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              <p>Mutations affecting exon 14 splice sites of the gene encoding the MET receptor have been recently revealed in non-small cell lung cancer (NSCLC). These mutations induce MET exon 14 skipping (METex14), leading to receptor activation through deletion of a regulatory domain. Importantly, these mutations represent a promising therapeutic opportunity since MET tyrosine kinase inhibitors (TKI) are available. Nevertheless, these mutations raise several scientific and clinical questions. (i) Their functional consequences are still poorly understood,(ii) these mutations are highly heterogeneous which makes them difficult to detect, and (iii) efficacy of MET-TKI seems limited by largely unknown resistances. To address these issues, we first created by genome editing a panel of pulmonary cells expressing either METex14 or MET receptor mutated in each known active site of the regulatory domain. Comparison of signalling pathways, transcriptional landscapes and cellular responses revealed that METex14 activation is recapitulated by mutation of the CBL binding site involved in MET internalization, but provide also an unexpected resistance to apoptosis through abrogating its caspase cleavage. Second, in order to detect METex14 mutations in clinical routine practice, we developed an optimized targeted next generation sequencing panel covering the METex14 in addition to the usual targets. This panel revealed METex14 alterations in 2.2% NSCLC patients and presence of various concurrent alterations. Third, by further characterization of the concurrent alterations, we found high rate of PI3K pathway alterations in METex14 patients. In addition, MET-TKI treatment in 3 patients harboring these alterations had shown progressive disease, suggesting their involvement in resistance. Using a patient-derived cell line with primary resistance and cell lines in which MET or PI3K alterations were inserted, we confirmed involvement of PI3K activation in the resistance process, which was overcome with PI3K inhibitor. Overall, our integrated study reveals that METex14 mutations induce an original activation involving cooperation between regulatory mechanisms, but offering sensitivity to MET-TKI. Therefore, these mutations, now detectable in routine practice, are druggable by MET-TKI providing a novel therapeutic line for NSCLC, but have to face to innate resistance including PI3K alterations.</p>
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