Genome-scale CRISPR screening identifies cell cycle and protein ubiquitination processes as druggable targets for erlotinib-resistant lung cancer

Jieun Lee, Ahyoung Choi, Sung Yup Cho, Yukyung Jun, Deukchae Na, Ahra Lee, Giyong Jang, Jee Young Kwon, Jaesang Kim, Sanghyuk Lee, Charles Lee

Research output: Contribution to journalArticlepeer-review

13 Scopus citations

Abstract

Erlotinib is highly effective in lung cancer patients with epidermal growth factor receptor (EGFR) mutations. However, despite initial favorable responses, most patients rapidly develop resistance to erlotinib soon after the initial treatment. This study aims to identify new genes and pathways associated with erlotinib resistance mechanisms in order to develop novel therapeutic strategies. Here, we induced knockout (KO) mutations in erlotinib-resistant human lung cancer cells (NCI-H820) using a genome-scale CRISPR-Cas9 sgRNA library to screen for genes involved in erlotinib susceptibility. The spectrum of sgRNAs incorporated among erlotinib-treated cells was substantially different to that of the untreated cells. Gene set analyses showed a significant depletion of ‘cell cycle process’ and ‘protein ubiquitination pathway’ genes among erlotinib-treated cells. Chemical inhibitors targeting genes in these two pathways, such as nutlin-3 and carfilzomib, increased cancer cell death when combined with erlotinib in both in vitro cell line and in vivo patient-derived xenograft experiments. Therefore, we propose that targeting cell cycle processes or protein ubiquitination pathways are promising treatment strategies for overcoming resistance to EGFR inhibitors in lung cancer.

Original languageEnglish
Pages (from-to)487-502
Number of pages16
JournalMolecular Oncology
Volume15
Issue number2
DOIs
StatePublished - Feb 2021

Bibliographical note

Funding Information:
The authors would like to thank Jinjoo Kang and Seoyeon Min for assisting with experiments and Jane Cha for the illustrations. C. Lee was a distinguished Ewha Womans University Professor supported in part by the Ewha Womans University Research grant of 2016–2019. Also, this study is supported in part by the operational funds from The First Affiliated Hospital of Xi'an Jiaotong University. This study was supported by grants from the Ministry of Science and ICT via National Research Foundation (NRF‐2015K1A4A3047851 and NRF‐2014M3C9A3065221) and Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (NRF‐2018R1A6A3A11044849). in vivo

Publisher Copyright:
© 2020 The Authors. Molecular Oncology published by John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies

Keywords

  • CRISPR/Cas9 screening
  • cell cycle process
  • erlotinib resistance
  • lung cancer
  • protein ubiquitination pathway

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