Abstract PO045: Inhibition of GAS6/AXL improves efficacy of HER2 inhibitor trastuzumab in uterine serous cancer
Authors: Joan Tankou, Michael Toboni, Hollie Noia, Alyssa Oplt, Daniel Wilke, Dineo Khabele, Lindsay Kuroki, Andrea Hagemann, Carolyn McCourt, Premal Thaker, David Mutch, Matthew Powell, Katherine Fuh
Journal: Clinical Cancer Research (2021)

Abstract Objectives: To determine whether inhibition of GAS6/AXL can improve efficacy to the humanized anti-HER2 monoclonal antibody trastuzumab by decreasing uterine serous cancer cell invasion and cell survival in vitro and tumor burden in vivo. Methods: We treated AXL-expressing and HER2-expressing uterine serous cancer cell lines (ARK1, ARK2) with trastuzumab, AVB-500, or trastuzumab plus AVB-500. Clonogenic cell survival assays were performed. Matrigel invasion assays were used to assess the invasive capacity of ARK2 cells after 24 hours of treatment with vehicle, AVB-500, trastuzumab, or trastuzumab plus AVB-500. A proximity ligation assay was used to examine colocalization of HER2/neu and AXL. Western blots were performed to assess the effects of the individual drugs and combination on phosphorylation of HER2/neu and AXL. An intraperitoneal tumor model with 10 million ARK1 cells was treated with vehicle, AVB-500, trastuzumab, or trastuzumab plus AVB-500 for 35 days. The number, size, and volume of tumor nodules were measured. GraphPad Prism was used for statistical analysis. Results: We found decreased cell survival by clonogenic assays in ARK1 and ARK2 cells treated with trastuzumab plus AVB-500 compared to trastuzumab alone (relative absorbance 0.155 nm vs 0.202 nm, P=0.015). There was significantly less invasion from treatment with trastuzumab plus AVB-500 than with trastuzumab alone (3.2 vs 17.8 invading tumor cells/hpf, P=0.0004). Western blot showed that cells treated with trastuzumab plus AVB-500 had reduced phospho-AXL compared to cells treated with AVB-500 or trastuzumab alone. Furthermore, the proximity ligation assay demonstrated co-localization of the AXL and HER2 receptors in both ARK1 and ARK2 cells indicating a physical closeness that could account for this co-regulation. We found that mice treated with trastuzumab plus AVB-500 developed significantly less tumor burden than mice treated with trastuzumab alone (0.03205 g vs 0.08316 g, P= 0.024), AVB-500 alone (0.0320 g vs 0.1638 g, P&amp;lt;0.0001), and vehicle alone (0.03205 g vs 0.1154 g, P= 0.0395). Conclusions: The GAS6/AXL inhibitor AVB-500 potentiated the effect of trastuzumab to decrease uterine serous cancer cell proliferation and invasion in vitro and tumor burden in vivo likely through AXL and HER2 complexes. Citation Format: Joan Tankou, Michael Toboni, Hollie Noia, Alyssa Oplt, Daniel Wilke, Dineo Khabele, Lindsay Kuroki, Andrea Hagemann, Carolyn McCourt, Premal Thaker, David Mutch, Matthew Powell, Katherine Fuh. Inhibition of GAS6/AXL improves efficacy of HER2 inhibitor trastuzumab in uterine serous cancer [abstract]. In: Proceedings of the AACR Virtual Special Conference: Endometrial Cancer: New Biology Driving Research and Treatment; 2020 Nov 9-10. Philadelphia (PA): AACR; Clin Cancer Res 2021;27(3_Suppl):Abstract nr PO045.