Abstract 3738: Crizotinib induces tumor suppressor miR-146a, autophagy and p21 waf1/cip1 by alternative targeting non-TK target Akt-mTOR-Foxo axis: A CPI-based systemic study on discovery of phenotypic off-target
Authors: Sun Jiazhi , Yang Lun , Li Minghua , Heller Ian E. , Raplee Issac , Guzman Steven , Pasciak John P. , Cheng Feng , Sneed Kevin B. , He Lin , Zhou Shu-Feng 
Published: May 2015

Abstract
Crizotinib, a first ever ALK /MET multi-targeted receptor tyrosine kinase inhibitor developed by Pfizer and fast approved by FDA in 2011 for the treatment of ALK-rearranged NSCLC patients. We hypothesize that crizotinib acts on other molecular targets in addition to tyrosine kinases ALK/MET and may play critical role given that there is a complex network of kinases that work together to regulate a number of important cellular processes and different disease signature. Approaching with a comprehensive docking method with our established chemical-protein interactome (CPI) and crizotinib, we have discovered 301 PDB-deposited proteins corresponding to 353 ligand binding pockets among a total of 1,780 PDB-deposited human protein entries. Systemic pharmacology approach also applied including validation the molecular target(s) of TKIs in vitro. Especially multiple human cellular models (including NSCLC and melanoma) and signaling study, luc-reporter, autophagy, apoptosis assay, FACS and NGS analysis applied. Interestingly, crizotinib besides had a high CPI binding score (ZZ_score) of -2.2778 against tyrosine kinase Lck, also had -1.4672, -1.1242, -0.7033 and -1.5384 against MAPK3, S6K, GSK-3β and HDAC 7A, respectively, which indicated crizotinib had potential broad range interaction with both TK and ser/thr kinases. In addition, those crizotinib achieved high ZZ scores against PPAR-γ (-1.6145) and VitD3R (0.9479), suggesting a high binding affinity of crizotinib with these nuclear proteins. Our preliminary studies have showed that the crizotinib induced autophagy by activated LC3 in vitro and inhibited oncogenic Akt/mTOR signaling in human multiple melanoma cells resulting in G2/M cell cycle arrest. Furthermore, crizotinib increased p21Waf1/cip1 and Foxo3A expression and tumor suppressor miR-146a in human melanoma cells. Taken together, crizotinib induces miR-146a, autophagy and p21 waf1/cip1 may potentially treat other type of human cancer by concurrently targeting tyrosine kinase, Akt/mTOR/Foxo axis and nuclear receptor, shedding a light for future both anti-cancer and anti-metabolic disorder drug discovery and development. This is the first CPI-based systemic study on leading discovery of crizotinib off-target phenotypic importance. These findings suggest that non-tyrosine kinase target of crizotinib may play critical role(s) for drug repositioning may represent a new and effective approach to expanding the application of existing drugs.
Citation Format: Jiazhi Sun, Lun Yang, Minghua Li, Ian E. Heller, Issac Raplee, Steven Guzman, John P. Pasciak, Feng Cheng, Kevin B. Sneed, Lin He, Shu-Feng Zhou. Crizotinib induces tumor suppressor miR-146a, autophagy and p21 waf1/cip1 by alternative targeting non-TK target Akt-mTOR-Foxo axis: A CPI-based systemic study on discovery of phenotypic off-target. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3738. doi:10.1158/1538-7445.AM2014-3738