Real world approach for molecular analysis of acquired EGFR TKI resistance mechanisms in non-small cell lung carcinoma

Abstract Background With the approval of first line osimertinib treatment in stage IV epidermal growth factor receptor (EGFR) mutated non-small cell lung cancer (NSCLC), detection of resistance mechanisms will become increasingly important – and complex. Clear guidelines for analyses of these resistance mechanisms are currently lacking. Here, we provide our recommendations for optimal molecular diagnostics in the post-EGFR TKI resistance setting. Methods We compared Molecular work-up strategies from three hospitals of 161 first or second generation EGFR TKI treated cases and 159 osimertinib treated cases. Laboratories used combinations of DNA next generation sequencing (NGS), RNA NGS, in situ hybridization (ISH) and immunohistochemistry. Results Resistance mechanisms were identified in 72 first generation TKI cases (51%) and 85 osimertinib cases (57%). RNA NGS, when performed, revealed fusions or exon skipping events in 4% of early-TKI cases and 10% of osimertinib cases. Ten of the 30 MET and HER2 amplifications were exclusively detected by ISH or immunohistochemistry (IHC), and not detected by DNA NGS, mostly due to low tumor-cell percentage ( Discussion Our real world data support a method for molecular diagnostics, consisting of a parallel combination of DNA NGS, RNA NGS, MET ISH and either HER2 ISH or IHC. Combining RNA and DNA isolation into one-step limits dropout rates. In case of financial or tissue limitations, a sequential approach is justifiable, in which RNA NGS is only performed in case no resistance mechanisms are identified, Yet, this is suboptimal as – although rare – multiple acquired resistance mechanisms may occur.