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Genome‐wide CRISPR/Cas9 screening for therapeutic targets in NSCLC carrying wild‐type TP53 and receptor tyrosine kinase genes

BACKGROUND: Targeted drugs have greatly improved the therapeutic outcome of non‐small cell lung cancer (NSCLC) patients compared with conventional chemotherapy, whereas about one‐third of patients are so far not suitable for targeted therapy due to lack of known driver oncogenes such as a mutated re...

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Autores principales: Wang, Qianqian, Li, Jun, Zhu, Jing, Mao, Jiaqi, Duan, Chao, Liang, Xiao, Zhu, Lingyun, Zhu, Mengyan, Zhang, Zhihong, Lin, Fan, Guo, Renhua
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9189421/
https://www.ncbi.nlm.nih.gov/pubmed/35692096
http://dx.doi.org/10.1002/ctm2.882
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author Wang, Qianqian
Li, Jun
Zhu, Jing
Mao, Jiaqi
Duan, Chao
Liang, Xiao
Zhu, Lingyun
Zhu, Mengyan
Zhang, Zhihong
Lin, Fan
Guo, Renhua
author_facet Wang, Qianqian
Li, Jun
Zhu, Jing
Mao, Jiaqi
Duan, Chao
Liang, Xiao
Zhu, Lingyun
Zhu, Mengyan
Zhang, Zhihong
Lin, Fan
Guo, Renhua
author_sort Wang, Qianqian
collection PubMed
description BACKGROUND: Targeted drugs have greatly improved the therapeutic outcome of non‐small cell lung cancer (NSCLC) patients compared with conventional chemotherapy, whereas about one‐third of patients are so far not suitable for targeted therapy due to lack of known driver oncogenes such as a mutated receptor tyrosine kinase (RTK) genes. In this study, we aimed to identify therapeutic targets for this subgroup of NSCLC patients. METHODS: We performed genome‐wide CRISPR/Cas9 screens in two NSCLC cell lines carrying wild‐type TP53 and receptor tyrosine kinase (wtTP53‐RTK) genes using a GeCKO v2.0 lentiviral library (containing 123411 sgRNAs and targeting 19050 genes). MAGeCKFlute was used to analyse and identify candidate genes. Genetic perturbation and pharmacological inhibition were used to validate the result in vitro and in vivo. RESULTS: The Genome‐wide CRISPR/Cas9 screening identified MDM2 as a potential therapeutic target for wtTP53‐RTK NSCLC. Genetic and pharmacological inhibition of MDM2 reduced cell proliferation and impaired tumour growth in the xenograft model, thus confirming the finding of the CRISPR/Cas9 screening. Moreover, treatment by a selective MDM2 inhibitor RG7388 triggered both cell cycle arrest and apoptosis in several NSCLC cell lines. Additionally, RG7388 and pemetrexed synergistically blocked the cell proliferation and growth of wtTP53‐RTK tumours but had limited effects for other genotypes. CONCLUSIONS: We identified MDM2 as an essential gene and a potential therapeutic target in wtTP53‐RTK NSCLC via a genome‐wide CRISPR/Cas9 screening. For this subgroup, treatment by RG7388 alone or by its combination with pemetrexed resulted in significant tumour inhibition.
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spelling pubmed-91894212022-06-16 Genome‐wide CRISPR/Cas9 screening for therapeutic targets in NSCLC carrying wild‐type TP53 and receptor tyrosine kinase genes Wang, Qianqian Li, Jun Zhu, Jing Mao, Jiaqi Duan, Chao Liang, Xiao Zhu, Lingyun Zhu, Mengyan Zhang, Zhihong Lin, Fan Guo, Renhua Clin Transl Med Research Articles BACKGROUND: Targeted drugs have greatly improved the therapeutic outcome of non‐small cell lung cancer (NSCLC) patients compared with conventional chemotherapy, whereas about one‐third of patients are so far not suitable for targeted therapy due to lack of known driver oncogenes such as a mutated receptor tyrosine kinase (RTK) genes. In this study, we aimed to identify therapeutic targets for this subgroup of NSCLC patients. METHODS: We performed genome‐wide CRISPR/Cas9 screens in two NSCLC cell lines carrying wild‐type TP53 and receptor tyrosine kinase (wtTP53‐RTK) genes using a GeCKO v2.0 lentiviral library (containing 123411 sgRNAs and targeting 19050 genes). MAGeCKFlute was used to analyse and identify candidate genes. Genetic perturbation and pharmacological inhibition were used to validate the result in vitro and in vivo. RESULTS: The Genome‐wide CRISPR/Cas9 screening identified MDM2 as a potential therapeutic target for wtTP53‐RTK NSCLC. Genetic and pharmacological inhibition of MDM2 reduced cell proliferation and impaired tumour growth in the xenograft model, thus confirming the finding of the CRISPR/Cas9 screening. Moreover, treatment by a selective MDM2 inhibitor RG7388 triggered both cell cycle arrest and apoptosis in several NSCLC cell lines. Additionally, RG7388 and pemetrexed synergistically blocked the cell proliferation and growth of wtTP53‐RTK tumours but had limited effects for other genotypes. CONCLUSIONS: We identified MDM2 as an essential gene and a potential therapeutic target in wtTP53‐RTK NSCLC via a genome‐wide CRISPR/Cas9 screening. For this subgroup, treatment by RG7388 alone or by its combination with pemetrexed resulted in significant tumour inhibition. John Wiley and Sons Inc. 2022-06-12 /pmc/articles/PMC9189421/ /pubmed/35692096 http://dx.doi.org/10.1002/ctm2.882 Text en © 2022 The Authors. Clinical and Translational Medicine published by John Wiley & Sons Australia, Ltd on behalf of Shanghai Institute of Clinical Bioinformatics. https://creativecommons.org/licenses/by/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Articles
Wang, Qianqian
Li, Jun
Zhu, Jing
Mao, Jiaqi
Duan, Chao
Liang, Xiao
Zhu, Lingyun
Zhu, Mengyan
Zhang, Zhihong
Lin, Fan
Guo, Renhua
Genome‐wide CRISPR/Cas9 screening for therapeutic targets in NSCLC carrying wild‐type TP53 and receptor tyrosine kinase genes
title Genome‐wide CRISPR/Cas9 screening for therapeutic targets in NSCLC carrying wild‐type TP53 and receptor tyrosine kinase genes
title_full Genome‐wide CRISPR/Cas9 screening for therapeutic targets in NSCLC carrying wild‐type TP53 and receptor tyrosine kinase genes
title_fullStr Genome‐wide CRISPR/Cas9 screening for therapeutic targets in NSCLC carrying wild‐type TP53 and receptor tyrosine kinase genes
title_full_unstemmed Genome‐wide CRISPR/Cas9 screening for therapeutic targets in NSCLC carrying wild‐type TP53 and receptor tyrosine kinase genes
title_short Genome‐wide CRISPR/Cas9 screening for therapeutic targets in NSCLC carrying wild‐type TP53 and receptor tyrosine kinase genes
title_sort genome‐wide crispr/cas9 screening for therapeutic targets in nsclc carrying wild‐type tp53 and receptor tyrosine kinase genes
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9189421/
https://www.ncbi.nlm.nih.gov/pubmed/35692096
http://dx.doi.org/10.1002/ctm2.882
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