3PO, which inhibits Phosphofructokinase-fructose-2,6-bisphophatase 3 (PFKFB3), was assessed in miR-125a−/− ECs and zebrafish embryos. Findings: MiR-125a is significantly decreased in PsA synovium and inversely associated with macroscopic vascularity. In-vivo, CRISPR/cas9 miR-125a−/− zebrafish displayed a hyper-branching phenotype. In-vitro, miR-125a inhibition promoted EC tube formation, branching, migration and invasion, effects paralleled by a shift in their metabolic profile towards glycolysis. This metabolic shift was also observed in the PsA synovial vasculature where increased expression of glucose transporter 1 (GLUT1), PFKFB3 and Pyruvate kinase muscle isozyme M2 (PKM2) were demonstrated. Finally, blockade of PFKFB3 significantly inhibited anti-miR-125a-induced angiogenic mechanisms in-vitro, paralleled by normalisation of vascular development of CRISPR/cas9 miR-125a−/− zebrafish embryos. Intepretation: Our results provide evidence that miR-125a deficiency enhances angiogenic processes through metabolic reprogramming of endothelial cells. Fund: Irish Research Council, Arthritis Ireland, EU Seventh Framework Programme (612218/3D-NET). Keywords: Angiogenesis, Metabolism, microRNA, CRISPR/CAS9, Zebrafish