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Systematic screening for potential therapeutic targets in osteosarcoma through a kinome-wide CRISPR-Cas9 library

Objective: Osteosarcoma is the most common primary malignant bone tumor. However, the survival of patients with osteosarcoma has remained unchanged during the past 30 years, owing to a lack of efficient therapeutic targets. Methods: We constructed a kinome-targeting CRISPR-Cas9 library containing 50...

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Detalles Bibliográficos
Autores principales: Wu, Yuanzhong, Zhou, Liwen, Wang, Zifeng, Wang, Xin, Zhang, Ruhua, Zheng, Lisi, Kang, Tiebang
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Compuscript 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7476084/
https://www.ncbi.nlm.nih.gov/pubmed/32944406
http://dx.doi.org/10.20892/j.issn.2095-3941.2020.0162
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author Wu, Yuanzhong
Zhou, Liwen
Wang, Zifeng
Wang, Xin
Zhang, Ruhua
Zheng, Lisi
Kang, Tiebang
author_facet Wu, Yuanzhong
Zhou, Liwen
Wang, Zifeng
Wang, Xin
Zhang, Ruhua
Zheng, Lisi
Kang, Tiebang
author_sort Wu, Yuanzhong
collection PubMed
description Objective: Osteosarcoma is the most common primary malignant bone tumor. However, the survival of patients with osteosarcoma has remained unchanged during the past 30 years, owing to a lack of efficient therapeutic targets. Methods: We constructed a kinome-targeting CRISPR-Cas9 library containing 507 kinases and 100 nontargeting controls and screened the potential kinase targets in osteosarcoma. The CRISPR screening sequencing data were analyzed with the Model-based Analysis of Genome-wide CRISPR/Cas9 Knockout (MAGeCK) Python package. The functional data were applied in the 143B cell line through lenti-CRISPR-mediated gene knockout. The clinical significance of kinases in the survival of patients with osteosarcoma was analyzed in the R2: Genomics Analysis and Visualization Platform. Results: We identified 53 potential kinase targets in osteosarcoma. Among these targets, we analyzed 3 kinases, TRRAP, PKMYT1, and TP53RK, to validate their oncogenic functions in osteosarcoma. PKMYT1 and TP53RK showed higher expression in osteosarcoma than in normal bone tissue, whereas TRRAP showed no significant difference. High expression of all 3 kinases was associated with relatively poor prognosis in patients with osteosarcoma. Conclusions: Our results not only offer potential therapeutic kinase targets in osteosarcoma but also provide a paradigm for functional genetic screening by using a CRISPR-Cas9 library, including target design, library construction, screening workflow, data analysis, and functional validation. This method may also be useful in potentially accelerating drug discovery for other cancer types.
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spelling pubmed-74760842020-09-16 Systematic screening for potential therapeutic targets in osteosarcoma through a kinome-wide CRISPR-Cas9 library Wu, Yuanzhong Zhou, Liwen Wang, Zifeng Wang, Xin Zhang, Ruhua Zheng, Lisi Kang, Tiebang Cancer Biol Med Original Article Objective: Osteosarcoma is the most common primary malignant bone tumor. However, the survival of patients with osteosarcoma has remained unchanged during the past 30 years, owing to a lack of efficient therapeutic targets. Methods: We constructed a kinome-targeting CRISPR-Cas9 library containing 507 kinases and 100 nontargeting controls and screened the potential kinase targets in osteosarcoma. The CRISPR screening sequencing data were analyzed with the Model-based Analysis of Genome-wide CRISPR/Cas9 Knockout (MAGeCK) Python package. The functional data were applied in the 143B cell line through lenti-CRISPR-mediated gene knockout. The clinical significance of kinases in the survival of patients with osteosarcoma was analyzed in the R2: Genomics Analysis and Visualization Platform. Results: We identified 53 potential kinase targets in osteosarcoma. Among these targets, we analyzed 3 kinases, TRRAP, PKMYT1, and TP53RK, to validate their oncogenic functions in osteosarcoma. PKMYT1 and TP53RK showed higher expression in osteosarcoma than in normal bone tissue, whereas TRRAP showed no significant difference. High expression of all 3 kinases was associated with relatively poor prognosis in patients with osteosarcoma. Conclusions: Our results not only offer potential therapeutic kinase targets in osteosarcoma but also provide a paradigm for functional genetic screening by using a CRISPR-Cas9 library, including target design, library construction, screening workflow, data analysis, and functional validation. This method may also be useful in potentially accelerating drug discovery for other cancer types. Compuscript 2020-08-15 2020-08-15 /pmc/articles/PMC7476084/ /pubmed/32944406 http://dx.doi.org/10.20892/j.issn.2095-3941.2020.0162 Text en Copyright: © 2020, Cancer Biology & Medicine http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Original Article
Wu, Yuanzhong
Zhou, Liwen
Wang, Zifeng
Wang, Xin
Zhang, Ruhua
Zheng, Lisi
Kang, Tiebang
Systematic screening for potential therapeutic targets in osteosarcoma through a kinome-wide CRISPR-Cas9 library
title Systematic screening for potential therapeutic targets in osteosarcoma through a kinome-wide CRISPR-Cas9 library
title_full Systematic screening for potential therapeutic targets in osteosarcoma through a kinome-wide CRISPR-Cas9 library
title_fullStr Systematic screening for potential therapeutic targets in osteosarcoma through a kinome-wide CRISPR-Cas9 library
title_full_unstemmed Systematic screening for potential therapeutic targets in osteosarcoma through a kinome-wide CRISPR-Cas9 library
title_short Systematic screening for potential therapeutic targets in osteosarcoma through a kinome-wide CRISPR-Cas9 library
title_sort systematic screening for potential therapeutic targets in osteosarcoma through a kinome-wide crispr-cas9 library
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7476084/
https://www.ncbi.nlm.nih.gov/pubmed/32944406
http://dx.doi.org/10.20892/j.issn.2095-3941.2020.0162
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