ALK is a tyrosine kinase receptor involved in a broad range of solid and haematological tumors. It has been shown that about 70-80% of ALK+ Anaplastic Large Cell Lymphoma (ALCL) cases, an aggressive Non Hodgkin T-cell lymphoma, are characterized by the traslocation t(2;5), originating the functional, aberrant oncogenic fusion protein NPM-ALK. Crizotinib was the first clinically relevant ALK inhibitor, now approved for the treatment of late stage and metastatic cases of lung cancer. Several patients soon developed Crizotinib resistance, mainly due to the appearance of new point mutations located in ALK kinase domain. Currently, other ALK inhibitors are available and already in clinical trial, hopefully representing the second line therapy able to overcome Crizotinib resistance. In this work we focused our attention on the more potent phase I/II dual ALK/EGFR inhibitor AP26113 (Ariad Pharm.), able to overcome the Crizotinib-resistant gatekeeper mutation L1196M. Two NPM-ALK+ human cell lines, KARPAS-299 and SUP-M2, were grown in four independent flasks in the presence of increasing doses of AP26113, so that eight cell lines able to grow in the presence of high AP26113 doses were selected. All cell lines show an AP26113 50% inhibitory concentration (IC50) value substantially higher than the one observed in parental cells, with a 130 to 1000-fold increase. All KARPAS-299 populations resistant to AP26113 show NPM-ALK overexpression, due to oncogene amplification, as the main cause of resistance, while SUP-M2 cells proliferating in the presence of the drug harbour several point mutations spanning the entire ALK kinase domain. In particular, we identified the following aminoacid substitutions: L1122V, , L1196M, S1206C and a double F1174V+L1198F mutation. The knowledge of the possible appearance of new clinically relevant mechanisms of drug resistance is a useful tool for the management of new TKI resistant cases
Ceccon, M., Mologni, L., Giudici, G., Piazza, R., Pirola, A., Fontana, D., et al. (2014). Mechanisms of resistance to the second-generation ALK inhibitor AP26113 in human NPM-ALK-positive anaplastic large cell lymphoma cells. In Proceedings: AACR Annual Meeting 2014; April 5-9, 2014; San Diego, CA [10.1158/1538-7445.AM2014-3719].
Mechanisms of resistance to the second-generation ALK inhibitor AP26113 in human NPM-ALK-positive anaplastic large cell lymphoma cells
Ceccon, M;Mologni, L;Piazza, R;Pirola, A;Fontana, D;Gambacorti-Passerini, C
2014
Abstract
ALK is a tyrosine kinase receptor involved in a broad range of solid and haematological tumors. It has been shown that about 70-80% of ALK+ Anaplastic Large Cell Lymphoma (ALCL) cases, an aggressive Non Hodgkin T-cell lymphoma, are characterized by the traslocation t(2;5), originating the functional, aberrant oncogenic fusion protein NPM-ALK. Crizotinib was the first clinically relevant ALK inhibitor, now approved for the treatment of late stage and metastatic cases of lung cancer. Several patients soon developed Crizotinib resistance, mainly due to the appearance of new point mutations located in ALK kinase domain. Currently, other ALK inhibitors are available and already in clinical trial, hopefully representing the second line therapy able to overcome Crizotinib resistance. In this work we focused our attention on the more potent phase I/II dual ALK/EGFR inhibitor AP26113 (Ariad Pharm.), able to overcome the Crizotinib-resistant gatekeeper mutation L1196M. Two NPM-ALK+ human cell lines, KARPAS-299 and SUP-M2, were grown in four independent flasks in the presence of increasing doses of AP26113, so that eight cell lines able to grow in the presence of high AP26113 doses were selected. All cell lines show an AP26113 50% inhibitory concentration (IC50) value substantially higher than the one observed in parental cells, with a 130 to 1000-fold increase. All KARPAS-299 populations resistant to AP26113 show NPM-ALK overexpression, due to oncogene amplification, as the main cause of resistance, while SUP-M2 cells proliferating in the presence of the drug harbour several point mutations spanning the entire ALK kinase domain. In particular, we identified the following aminoacid substitutions: L1122V, , L1196M, S1206C and a double F1174V+L1198F mutation. The knowledge of the possible appearance of new clinically relevant mechanisms of drug resistance is a useful tool for the management of new TKI resistant cases
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