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MicroRNA-132–mediated loss of p120RasGAP activates the endothelium to facilitate pathological angiogenesis

Abstract

Although it is well established that tumors initiate an angiogenic switch, the molecular basis of this process remains incompletely understood. Here we show that the miRNA miR-132 acts as an angiogenic switch by targeting p120RasGAP in the endothelium and thereby inducing neovascularization. We identified miR-132 as a highly upregulated miRNA in a human embryonic stem cell model of vasculogenesis and found that miR-132 was highly expressed in the endothelium of human tumors and hemangiomas but was undetectable in normal endothelium. Ectopic expression of miR-132 in endothelial cells in vitro increased their proliferation and tube-forming capacity, whereas intraocular injection of an antagomir targeting miR-132, anti–miR-132, reduced postnatal retinal vascular development in mice. Among the top-ranking predicted targets of miR-132 was p120RasGAP, which we found to be expressed in normal but not tumor endothelium. Endothelial expression of miR-132 suppressed p120RasGAP expression and increased Ras activity, whereas a miRNA-resistant version of p120RasGAP reversed the vascular response induced by miR-132. Notably, administration of anti–miR-132 inhibited angiogenesis in wild-type mice but not in mice with an inducible deletion of Rasa1 (encoding p120RasGAP). Finally, vessel-targeted nanoparticle delivery1 of anti–miR-132 restored p120RasGAP expression in the tumor endothelium, suppressed angiogenesis and decreased tumor burden in an orthotopic xenograft mouse model of human breast carcinoma. We conclude that miR-132 acts as an angiogenic switch by suppressing endothelial p120RasGAP expression, leading to Ras activation and the induction of neovascularization, whereas the application of anti–miR-132 inhibits neovascularization by maintaining vessels in the resting state.

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Figure 1: miR-132 regulates growth factor–mediated angiogenesis in vitro and in vivo.
Figure 2: Endothelial activation mediated by miR-132 depends on its downregulation of p120RasGAP.
Figure 3: miR-132 and p120RasGAP are expressed reciprocally in quiescent versus proliferative endothelium.
Figure 4: Targeted delivery of anti–miR-132 decreases tumor burden by restoring endothelial p120RasGAP.

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Acknowledgements

We thank L. Barnes, E. Goka, B. Walsh and D. Wu for technical support. We thank S. Weng and J. Desgrosellier for discussions. We thank E. Brown (University of Pennsylvania) for the Ert2-ubiquitin-Cre mice. We thank R. Kerbel (University of Toronto) for the fast-growing variant of MDA-MB-231 breast carcinoma cells. We thank S. Kajiji (Scripps Research Institute) for FG human pancreatic adenocarcinoma cells. This work was supported by US National Institutes of Health grants HL078912, CA104898 and CA050286 to D.A.C. and HL096498 to P.D.K. S.A. is supported in part by an American Heart Association postdoctoral fellowship 09POST2040038.

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Contributions

S.A. and D.A.C. designed the study. E.A.M., B.K.M. and R.M. designed the nanoparticles. J.N.L. established the human ES cell vasculogenesis model. D.J.S. helped with the TaqMan microRNA panel experiments and analysis. P.E.L. and P.D.K. generated and characterized the Rasa1fl/fl mice. S.A., L.S., L.M.A. and M.H. performed experiments and analyzed data. S.A., S.M.W. and D.A.C. analyzed data and wrote the manuscript. D.A.C. supervised the project.

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Correspondence to David A Cheresh.

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Anand, S., Majeti, B., Acevedo, L. et al. MicroRNA-132–mediated loss of p120RasGAP activates the endothelium to facilitate pathological angiogenesis. Nat Med 16, 909–914 (2010). https://doi.org/10.1038/nm.2186

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