Kinase inhibitors have revolutionized cancer treatment but their use has led to drug resistance, prompting development of follow-on inhibitors for resistant tumors. Anaplastic lymphoma kinase (ALK) was discovered by Dr. Stephan Morris (Insight Genetics? collaborator on these studies). ALK fusion genes and activating point mutations cause several cancers, and multiple pharmaceutical firms are developing ALK inhibitors. One inhibitor ? PF-02341066 (crizotinib, Pfizer) ? has entered Phase III trials and will likely receive FDA approval in 2011. The Morris group has identified a large number of ALK kinase domain mutations that confer high-level PF-02341066 resistance, which has already been reported in patients. A clinical genetic test to identify inhibitor-resistance mutations is needed to guide the development and use of next-generation ALK inhibitors for patients resistant to 1st-generation therapy. The work proposed herein will develop a clinical diagnostic assay for ALK inhibitor-resistance mutations to address this unmet need. Objective 1. Development and application of a genetic diagnostic assay to detect all known ALK inhibitor-resistance mutations using an allele-specific PCR platform. Objective 2: Demonstration and validation of the ability of the test to identify ALK mutations from both cell lines with ALK inhibitor resistance and re-biopsied lung cancer specimens from patients who developed PF-02341066 resistance.
