CRISPR/Cas9 screens unravel miR-3689a-3p regulating sorafenib resistance in hepatocellular carcinoma via suppressing CCS/SOD1-dependent mitochondrial oxidative stress

Yuanjun Lu, Yau Tuen Chan, Junyu Wu, Zixin Feng, Hongchao Yuan, Qiucheng Li, Tingyuan Xing, Lin Xu, Cheng Zhang, Hor-Yue Tan, Terence Kin-Wah Lee, Yibin Feng, Ning Wang*

*Corresponding author for this work

Research output: Contribution to journalJournal articlepeer-review

Abstract

Aims: Therapeutic outcome of sorafenib in hepatocellular carcinoma (HCC) is undermined by the development of drug resistance. This study aimed to identify the critical microRNA (miRNA) which is responsible for sorafenib resistance at the genomic level. 

Methods: CRISPR/Cas9 screen followed by gain- and loss-of-function assays both in vitro and in vivo were applied to identify the role of miR-3689a-3p in mediating sorafenib response in HCC. The upstream and downstream molecules of miR-3689a-3p and their mechanism of action were investigated. 

Results: CRISPR/Cas9 screening identified miR-3689a-3p was the most up-regulated miRNA in sorafenib sensitive HCC. Knockdown of miR-3689a-3p significantly increased sorafenib resistance, while its overexpression sensitized HCC response to sorafenib treatment. Proteomic analysis revealed that the effect of miR-3689a-3p was related to the copper-dependent mitochondrial superoxide dismutase type 1 (SOD1) activity. Mechanistically, miR-3689a-3p targeted the 3′UTR of the intracellular copper chaperone for superoxide dismutase (CCS) and suppressed its expression. As a result, miR-3689a-3p disrupted the intracellular copper trafficking and reduced SOD1-mediated scavenge of mitochondrial oxidative stress that eventually caused HCC cell death in response to sorafenib treatment. CCS overexpression blunted sorafenib response in HCC. Clinically, miR-3689a-3p was down-regulated in HCC and predicted favorable prognosis for HCC patients. 

Conclusion: Our findings provide comprehensive evidence for miR-3689a-3p as a positive regulator and potential druggable target for improving sorafenib treatment in HCC.

Original languageEnglish
Article number101015
JournalDrug Resistance Updates
Volume71
DOIs
Publication statusPublished - Nov 2023

Scopus Subject Areas

  • Oncology
  • Pharmacology
  • Cancer Research
  • Infectious Diseases
  • Pharmacology (medical)

User-Defined Keywords

  • In vivo CRISPR/Cas9 screen
  • Sorafenib
  • MiR-3689a-3p
  • Drug resistance of hepatocellular carcinoma
  • CCS

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