Abstract 5192: EGFR inhibition enhances the cellular uptake and antitumor activity of the novel HER3 antibody drug conjugate U3-1402
Authors: Heidi M. Haikala, Jens Köhler, Timothy Lopez, Pinar Eser, Man Xu, Channing Yu, Yoshinobu Shiose, Yang Qiu, Prafulla Gokhale, Pasi A. Jänne
Journal: Cancer Research (2020)

Abstract Epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKI) have changed the treatment landscape for EGFR-mutant non-small cell lung cancers (NSCLC); however, most patients develop resistance over time. HER3 is a unique pseudokinase member of the ERBB family which functions through dimerization with other ERBB family members (EGFR and HER2) and has been difficult to target with conventional kinase inhibitor strategies. HER3 is frequently over expressed in EGFR-mutant NSCLC. U3-1402 is an antibody-drug conjugate (ADC) comprised of HER3-targeting antibody (patritumab) linked to a topoisomerase I inhibitor (DX-8951 derivative, or DXd). U3-1402 is currently in clinical development as a single agent in metastatic EGFR-mutant NSCLC (NCT03260491). We aimed to develop a preclinical strategy to enhance the efficacy of U3-1402. Pre-treatment with EGFR TKIs (gefitinib or osimertinib) increased HER3 membrane levels in six different EGFR-mutant cell lines by increasing both the amount of HER3 positive cells and the intensity of HER3 expression. Furthermore, using a proximity-ligation assay (PLA), we noted an increase in membrane EGFR:HER3 interaction following EGFR inhibitor treatment. HER3 protein levels and EGFR:HER3 interaction were similarly increased in four EGFR-mutant patient-derived xenograft (PDX) models following osimertinib treatment. HER3 expression peaked between 24 and 48 hours, and the increase in HER3 was regulated by both increased gene expression and in the level of protein stability. We further evaluated the biological consequences of increased HER3 expression on U3-1402 intake and activity. We used pH-sensitive pHrodo Red conjugate to label U3-1402 to monitor the intake of the ADC using Incucyte live cell imaging. In three EGFR-mutant cell lines, there was a 3- to 5-fold higher intake of U3-1402 over time in cell lines pre-treated with osimertinib, compared to cell lines without pre-treatment. Osimertinib pre-treatment also increased the uptake of U3-1402 in HER3-low expressing EGFR-mutant models. In contrast, osimertinib pre-treatment did not increase the uptake of a control IgG-ADC. In long-term in vitro cell growth assays, the combination of osimertinib and U3-1402 was superior to single agent U3-1402 in four EGFR-mutant cell lines. Preliminary in vivo studies demonstrated no significant toxicities as measured by mouse body weights and complete blood counts. In vivo experiments evaluating the antitumor efficacy of the combination of osimertinib and U3-1402 are currently underway. Our studies reveal that EGFR inhibitor treatment increased membrane expression of HER3 which was associated with enhanced internalization of U3-1402 in EGFR-mutant NSCLC. The combination of osimertinib and U3-1402 may be an effective treatment approach and should be evaluated in future clinical trials in patients with EGFR-mutant NSCLC. Citation Format: Heidi M. Haikala, Jens Köhler, Timothy Lopez, Pinar Eser, Man Xu, Channing Yu, Yoshinobu Shiose, Yang Qiu, Prafulla Gokhale, Pasi A. Jänne. EGFR inhibition enhances the cellular uptake and antitumor activity of the novel HER3 antibody drug conjugate U3-1402 [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 5192.