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Endothelial cell morphology needs to be properly regulated in angiogenesis. Vascular endothelial growth factor (VEGF) induces endothelial cell elongation, which promotes sprouting of preexisting vessels. However, therapeutic angiogenesis using VEGF has been hampered by side effects such as elevated vascular permeability. Here, we attempted to induce endothelial cell elongation without an overdose of VEGF. By screening a library of chemical inhibitors, we identified phosphatidylinositol 3-kinase (PI3K)/Akt inhibitors and mammalian target of rapamycin complex 1 (mTORC1) inhibitors as potent inducers of endothelial cell elongation. The elongation required VEGF at a low concentration, which was insufficient to elicit the same effect by itself. The elongation also depended on Foxo1, a transcription factor indispensable for angiogenesis. Interestingly, the Foxo1 dependency of the elongation was overridden by inhibition of mTORC1, but not PI3K/Akt, under stimulation by a high concentration of VEGF. Dual inhibition of mTORC1 and mTORC2 failed to induce cell elongation, revealing mTORC2 as a positive regulator of elongation. Our findings suggest that the PI3K/Akt/Foxo1 and mTORC1/mTORC2 pathways differentially regulate endothelial cell elongation, depending on the microenvironmental levels of VEGF.

Figure: Regulation of endothelial cell elongation.
Endothelial cell elongation usually requires a strong stimulation with VEGF. However, inhibition of either the PI3K/Akt or mTORC1 signaling pathway induces the elongation even under a weak stimulation with VEGF.