Simultaneous targeting of NPC1 and VDAC1 by itraconazole leads to synergistic inhibition of MTOR signaling and angiogenesis Journal Article
| Authors: | Head, S. A.; Shi, W. Q.; Yang, E. J.; Nacev, B. A.; Hong, S. Y.; Pasunooti, K. K.; Li, R. J.; Shim, J. S.; Liu, J. O. |
| Article Title: | Simultaneous targeting of NPC1 and VDAC1 by itraconazole leads to synergistic inhibition of MTOR signaling and angiogenesis |
| Abstract: | The antifungal drug itraconazole was recently found to exhibit potent antiangiogenic activity and has since been repurposed as an investigational anticancer agent. Itraconazole has been shown to exert its antiangiogenic activity through inhibition of the mTOR signaling pathway, but the molecular mechanism of action was unknown. We recently identified the mitochondrial protein VDAC1 as a target of itraconazole and a mediator of its activation of AMPK, an upstream regulator of mTOR. However, VDAC1 could not account for the previously reported inhibition of cholesterol trafficking by itraconazole, which was also demonstrated to lead to mTOR inhibition. In this study, we demonstrate that cholesterol trafficking inhibition by itraconazole is due to direct inhibition of the lysosomal protein NPC1. We further map the binding site of itraconazole to the sterol-sensing domain of NPC1 using mutagenesis, competition with U18666A, and molecular docking. Finally, we demonstrate that simultaneous AMPK activation and cholesterol trafficking inhibition leads to synergistic inhibition of mTOR, endothelial cell proliferation, and angiogenesis. © 2016 American Chemical Society. |
| Keywords: | signal transduction; carrier protein; angiogenesis inhibitor; cell proliferation; metabolism; antifungal agent; membrane glycoproteins; carrier proteins; membrane protein; cholesterol; molecular docking; angiogenesis inhibitors; antifungal agents; biological transport; itraconazole; transport at the cellular level; human umbilical vein endothelial cells; hydroxymethylglutaryl coenzyme a reductase kinase; tor serine-threonine kinases; target of rapamycin kinase; umbilical vein endothelial cell; mtor protein, human; drug effects; humans; human; antagonists and inhibitors; npc1 protein, human; vdac1 protein, human; voltage dependent anion channel 1; amp-activated protein kinases; molecular docking simulation; voltage-dependent anion channel 1 |
| Journal Title: | ACS Chemical Biology |
| Volume: | 12 |
| Issue: | 1 |
| ISSN: | 1554-8929 |
| Publisher: | American Chemical Society |
| Date Published: | 2017-01-20 |
| Start Page: | 174 |
| End Page: | 182 |
| Language: | English |
| DOI: | 10.1021/acschembio.6b00849 |
| PUBMED: | 28103683 |
| PROVIDER: | scopus |
| PMCID: | PMC5791891 |
| DOI/URL: | |
| Notes: | Article -- Export Date: 3 April 2017 -- Source: Scopus |
