Abstract 5826: Comprehensive analysis with single-cell RNA sequence and spatial transcriptomics unravels distinctive expression profile of apoptotic-related genes in EGFR-mutated lung cancer
Authors: Izumi Motohiro , Fujii Masanori , Ho Vivian , Kobayashi Ikei S. , Kobayashi Susumu S. 
Published: March 2024

Despite an initial marked response to targeted therapy with tyrosine kinase inhibitors (TKIs) in EGFR-mutated lung cancer, the duration of response is limited due to the inevitable development of acquired resistance. Anti- and pro-apoptotic genes play an important role in the survival of tumor cells. Currently, variations in these gene expression levels are not well characterized due to the complex interplay between the inducement of expression changes by EGFR-TKIs and the presence of inter- and intra-tumor heterogeneity. Here, a comprehensive analysis of apoptosis-related genes, including BCL-2 family members and the IAP family members, was conducted via single-cell RNA sequence (scRNA-seq) and spatial transcriptomics. scRNA-seq captured changes in apoptosis-related gene expression after EGFR-TKI treatment using different EGFR-mutated lung cancer cell lines. Notably, BCL2L1 expression increased, whereas MCL1 expression decreased after EGFR-TKI treatment. Other anti- and pro-apoptotic genes did not display consistent trends in the cell lines used. Additionally, scRNA-seq was carried out on samples collected from patients with EGFR-mutated lung cancer after the development of resistance to EGFR-TKIs in order to validate the results obtained from cell line data and to compare gene expression levels between tumor and non-tumor cells. BCL2L1 exhibited high expression specifically in tumor cells; in contrast, MCL1 expression was lower in tumor cells compared to non-tumor cells. Moreover, BAD and XIAP were highly expressed in a tumor-specific manner. To further corroborate the scRNA-seq findings and delineate the spatial heterogeneity in tumors, spatial transcriptomics was performed using EGFR-driven lung cancer transgenic mouse specimens in three conditions: prior to treatment, during therapeutic response after short-term treatment, and following acquired resistance to EGFR-TKIs. Spatial transcriptomics data demonstrated that an increased BCL2L1 expression was observed in tumor tissues and that MCL1 expression was significantly higher in tissue adjacent to tumors than in tumors. Finally, we demonstrated that genetic ablation of BCL2L1 and pharmacological inhibition of BCL2L1 overcame or delayed resistance to EGFR-TKIs in vitro and in vivo. Overall, BCL2L1 is particularly important for tumor cell survival during the emergence of drug tolerance and the development of acquired resistance to EGFR-TKIs.

Citation Format: Motohiro Izumi, Masanori Fujii, Vivian Ho, Ikei S. Kobayashi, Susumu S. Kobayashi. Comprehensive analysis with single-cell RNA sequence and spatial transcriptomics unravels distinctive expression profile of apoptotic-related genes in EGFR-mutated lung 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 5826.