MET gene exon 14 deletion created using the CRISPR/Cas9 system enhances cellular growth and sensitivity to a MET inhibitor.

Abstract

BACKGROUND:

MET splice site mutations resulting in an exon 14 deletion have been reported to be present in about 3% of all lung adenocarcinomas. Patients with lung adenocarcinoma and a MET splice site mutation who have responded to MET inhibitors have been reported. The CRISPR/Cas9 system is a recently developed genome-engineering tool that can easily and rapidly cause small insertions or deletions.

MATERIALS AND METHODS:

We created an in vitro model for MET exon 14 deletion using the CRISPR/Cas9 system and the HEK293 cell line. The phenotype, which included MET inhibitor sensitivity, was then investigated in vitro. Additionally, MET splice site mutations were analyzed in several cancers included in The Cancer Genome Atlas (TCGA) dataset.

RESULTS:

An HEK293 cell line with a MET exon 14 deletion was easily and rapidly created; this cell line had a higher MET protein expression level, enhanced MET phosphorylation, and prolonged MET activation. In addition, a direct comparison of phenotypes using this system demonstrated enhanced cellular growth, colony formation, and MET inhibitor sensitivity. In the TCGA dataset, lung adenocarcinomas had the highest incidence of MET exon 14 deletions, while other cancers rarely carried such mutations. Approximately 10% of the lung adenocarcinoma samples without any of driver gene alterations carried the MET exon 14 deletion.

CONCLUSIONS:

These findings suggested that this system may be useful for experiments requiring the creation of specific mutations, and the present experimental findings encourage the development of MET-targeted therapy against lung cancer carrying the MET exon 14 deletion.

• Authors: Yosuke Togashi, Hiroshi Mizuuchi, Shuta Tomida, Masato Terashima, Hidetoshi Hayashi, Kazuto Nishio, Tetsuya Mitsudomi
• Journal: Lung cancer (Amsterdam, Netherlands) (Lung Cancer) Vol. 90 Issue 3 Pg. 590-7 (Dec 2015) ISSN: 1872-8332 [Electronic] Ireland
• PMID: 26547802 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

Chemical References

• Protein Kinase Inhibitors
• RNA Splice Sites
• RNA, Guide
• Proto-Oncogene Proteins c-met

Topics

• Base Sequence
• CRISPR-Cas Systems
• Cell Line, Tumor
• Cell Proliferation
• Computational Biology (methods)
• Databases, Genetic
• Drug Resistance, Neoplasm (genetics)
• Exons
• Gene Targeting
• HEK293 Cells
• Humans
• Lung Neoplasms (drug therapy, genetics)
• Molecular Sequence Data
• Mutation
• Protein Kinase Inhibitors (pharmacology)
• Proto-Oncogene Proteins c-met (chemistry, genetics)
• RNA Splice Sites
• RNA, Guide, Kinetoplastida
• Sequence Analysis, DNA
• Sequence Deletion