PMID- 29408314 OWN - NLM STAT- MEDLINE DCOM- 20190916 LR - 20190916 IS - 1600-0641 (Electronic) IS - 0168-8278 (Print) IS - 0168-8278 (Linking) VI - 68 IP - 6 DP - 2018 Jun TI - Fibroblast growth factor receptor inhibition induces loss of matrix MCL1 and necrosis in cholangiocarcinoma. PG - 1228-1238 LID - S0168-8278(18)30073-4 [pii] LID - 10.1016/j.jhep.2018.01.026 [doi] AB - BACKGROUND & AIMS: Myeloid cell leukemia 1 (MCL1), a prosurvival member of the BCL2 protein family, has a pivotal role in human cholangiocarcinoma (CCA) cell survival. We previously reported that fibroblast growth factor receptor (FGFR) signalling mediates MCL1-dependent survival of CCA cells in vitro and in vivo. However, the mode and mechanisms of cell death in this model were not delineated. METHODS: Human CCA cell lines were treated with the pan-FGFR inhibitor LY2874455 and the mode of cell death examined by several complementary assays. Mitochondrial oxidative metabolism was examined using a XF24 extracellular flux analyser. The efficiency of FGFR inhibition in patient-derived xenografts (PDX) was also assessed. RESULTS: CCA cells expressed two species of MCL1, a full-length form localised to the outer mitochondrial membrane, and an N terminus-truncated species compartmentalised within the mitochondrial matrix. The pan-FGFR inhibitor LY2874455 induced non-apoptotic cell death in the CCA cell lines associated with cellular depletion of both MCL1 species. The cell death was accompanied by failure of mitochondrial oxidative metabolism and was most consistent with necrosis. Enforced expression of N terminus-truncated MCL1 targeted to the mitochondrial matrix, but not full-length MCL1 targeted to the outer mitochondrial membrane, rescued cell death and mitochondrial function. LY2874455 treatment of PDX-bearing mice was associated with tumour cell loss of MCL1 and cell necrosis. CONCLUSIONS: FGFR inhibition induces loss of matrix MCL1, resulting in cell necrosis. These observations support a heretofore unidentified, alternative MCL1 survival function, namely prevention of cell necrosis, and have implications for treatment of human CCA. LAY SUMMARY: Herein, we report that therapeutic inhibition of a cell receptor expressed by bile duct cancer cells resulted in the loss of a critical survival protein termed MCL1. Cellular depletion of MCL1 resulted in the death of the cancer cells by a process characterised by cell rupture. Cell death by this process can stimulate the immune system and has implications for combination therapy using receptor inhibition with immunotherapy. CI - Copyright (c) 2018 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved. FAU - Kabashima, Ayano AU - Kabashima A AD - Division of Gastroenterology and Hepatology, Mayo Clinic, Rochester, MN, USA. FAU - Hirsova, Petra AU - Hirsova P AD - Division of Gastroenterology and Hepatology, Mayo Clinic, Rochester, MN, USA; Institute of Clinical Biochemistry and Diagnostics, Charles University, Faculty of Medicine and University Hospital Hradec Kralove, Hradec Kralove, Czech Republic; Department of Pharmacology, Charles University, Faculty of Medicine in Hradec Kralove, Hradec Kralove, Czech Republic. FAU - Bronk, Steven F AU - Bronk SF AD - Division of Gastroenterology and Hepatology, Mayo Clinic, Rochester, MN, USA. FAU - Hernandez, Matthew C AU - Hernandez MC AD - Department of Surgery, Mayo Clinic, Rochester, MN, USA. FAU - Truty, Mark J AU - Truty MJ AD - Department of Surgery, Mayo Clinic, Rochester, MN, USA. FAU - Rizvi, Sumera AU - Rizvi S AD - Division of Gastroenterology and Hepatology, Mayo Clinic, Rochester, MN, USA. FAU - Kaufmann, Scott H AU - Kaufmann SH AD - Department of Molecular Pharmacology and Experimental Therapeutics, Mayo Clinic, Rochester, MN, USA. FAU - Gores, Gregory J AU - Gores GJ AD - Division of Gastroenterology and Hepatology, Mayo Clinic, Rochester, MN, USA. Electronic address: gores.gregory@mayo.edu. LA - eng GR - P30 DK084567/DK/NIDDK NIH HHS/United States GR - R01 CA166741/CA/NCI NIH HHS/United States GR - R01 DK059427/DK/NIDDK NIH HHS/United States GR - R56 DK059427/DK/NIDDK NIH HHS/United States PT - Journal Article PT - Research Support, N.I.H., Extramural PT - Research Support, Non-U.S. Gov't DEP - 20180309 PL - Netherlands TA - J Hepatol JT - Journal of hepatology JID - 8503886 RN - 0 (2-(4-(2-(5-(1-(3,5-dichloropyridin-4-yl)ethoxy)-1H-indazol-3yl)vinyl)-1H-pyrazol-1-yl)ethanol) RN - 0 (A-1210477) RN - 0 (Indazoles) RN - 0 (Indoles) RN - 0 (MCL1 protein, human) RN - 0 (Myeloid Cell Leukemia Sequence 1 Protein) RN - 0 (Receptors, Fibroblast Growth Factor) RN - 0 (Sulfonamides) SB - IM MH - Animals MH - Bile Duct Neoplasms/*drug therapy/*metabolism/pathology MH - Cell Death/drug effects MH - Cell Line, Tumor MH - Cholangiocarcinoma/*drug therapy/*metabolism/pathology MH - Humans MH - Indazoles/*pharmacology MH - Indoles/pharmacology MH - Male MH - Mice MH - Mice, Inbred NOD MH - Mice, SCID MH - Mitochondria/drug effects/metabolism MH - Myeloid Cell Leukemia Sequence 1 Protein/antagonists & inhibitors/*metabolism MH - Necrosis MH - Oxidation-Reduction MH - Receptors, Fibroblast Growth Factor/*antagonists & inhibitors MH - Sulfonamides/pharmacology MH - Xenograft Model Antitumor Assays PMC - PMC5960421 MID - NIHMS939790 OTO - NOTNLM OT - A-1210477 OT - Fibroblast growth factor receptor (FGFR) OT - LY2874455 OT - Patient derived xenograft (PDX) OT - Seahorse extracellular flux analysis COIS- Conflicts of Interest: The authors declare no conflicts of interest with the contents of this article. EDAT- 2018/02/07 06:00 MHDA- 2019/09/17 06:00 PMCR- 2019/06/01 CRDT- 2018/02/07 06:00 PHST- 2017/06/30 00:00 [received] PHST- 2018/01/18 00:00 [revised] PHST- 2018/01/20 00:00 [accepted] PHST- 2018/02/07 06:00 [pubmed] PHST- 2019/09/17 06:00 [medline] PHST- 2018/02/07 06:00 [entrez] PHST- 2019/06/01 00:00 [pmc-release] AID - S0168-8278(18)30073-4 [pii] AID - 10.1016/j.jhep.2018.01.026 [doi] PST - ppublish SO - J Hepatol. 2018 Jun;68(6):1228-1238. doi: 10.1016/j.jhep.2018.01.026. Epub 2018 Mar 9.