Open AccessmiR‐342‐5p Is a Notch Downstream Molecule and Regulates Multiple Angiogenic Pathways Including Notch, Vascular Endothelial Growth Factor and Transforming Growth Factor β Signaling
Author(s) -
Yan XianChun,
Cao Jing,
Liang Liang,
Wang Li,
Gao Fang,
Yang ZiYan,
Duan JuanLi,
Chang TianFang,
Deng SanMing,
Liu Yuan,
Dou GuoRui,
Zhang Jian,
Zheng QiJun,
Zhang Ping,
Han Hua
Publication year - 2016
Publication title -
journal of the american heart association
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.494
H-Index - 85
ISSN - 2047-9980
DOI - 10.1161/jaha.115.003042
Subject(s) - angiogenesis , notch signaling pathway , vascular endothelial growth factor , smad , vascular endothelial growth factor a , microbiology and biotechnology , transforming growth factor , cancer research , signal transduction , medicine , biology , vegf receptors
Background Endothelial cells ( EC s) form blood vessels through angiogenesis that is regulated by coordination of vascular endothelial growth factor ( VEGF ), Notch, transforming growth factor β, and other signals, but the detailed molecular mechanisms remain unclear. Methods and Results Small RNA sequencing initially identified miR‐342‐5p as a novel downstream molecule of Notch signaling in EC s. Reporter assay, quantitative reverse transcription polymerase chain reaction and Western blot analysis indicated that miR‐342‐5p targeted endoglin and modulated transforming growth factor β signaling by repressing SMAD 1/5 phosphorylation in EC s. Transfection of miR‐342‐5p inhibited EC proliferation and lumen formation and reduced angiogenesis in vitro and in vivo, as assayed by using a fibrin beads–based sprouting assay, mouse aortic ring culture, and intravitreal injection of miR‐342‐5p agomir in P3 pups. Moreover, miR‐342‐5p promoted the migration of EC s, accompanied by reduced endothelial markers and increased mesenchymal markers, indicative of increased endothelial–mesenchymal transition. Transfection of endoglin at least partially reversed endothelial–mesenchymal transition induced by miR‐342‐5p. The expression of miR‐342‐5p was upregulated by transforming growth factor β, and inhibition of miR‐342‐5p attenuated the inhibitory effects of transforming growth factor β on lumen formation and sprouting by EC s. In addition, VEGF repressed miR‐342‐5p expression, and transfection of miR‐342‐5p repressed VEGFR 2 and VEGFR 3 expression and VEGF ‐triggered Akt phosphorylation in EC s. miR‐342‐5p repressed angiogenesis in a laser‐induced choroidal neovascularization model in mice, highlighting its clinical potential. Conclusions miR‐342‐5p acts as a multifunctional angiogenic repressor mediating the effects and interaction among angiogenic pathways.