miR-6077 promotes cisplatin/pemetrexed resistance in lung adenocarcinoma via CDKN1A/cell cycle arrest and KEAP1/ferroptosis pathways

Guoshu Bi; Jiaqi Liang; Mengnan Zhao; Huan Zhang; Xing Jin; Tao Lu; Yuansheng Zheng; Yunyi Bian; Zhencong Chen; Yiwei Huang; Valeria Besskaya; Cheng Zhan; Qun Wang; Lijie Tan

doi:10.1016/j.omtn.2022.03.020

miR-6077 promotes cisplatin/pemetrexed resistance in lung adenocarcinoma via CDKN1A/cell cycle arrest and KEAP1/ferroptosis pathways

Mol Ther Nucleic Acids. 2022 Mar 28:28:366-386. doi: 10.1016/j.omtn.2022.03.020. eCollection 2022 Jun 14.

Authors

Guoshu Bi^{1

2}, Jiaqi Liang^{1

2}, Mengnan Zhao^{1

2}, Huan Zhang^{1

2}, Xing Jin^{1

2}, Tao Lu^{1

2}, Yuansheng Zheng^{1

2}, Yunyi Bian^{1

2}, Zhencong Chen^{1

2}, Yiwei Huang^{1

2}, Valeria Besskaya^{1

2}, Cheng Zhan^{1

2}, Qun Wang^{1

2}, Lijie Tan^{1

2}

Affiliations

¹ Department of Thoracic Surgery, Zhongshan Hospital, Fudan University, No. 180 Fenglin Rd, Xuhui District, Shanghai 200032, China.
² Cancer Center, Zhongshan Hospital, Fudan University, Shanghai 200032, China.

Abstract

Lung adenocarcinoma (LUAD) is one of the most common malignancies worldwide. Combination chemotherapy with cisplatin (CDDP) plus pemetrexed (PEM) remains the predominant therapeutic regimen; however, chemoresistance greatly limits its curative potential. Here, through CRISPR-Cas9 screening, we identified miR-6077 as a key driver of CDDP/PEM resistance in LUAD. Functional experiments verified that ectopic overexpression of miR-6077 desensitized LUAD cells to CDDP/PEM in both cell lines and patient-derived xenograft models. Through RNA sequencing in cells and single-cell sequencing of samples from patients with CDDP/PEM treatments, we observed CDDP/PEM-induced upregulation of CDKN1A and KEAP1, which in turn activated cell-cycle arrest and ferroptosis, respectively, thus leading to cell death. Through miRNA pull-down, we identified and validated that miR-6077 targets CDKN1A and KEAP1. Furthermore, we demonstrated that miR-6077 protects LUAD cells from cell death induced by CDDP/PEM via CDKN1A-CDK1-mediated cell-cycle arrest and KEAP1-NRF2-SLC7A11/NQO1-mediated ferroptosis, thus resulting in chemoresistance in multiple LUAD cells both in vitro and in vivo. Moreover, we found that GMDS-AS1 and LINC01128 sensitized LUAD cells to CDDP/PEM by sponging miR-6077. Collectively, these results imply the critical role of miR-6077 in LUAD's sensitivity to CDDP/PEM, thus providing a novel therapeutic strategy for overcoming chemoresistance in clinical practice.

Keywords: CRISPR-Cas9; MT: Non-coding RNAs; RNA-seq; cell-cycle arrest; chemoresistance; cisplatin; ferroptosis; homology-directed repair; lncRNA; miRNA; pemetrexed.