title: Autophagy is involved in endogenous and NVP-AUY922-induced KIT degradation in gastrointestinal stromal tumors.
abstract: Gastrointestinal stromal tumor (GIST) is a prototype of mutant KIT oncogene-driven tumor. Prolonged tyrosine kinase inhibitor (TKI) treatment may result in a resistant phenotype through acquired secondary KIT mutation. Heat shock protein 90 (HSP90AA1) is a chaperone protein responsible for protein maturation and stability, and KIT is a known client protein of HSP90AA1. Inhibition of HSP90AA1 has been shown to destabilize KIT protein by enhancing its degradation via the proteasome-dependent pathway. In this study, we demonstrated that NVP-AUY922 (AUY922), a new class of HSP90AA1 inhibitor, is effective in inhibiting the growth of GIST cells expressing mutant KIT protein, the imatinib-sensitive GIST882 and imatinib-resistant GIST48 cells. The growth inhibition was accompanied with a sustained reduction of both total and phosphorylated KIT proteins and the induction of apoptosis in both cell lines. Surprisingly, AUY922-induced KIT reduction could be partially reversed by pharmacological inhibition of either autophagy or proteasome degradation pathway. The blockade of autophagy alone led to the accumulation of the KIT protein, highlighting the role of autophagy in endogenous KIT turnover. The involvement of autophagy in endogenous and AUY922-induced KIT protein turnover was further confirmed by the colocalization of KIT with MAP1LC3B-, acridine orange- or SQSTM1-labeled autophagosome, and by the accumulation of KIT in GIST cells by silencing either BECN1 or ATG5 to disrupt autophagosome activity. Therefore, the results not only highlight the potential application of AUY922 for the treatment of KIT-expressing GISTs, but also provide the first evidence for the involvement of autophagy in endogenous and HSP90AA1 inhibitor-induced KIT degradation.
authors.affiliation: Institute of Clinical Pharmacy and Pharmaceutical Science, National Cheng Kung University, Tainan, Taiwan.
authors.forename: Yuan-Shuo
authors.initials: YS
authors.lastname: Hsueh
authors.forename: Chueh-Chuan
authors.initials: CC
authors.lastname: Yen
authors.forename: Neng-Yao
authors.initials: NY
authors.lastname: Shih
authors.forename: Nai-Jung
authors.initials: NJ
authors.lastname: Chiang
authors.forename: Chien-Feng
authors.initials: CF
authors.lastname: Li
authors.forename: Li-Tzong
authors.initials: LT
authors.lastname: Chen
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology; D001343:Autophagy / Q000187:drug effects*; D001549:Benzamides / Q000494:pharmacology; D016227:Benzoquinones / Q000494:pharmacology; D016923:Cell Death / Q000187:drug effects; D002457:Cell Extracts; D045744:Cell Line, Tumor; D015536:Down-Regulation / Q000187:drug effects; D046152:Gastrointestinal Stromal Tumors / Q000473:pathology*; D006801:Humans; D000068877:Imatinib Mesylate; D007555:Isoxazoles / Q000494:pharmacology*; D047029:Lactams, Macrocyclic / Q000494:pharmacology; D008954:Models, Biological; D010766:Phosphorylation / Q000187:drug effects; D010879:Piperazines / Q000494:pharmacology; D059748:Proteolysis / Q000187:drug effects*; D019009:Proto-Oncogene Proteins c-kit / Q000378:metabolism*; D011743:Pyrimidines / Q000494:pharmacology; D012118:Resorcinols / Q000494:pharmacology*
chemical_list: C528044:5-(2,4-dihydroxy-5-isopropylphenyl)-4-(4-morpholin-4-ylmethylphenyl)isoxazole-3-carboxylic acid ethylamide; D000970:Antineoplastic Agents; D001549:Benzamides; D016227:Benzoquinones; D002457:Cell Extracts; D007555:Isoxazoles; D047029:Lactams, Macrocyclic; D010879:Piperazines; D011743:Pyrimidines; D012118:Resorcinols; C112765:tanespimycin; D000068877:Imatinib Mesylate; D019009:Proto-Oncogene Proteins c-kit