American Association for Cancer Research
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Supplementary Figures S1 - S14, Tables S1 - S5 from Epithelial-to-Mesenchymal Transition Defines Feedback Activation of Receptor Tyrosine Kinase Signaling Induced by MEK Inhibition in KRAS-Mutant Lung Cancer

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journal contribution
posted on 2023-04-03, 21:01 authored by Hidenori Kitai, Hiromichi Ebi, Shuta Tomida, Konstantinos V. Floros, Hiroshi Kotani, Yuta Adachi, Satoshi Oizumi, Masaharu Nishimura, Anthony C. Faber, Seiji Yano

Supplementary Figure S1. Feedback activation of ERK signaling and upregulation of AKT signaling following MEK inhibitor treatment in KRAS mutant lung cancers. Supplementary Figure S2. Scattered plot analysis showing inverse relationship between ERBB3 and Vimentin and positive correlation between FGFR1 and Vimentin. p < 0.01, both by linear regression analysis. Supplementary Figure S3. MEK inhibition promotes activation of FRS2 in mesenchymal-like KRAS mutant lung cancer cells. Supplementary Figure S4. FGFR inhibition has minimal effect on downstream signaling in mesenchymal-like KRAS mutant cancer cell lines. Supplementary Figure S5. FGF modestly induces trametinib resistance in mesenchymal-like KRAS mutant lung cancer cell lines. Supplementary Figure S6. Results of microarray analysis showing strong suppression of DUSP6 and SPRY4 mRNA levels following trametinib treatment. Supplementary Figure S7. FGFR1-FRS2 pathway is involved in feedback activation of MAPK signaling in an EMT induced NCI-H358 epithelial-like KRAS mutant cancer cells. Supplementary Figure S8. Combination of MEK inhibition with FGFR inhibition leads to further ERK suppression and apoptosis in mesenchymal-like KRAS mutant lung cancer cell lines. Supplementary Figure S9. Magnitude of apoptosis induced by FGFR and MEK inhibition is related to that which is induced by PI3K and MEK inhibition in mesenchymal-like KRAS mutant lung cancer cell lines. Supplementary Figure S10. Suppression of both MAPK and PI3K signal is not enough to induce cell death in cells resistant to MEK inhibitor with FGFR inhibitor. Supplementary Figure S11. Combination of trametinib with NVP-BGJ398 was tolerable in a mouse xenograft model. Supplementary Figure S12. No relationship between sensitivity to combination of FGFR inhibitor with trametinib and mutations in p53 and/or LKB1 in mesenchymal-like KRAS mutant cancers. Supplementary Figure S13. No relationship between EMT and subgroups of KRAS mutant lung adenocarcinoma defined by a previous study. Supplementary Figure S14. No relationship between sensitivity to combination of FGFR inhibitor with trametinib and amino acid substitutions in KRAS in mesenchymal-like KRAS mutant cancers. Supplementary Table S1. Differentially expressed genes following EMT in the NCI-H358 cell line. Supplementary Table S2. No significant alterations in the expression of FGF ligands and receptors after trametinib treatment in NCI-H1792 cells. Supplementary Table S3. Differentially expressed genes by microarray analysis between control and trametinib-treated NCI-H1792 cells. Supplementary Table S4. Mutational status of cell lines used in this study. Supplementary Table S5. Details of the antibodies.


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Ministry of Education, Culture, Sports, Science, and Technology of Japan


Kobayashi Foundation for Cancer Research




KRAS is frequently mutated in lung cancer. Whereas MAPK is a well-known effector pathway of KRAS, blocking this pathway with clinically available MAPK inhibitors is relatively ineffective. Here, we report that epithelial-to-mesenchymal transition rewires the expression of receptor tyrosine kinases, leading to differential feedback activation of the MAPK pathway following MEK inhibition. In epithelial-like KRAS-mutant lung cancers, this feedback was attributed to ERBB3-mediated activation of MEK and AKT. In contrast, in mesenchymal-like KRAS-mutant lung cancers, FGFR1 was dominantly expressed but suppressed by the negative regulator Sprouty proteins; MEK inhibition led to repression of SPRY4 and subsequent FGFR1-mediated reactivation of MEK and AKT. Therapeutically, the combination of a MEK inhibitor (MEKi) and an FGFR inhibitor (FGFRi) induced cell death in vitro and tumor regressions in vivo. These data establish the rationale and a therapeutic approach to treat mesenchymal-like KRAS-mutant lung cancers effectively with clinically available FGFR1 and MAPK inhibitors.Significance: Adaptive resistance to MEKi is driven by receptor tyrosine kinases specific to the differentiation state of the KRAS-mutant non–small cell lung cancer (NSCLC). In mesenchymal-like KRAS-mutant NSCLC, FGFR1 is highly expressed, and MEK inhibition relieves feedback suppression of FGFR1, resulting in reactivation of ERK; suppression of ERK by MEKi/FGFRi combination results in tumor shrinkage. Cancer Discov; 6(7); 754–69. ©2016 AACR.This article is highlighted in the In This Issue feature, p. 681

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