AUTHOR=Myint Kyaw Zwar , Sueca-Comes Mireia , Collier Pamela , Balasubramanian Brinda , Venkatraman Simran , Gordan John , Zaitoun Abed M. , Mukherjee Abhik , Arora Arvind , Larbcharoensub Noppadol , Suriyonplengsaeng Chinnawut , Wongprasert Kanokpan , Janvilisri Tavan , Gomez Dhanny , Grabowska Anna M. , Tohtong Rutaiwan , Bates David O. , Yacqub-Usman Kiren 

TITLE=Preclinical evidence for anaplastic lymphoma kinase inhibitors as novel therapeutic treatments for cholangiocarcinoma

JOURNAL=Frontiers in Oncology

VOLUME=Volume 13 - 2023

YEAR=2023

URL=https://www.frontiersin.org/journals/oncology/articles/10.3389/fonc.2023.1184900

DOI=10.3389/fonc.2023.1184900

ISSN=2234-943X

ABSTRACT=Bile duct cancer (cholangiocarcinoma, CCA), has a poor prognosis for patients, and despite recent advances in targeted therapies for other cancer types, is still treated with standard chemotherapy. Anaplastic Lymphoma Kinase (ALK) has been shown to be a primary driver of disease progression in lung cancer, and ALK inhibitors are effective therapeutics in aberrant ALK expressing tumours. Aberrant ALK expression has been documented in CCA, but the use of ALK inhibitors has not been investigated. Using CCA cell lines and close-to-patient primary cholangiocarcinoma cells we investigated the potential for ALK inhibitors in CCA. Methods: ALK, cMET and ROS1 expression was determined in CCA patient tissue by immunohistochemistry and digital-droplet polymerase chain reaction, in cell lines by immunoblot and immunofluorescence. Effect on cell viability and mechanism of action of ALK, cMet and ROS1 inhibitors was determined in CCA cell lines. To determine whether ceritinib could affect primary CCA cells, tissue was taken from 4 patients with biliary tract cancer, without ALK rearrangement, mutation or overexpression, and grown in three-dimensional tumour growth assays in the presence or absence of humanized mesenchymal cells.ALK and cMet but not ROS were both upregulated in CCA tissues and cell lines. Cell survival was inhibited by crizotinib, a c-met/ALK/ROS inhibitor. To determine the mechanism of this effect, we tested c-Met specific and ALK/ROS specific inhibitors, capmatinib and ceritinib respectively. Whereas capmatinib did not affect cell survival, ceritinib dose-dependently inhibited survival in all cell lines, with IC50 ranging from 1-9 µM and co-treatments with Gemcitabine and Cisplatin further sensitized cells, with IC50 ranging from IC50 0.60-2.32 µM. Ceritinib did not inhibit cMet phosphorylation but did inhibit ALK phosphorylation. ALK was not mutated in any of these cell lines. Only ceritinib inhibited 3D growth of all 4 patient samples below mean peak serum concentration, in the presence and absence of mesenchymal cells, whereas crizotinib and capmatinib failed to do this. Ceritinib appeared to exert its effect more through through autophagy than apoptosis.These results indicate that ceritinib, or other ALK/ROS inhibitors could be therapeutically useful in cholangiocarcinoma even in the absence of aberrant ALK/ROS1 expression.