The Neuroblastoma-Associated F1174L ALK Mutation Causes Resistance to an ALK Kinase Inhibitor in ALK-Translocated Cancers

Abstract

The ALK kinase inhibitor crizotinib (PF-02341066) is clinically effective in patients with ALK-translocated cancers, but its efficacy will ultimately be limited by acquired drug resistance. Here we report the identification of a secondary mutation in ALK, F1174L, as one cause of crizotinib resistance in a patient with an inflammatory myofibroblastic tumor (IMT) harboring a RANBP2-ALK translocation who progressed while on crizotinib therapy. When present in cis with an ALK translocation, this mutation (also detected in neuroblastomas) causes an increase in ALK phosphorylation, cell growth, and downstream signaling. Furthermore, the F1174L mutation inhibits crizotinib-mediated downregulation of ALK signaling and blocks apoptosis in RANBP2-ALK Ba/F3 cells. A chemically distinct ALK inhibitor, TAE684, and the HSP90 inhibitor 17-AAG are both effective in models harboring the F1174L ALK mutation. Our findings highlight the importance of studying drug resistance mechanisms in order to develop effective clinical treatments for patients with ALK-translocated cancers.

Treatment Efficacy and Resistance Mechanisms Using the Second-Generation ALK Inhibitor AP26113 in Human NPM-ALK–Positive Anaplastic Large Cell Lymphoma

“…S6) and lies in a hydrophobic cluster composed by F1098, F1271, and F1245. Mutations involving F1174 were recognized as activating in neuroblastoma and phenylalanine substitution with a leucine was found in an IMT patient that relapsed after Crizotinib treatment (24). Clones carrying cysteine, valine, or isoleucine in residue 1174 instead of phenylalanine were selected at AP26113 100 nmol/L and 200 nmol/L in a previous Ba/F3 screen (34).…”

“…However, despite great success, as expected from previous experience with tyrosine kinase inhibitors (TKI; refs. 21,22), the first cases of relapse due to the positive selection of mutant clones have already been detected in several Crizotinib-treated NSCLC and ALCL patients (20,(23)(24)(25)(26). To effectively overcome Crizotinib resistance, the development of second-generation ALK inhibitors is exponentially growing (27).…”

Therapeutic strategies to overcome crizotinib resistance in non-small cell lung cancers harboring the fusion oncogene EML4-ALK

“…EML4-ALK-positive NSCLC represents another tyrosine kinasedriven cancer that is highly responsive to TKI therapy. Recently, two studies have reported the identification of secondary resistance mutations within the ALK TK domain in patients who relapsed on crizotinib (23,33). One of the mutations identified was the gatekeeper L1196M substitution, analogous to T315I in ABL and T790M in EGFR (23).…”

The R1275Q Neuroblastoma Mutant and Certain ATP-competitive Inhibitors Stabilize Alternative Activation Loop Conformations of Anaplastic Lymphoma Kinase

“…As with the ALK fusion proteins, the neuroblastoma activating mutants are amenable to inhibition by small molecule inhibitors of the ALK kinase activity, although differential sensitivity has been observed depending on the particular inhibitor and mutant (33,35). Interestingly, the F1174L variant and the related F1174C variant have been independently identified in the clinic as a mutations conferring resistance to crizotinib treatment (36,37).…”