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
Citation
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. (2019) Dysregulated miR-125a promotes angiogenesis through enhanced glycolysis. EBioMedicine. 47:402-413.
Abstract
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).
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping