The genomic landscape of sensitivity to arsenic trioxide uncovered by genome-wide CRISPR-Cas9 screening

Jun-Zhu Chen; Li-Na Wang; Xue-Qun Luo; Yan-Lai Tang

doi:10.3389/fonc.2023.1178686

The genomic landscape of sensitivity to arsenic trioxide uncovered by genome-wide CRISPR-Cas9 screening

Front Oncol. 2023 May 12:13:1178686. doi: 10.3389/fonc.2023.1178686. eCollection 2023.

Authors

Jun-Zhu Chen¹, Li-Na Wang¹, Xue-Qun Luo¹, Yan-Lai Tang¹

Affiliation

¹ Department of Pediatrics, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong, China.

Abstract

Introduction: Arsenic trioxide (ATO) is a promising anticancer drug for hematological malignancy. Given the dramatic efficacy of acute promyelocytic leukemia (APL), ATO has been utilized in other types of cancers, including solid tumors. Unfortunately, the results were not comparable with the effects on APL, and the resistance mechanism has not been clarified yet. This study intends to identify relevant genes and pathways affecting ATO drug sensitivity through genome-wide CRISPR-Cas9 knockdown screening to provide a panoramic view for further study of ATO targets and improved clinical outcomes.

Methods: A genome-wide CRISPR-Cas9 knockdown screening system was constructed for ATO screening. The screening results were processed with MAGeCK, and the results were subjected to pathway enrichment analysis using WebGestalt and KOBAS. We also performed protein-protein interaction (PPI) network analysis using String and Cytoscape, followed by expression profiling and survival curve analysis of critical genes. Virtual screening was used to recognize drugs that may interact with the hub gene.

Results: We applied enrichment analysis and identified vital ATO-related pathways such as metabolism, chemokines and cytokines production and signaling, and immune system responses. In addition, we identified KEAP1 as the top gene relating to ATO resistance. We found that KEAP1 expression was higher in the pan-cancer, including ALL, than in normal tissue. Patients with acute myeloid leukemia (AML) with higher KEAP1 expression had worse overall survival (OS). A virtual screen showed that etoposide and eltrombopag could bind to KEAP1 and potentially interact with ATO.

Discussion: ATO is a multi-target anticancer drug, and the key pathways regulating its sensitivity include oxidative stress, metabolism, chemokines and cytokines, and the immune system. KEAP1 is the most critical gene regulating ATO drug sensitivity, which is related to AML prognosis and may bind to some clinical drugs leading to an interaction with ATO. These integrated results provided new insights into the pharmacological mechanism of ATO and potentiate for further applications in cancer treatments.

Keywords: CRISPR-Cas9 screening; KEAP1; arsenic trioxide; leukemia; virtual screening.

Grants and funding

This work was funded by the Science and Technology Planning Project of Guangdong Province, China (Grant No.2021A0505060004), the Basic and Applied Basic Research Project of Guangdong Province, China (Grant No. 2021A1515111169), the Youth Talent Promotion Project of Guangzhou Association for Science and Technology, China (Grant No.X20210201015), the Science and Technology Planning Project of Guangzhou, China (Grant No. 202102020156), and the Grant Award from the Terry Fox Foundation, Canada.