Abstract 4938: Molecular mechanisms of intrinsic resistance to taxanes in gastric cancer

Gastric cancer (GC) is histologically divided into intestinal (INT) and diffuse (DIF) clinical subtypes. Even though these two GC groups are structurally and biologically different, this classification is not used to inform choice of treatment. Taxanes (paclitaxel, docetaxel (DTX) and cabazitaxel) are widely used for cancer treatment and for GC specifically the TAX-325 study revealed therapeutic benefit when DTX was added to the standard chemotherapy regimen. Despite these improvements, however, patients often exhibit intrinsic or acquired resistance to DTX adversely affecting patient survival. Yet, the molecular basis of clinical drug resistance remains poorly elucidated posing a major barrier for the effective treatment of GC patients. We performed a post-hoc analysis of the TAX-325 study to examine the potential influence of GC subtypes in clinical response to DTX. We classified randomized patients as diffuse or non-diffuse histology and correlated histology with clinical outcomes using a Cox proportional hazards model. Non-diffuse GC showed a significant improvement in overall survival with the addition of DTX (12.1 v 8.8 mo, p = 0.002), whereas diffuse histology was not associated with an improvement in survival (8.3 v 8.5 mo, p = 0.66). To investigate the molecular mechanism of GC DIF subtype resistance to taxanes, we used a panel of 12 GC cell lines representative of both subtypes (4 intestinal subtype, 8 diffuse subtype). DTX cytotoxicity assays revealed that similarly to what we observed clinically, 63% (5/8) of the DIF GC cell lines were resistant (IC50 > 600 nM) to DTX compared to 25% (1/4) of the INT GC cell lines. Further functional studies revealed that there was minimal DTX drug-target engagement in the DIF GC cells, assessed by confocal microscopy of the microtubule (MT) network and tubulin polymerization assays. These results suggested that DTX interaction with its target, MT, was impaired in the DIF GC cell lines. To rule out multi-drug resistance (MDR) as potential cause of intrinsic DTX resistance in these cells we performed flow cytometric evaluation of P-glycoprotein and found that all of the DIF GC cell lines were negative. Additionally, drug accumulation studies with C-14 radiolabeled DTX revealed that the drug accumulated intracellularly in all of GC cell lines in our panel. Next generation sequencing of our panel of untreated or DTX-treated GC cell lines revealed 84 significantly differentially expressed genes in drug-sensitive cell lines but no changes in resistant cells. Our analysis showed a significant enrichment and a transcriptional co-regulation after treatment of genes encoding for kinesins, motor proteins associated with MT, in DTX-sensitive cells but not in DTX-resistant cells. These studies will provide novel insights into mechanism of drug resistance and sensitivity and will ultimately allow us to design more effective targeted therapies to overcome chemo-resistance and eventually prolong patient survival. Citation Format: Giuseppe Galletti, Chao Zhang, Kyle Cleveland, Doron Betel, Manish Shah, Paraskevi Giannakakou. Molecular mechanisms of intrinsic resistance to taxanes in gastric cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4938.