Abstract 5708: Silence exportin 7 sensitizes esophageal squamous cell cancer to CDK4/6 inhibition through block the cell cycle coresectory

Abstract

Abstract Introduction: There is a lack of effective target therapies for esophageal squamous cell cancer. Our previous studies have demonstrated that CDK4/6 is a potency target for ESCC and combination with ERBB inhibitors is sufficient to achieve durable response. The aim of the current study is to prospectively map the landscape of sensitivity and resistance to CDK4/6 inhibition and develop novel combination therapies in ESCC. Experimental Procedures: We performed a genome-wide functional genetic screening and mini-pool designed CRISPR screen approaches to identify the modulators whose depletion sensitize CDK4/6 inhibition or confer resistance to the CDK4/6 inhibitor Palbociclib and elucidate new therapeutic strategies. Results: Our screen identified RB as the most enriched sgRNA, while guides targeting CCNE1 and CDK2 as the most depletion sgRNAs after CDK4/6 inhibition. These results validates our experimental approach as RB loss and CCNE1/CDK2 reactivation is the main mechanisms of CDK4/6 resistance. After gathering potential candidates from both screens, we found that genetically ablation of Exportin 7, a nuclear export factor, strongly sensitize CDK4/6 inhibition both in vitro and in vivo. RNAseq revealed a further repression of transcriptional activity of E2F and G2M by addition of XPO7depletion under CDK4/6 inhibitor treatment. To increase our insight in the molecular mechanism that could explain the synergy between XPO7 and CDK4/6, we further performed proteomics of nuclear and cytoplasm in ESCC lines with or without CDK4/6 inhibition. We found that silence XPO7 result in nuclear accumulation of mitosis-regulated genes such as CENPM, KIF4A and WEE1. Combined CDK4/6 inhibition with XPO7 depletion results in further downregulate of CENPM and KIF4A expression in nucleus, abrogating compensatory activation of cell cycle regulation and promote the cytostatic effect in ESCC. We further validated that CENPM and KIF4A depletion also sensitize CDK4/6 inhibition, and consequentially use WEE1 inhibitor after CDK4/6 inhibitor also effectively block the growth of ESCC lines. Conclusion: Overall, our comprehensive functional genetic screening approach revealed modulation of sensitive and resistance to the inhibition of CDK4/6 in ESCC. XPO7 was identified as a driver to change cell cycle landscape and merits as a therapeutic target combined with CDK4/6 inhibition in ESCC, which is beneficial to provide a novel treatment strategy. Citation Format: Zhong Wu, Lin Xu, Chuang Jiang, Adam Bass, Jin Zhou. Silence exportin 7 sensitizes esophageal squamous cell cancer to CDK4/6 inhibition through block the cell cycle coresectory [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 5708.