Crizotinib Inhibits Metabolic Inactivation of Gemcitabine in c-Met–driven Pancreatic Carcinoma

Abstract

Pancreatic ductal adenocarcinoma (PDAC) remains a major unsolved health problem. Most drugs that pass preclinical tests fail in these patients, emphasizing the need of improved preclinical models to test novel anticancer strategies. Here, we developed four orthotopic mouse models using primary human PDAC cells genetically engineered to express firefly-and Gaussia luciferase, simplifying the ability to monitor tumor growth and metastasis longitudinally in individual animals with MRI and high-frequency ultrasound. In these models, we conducted detailed histopathologic and immunohistochemical analyses on paraffin-embedded pancreatic tissues and metastatic lesions in liver, lungs, and lymph nodes. Genetic characteristics were compared with the originator tumor and primary tumor cells using array-based comparative genomic hybridization, using frozen specimens obtained by laser microdissection. Notably, the orthotopic human xenografts in these models recapitulated the phenotype of human PDACs, including hypovascular and hypoxic areas. Pursuing genomic and immunohistochemical evidence revealed an increased copy number and overexpression of c-Met in one of the models; we examined the preclinical efficacy of c-Met inhibitors in vitro and in vivo. In particular, we found that crizotinib decreased tumor dimension, prolonged survival, and increased blood and tissue concentrations of gemcitabine, synergizing with a cytidine deaminase-mediated mechanism of action. Together, these more readily imaged orthotopic PDAC models displayed genetic, histopathologic, and metastatic features similar to their human tumors of origin. Moreover, their use pointed to c-Met as a candidate therapeutic target in PDAC and highlighted crizotinib and gemcitabine as a synergistic combination of drugs warranting clinical evaluation for PDAC treatment. Cancer Res; 73(22); 6745-56. Ó2013 AACR.

Tissue distribution of crizotinib and gemcitabine combination in a patient-derived orthotopic mouse model of pancreatic cancer

“…Usually drug interactions of combinations receive limited interest. Earlier we investigated the interaction between gemcitabine and crizotinib (Figure 1) and demonstrated a synergistic effect between the drugs in in vitro and in vivo models of pancreatic ductal adenocarcinoma (PDAC) [3]. In the present study we aimed to analyze their interaction in normal tissues, in order to determine whether this information can give an explanation for toxic side effects seen with these drugs in patients.…”

“…Orthotopic primary-PDAC mouse models were developed as reported by Avan et al [3] by implantation of primary tumor cells into 6-8 week old female athymic nude mice (Harlan, Host, The Netherlands). Tumor growth was monitored via bioluminescence, as described previously [3].…”

Development of bioluminescent chick chorioallantoic membrane (CAM) models from primary pancreatic cancer cells: A platform for drug testing

“…These methods could potentially be complemented with bioluminescence (BLI), which is a low-cost longitudinal imaging method. We have successfully developed orthotopic mouse models employing primary human cancer cells genetically engineered to express Firefly-luciferase (Fluc), providing an ease-of-use, low cost and high-throughput imaging mechanism to monitor tumor growth 13,14 .…”

“…These methods could potentially be complemented with bioluminescence (BLI), which is a low-cost longitudinal imaging method. We have successfully developed orthotopic mouse models employing primary human cancer cells genetically engineered to express Firefly-luciferase (Fluc), providing an ease-of-use, low cost and high-throughput imaging mechanism to monitor tumor growth 13,14 .The aim of the present study was to develop CAM bioluminescent tumor models employing low passage cell cultures obtained from primary PDACs, transduced with lentivirus expressing Fluc. Finally, for a pilot pharmacological study, we treated the CAM tumors with gemcitabine, a standard chemotherapeutic agent used for the treatment of PDAC patients, and with the targeted agent crizotinib, that has been previously shown to interact synergistically with gemcitabine 13 .…”

Low expression of chloride channel accessory 1 predicts a poor prognosis in colorectal cancer: The question is still open

“…8,9 We thank Yang et al for their worthwhile findings, but believe that additional parameters and analysis with the incorporation of proper methodology, larger populations, and successive powered statistical analysis are essential to validate the candidate biomarker beyond already available clinical factors for the clinical management of patients with CRC.…”