PUBLICATION

Dysregulated miR-125a promotes angiogenesis through enhanced glycolysis

Authors
Wade, S.M., Ohnesorge, N., McLoughlin, H., Biniecka, M., Carter, S.P., Trenkman, M., Cunningham, C.C., McGarry, T., Canavan, M., Kennedy, B.N., Veale, D.J., Fearon, U.
ID
ZDB-PUB-190902-3
Date
2019
Source
EBioMedicine   47: 402-413 (Journal)
Registered Authors
Kennedy, Breandan N.
Keywords
Angiogenesis, CRISPR/CAS9, Metabolism, Zebrafish, microRNA
MeSH Terms
  • Animals
  • Biopsy
  • Cell Movement
  • Cell Proliferation
  • Disease Models, Animal
  • Endothelial Cells
  • Gene Expression Regulation*
  • Gene Silencing
  • Glycolysis
  • Humans
  • MicroRNAs/genetics*
  • Neovascularization, Pathologic/genetics*
  • Osteoarthritis/genetics
  • Osteoarthritis/pathology
  • RNA Interference
  • Zebrafish
PubMed
31466915 Full text @ EBioMedicine
Abstract
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.
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.
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.
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).
Genes / Markers
Figures
Show all Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping