Abstract 4696: ERBB2 signaling and inhibition in bladder cancer

Abstract

Abstract The role of human epidermal growth factor receptor-2 (HER2) protein and the inhibition of its oncogenic signaling pathway, ERBB2, is well defined and found to be beneficial in combination with standard chemotherapies in breast and certain gastro-intestinal cancers. Targeting the ERBB2 pathway has had unclear or inconsistent results in early trials in bladder cancer (BLCA) and the efficacy of its combination with cisplatin, a standard BLCA chemotherapy, is not known. In this study, we identify the transcriptomic oncogenic signatures that correlate with ERBB2 signaling in BLCA, explore how HER2 expression in BLCA cell lines correlates with treatment response in vitro, and evaluate the potential of ERBB2 inhibition in combination with cisplatin for BLCA. We analyzed the tumor transcriptome from the Cancer Genome Atlas (TCGA) study of BLCA with single sample Gene Set Enrichment Analyses (ssGSEA) for Hallmark and Oncogenic gene sets. We performed multiple Pearson correlation analyses of HER2 protein levels and ERBB2 signaling with other signaling pathways in the TCGA BLCA tumors and adjusted the significance for multiple comparisons using the Benjamini-Hochberg method. Immunoblotting was used to assess HER2 protein levels across a panel of bladder cancer cell lines. We then assessed cell viability and colony formation upon exposure to the specific ERBB2 inhibitor, tucatinib. Synergy between cisplatin and tucatinib in BLCA cell lines was evaluated using the Chou-Talalay method. In the TCGA BLCA dataset, HER2 protein positively correlates with ERBB2 signaling with a Pearson correlation coefficient of 0.45 and an adjusted p-value of 1.73E-17. ERBB2 signaling has multiple significant positive and negative correlations. ERBB2 signaling negatively correlates with the DNA repair gene set with a Pearson coefficient of -0.5 and an adjusted p-value of 1.40E-22. HER2 protein expression varies greatly between bladder cancer cell lines: HT-1197 and UMUC3 have lower HER2 expression, with HT-1376, T24, RT4, 5637, and SW780 showing higher HER2 expression. HT-1197 and RT4 had the lowest IC50s for tucatinib and showed a strong response to low doses of tucatinib in the colony formation assay. Cisplatin and tucatinib have synergetic growth inhibition (combination index <1) at low concentration levels in the HT-1197 and SW780 cell lines. Multiple oncogenic pathways coexist with ERBB2 signaling in BLCA. The varied response to specific ERBB2 inhibition and the lack of correlation of the response with HER2 protein level in cell lines alludes to alternative oncogenic pathways. A strong negative correlation of ERBB2 signaling with the DNA response pathway in the TCGA BLCA dataset, combined with preclinical evidence of synergy between tucatinib and cisplatin, indicates a potential for development of the combination as a clinical treatment strategy and warrants further study. Citation Format: Constance Caparas-Spaugy, Chris Stubben, Ken Boucher, Bingjian Feng, Aaron Atkinson, David Nix, Bradley Cairns, Sumati Gupta. ERBB2 signaling and inhibition in bladder cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4696.