Abstract 6645: Anti-EGF antibodies significantly improve the activity of RET, BRAF, MEK and PI3K kinase inhibitors in preclinical models

Abstract

Abstract Background: We have recently shown that antibodies generated by vaccination (anti-EGF VacAbs) potentiate the effects of tyrosine kinase inhibitors (TKIs) in epidermal growth factor receptor mutant (EGFR-mut) cell lines (1) and a Phase I/II clinical trial of an anti-EGF vaccine in combination with afatinib has been initiated. In this study we aimed to determine the efficacy of anti-EGF VacAbs to improve the antitumor activity of RET, BRAF, MEK and PI3K inhibitors in non-small cell lung cancer (NSCLC) and colorectal cancer (CRC) cell lines. Methods: Cell lines with RET translocations (LC-2/ad) and BRAF, KRAS and PIK3CA mutations (HT29, DLD1, LS174T and H508) were used. Anti-EGF VacAbs were obtained by immunizing rabbits with recombinant human EGF. Cell lines were treated with anti-EGF VacAbs alone and in combination with RET, BRAF, MEK and PI3K inhibitors. Cell viability was determined by MTT, cell cycle was analyzed by flow cytometry, changes of total and phosphorylated proteins by Western blot and emergence of resistance by direct microscopic examination in low density cultures. Results: Anti-EGF VacAbs suppressed EGF-induced cell proliferation and blocked EGFR transduction signaling pathway in all cell lines tested. In combination, the anti-EGF VacAbs significantly enhanced the antitumor activity of BLU667 in LC-2/ad, trametinib and encorafenib in HT29, trametinib in DLD1 and LS174T and trametinib, taselisib, alpelisib and copanlisib in H508 cells. In these cell lines, anti-EGF VacAbs in combination with kinase inhibitors suppressed EGFR, Akt and Erk 1/2 phosphorylation. Cell cycle experiments revealed that anti-EGF VacAbs significantly increased the antiproliferative effects of the kinase inhibitors, measured as the percentage of cells in G2+M. Finally, the addition of the anti-EGF VacAbs to the culture medium significantly delayed the emergence resistant clones to trametinib in DLD1 cells. Conclusions: Anti-EGF VacAbs potentiate the antitumor effects of RET, MEK, BRAF and PI3K inhibitors in tumor cell lines. Our data provide a rationale for clinical trials testing the combination of anti-EGF VacAbs with kinase inhibitors in RET-translocated, KRAS, BRAF and PIK3CA mutant NSCLC and CRC patients.(1)“Anti-Epidermal Growth Factor Vaccine Antibodies Enhance the Efficacy of Tyrosine Kinase Inhibitors and Delay the Emergence of Resistance in EGFR Mutant Lung Cancer Cells” Codony-Servat J, García-Roman S, Molina-Vila MA, et al. J Thorac Oncol. 2018. Citation Format: Silvia Garcia Roman, Jordi Codony Servat, Miguel Angel Molina Vila, Jordi Bertran Alamillo, Monica Garzon, Alejandro Martínez Bueno, Santiago Viteri, María González Cao, Andrés Aguilar, Juan José García, Erik d'Hondt, Rafael Rosell. Anti-EGF antibodies significantly improve the activity of RET, BRAF, MEK and PI3K kinase inhibitors in preclinical models [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 6645.