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...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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China Anti-Cancer Association
2020-08-01
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| Series: | Cancer Biology & Medicine |
| Subjects: | |
| Online Access: | http://www.cancerbiomed.org/index.php/cocr/article/view/1683 |
| _version_ | 1828497015739252736 |
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| author | Yuanzhong Wu Liwen Zhou Zifeng Wang Xin Wang Ruhua Zhang Lisi Zheng Tiebang Kang |
| author_facet | Yuanzhong Wu Liwen Zhou Zifeng Wang Xin Wang Ruhua Zhang Lisi Zheng Tiebang Kang |
| author_sort | Yuanzhong Wu |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-12-11T12:40:03Z |
| format | Article |
| id | doaj.art-0b1b4c22ec874c668b87891023d2b8bf |
| institution | Directory Open Access Journal |
| issn | 2095-3941 |
| language | English |
| last_indexed | 2024-12-11T12:40:03Z |
| publishDate | 2020-08-01 |
| publisher | China Anti-Cancer Association |
| record_format | Article |
| series | Cancer Biology & Medicine |
| spelling | doaj.art-0b1b4c22ec874c668b87891023d2b8bf2022-12-22T01:06:59ZengChina Anti-Cancer AssociationCancer Biology & Medicine2095-39412020-08-0117378279410.20892/j.issn.2095-3941.2020.0162Systematic screening for potential therapeutic targets in osteosarcoma through a kinome-wide CRISPR-Cas9 libraryYuanzhong Wu0Liwen Zhou1Zifeng Wang2Xin Wang3Ruhua Zhang4Lisi Zheng5Tiebang Kang6Sun Yat-sen University Cancer Center, State Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine, Guangzhou 510060, ChinaSun Yat-sen University Cancer Center, State Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine, Guangzhou 510060, ChinaSun Yat-sen University Cancer Center, State Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine, Guangzhou 510060, ChinaSun Yat-sen University Cancer Center, State Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine, Guangzhou 510060, ChinaSun Yat-sen University Cancer Center, State Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine, Guangzhou 510060, ChinaSun Yat-sen University Cancer Center, State Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine, Guangzhou 510060, ChinaSun Yat-sen University Cancer Center, State Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine, Guangzhou 510060, ChinaObjective: 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.http://www.cancerbiomed.org/index.php/cocr/article/view/1683osteosarcomakinasecrispr-cas9 librarytrrappkmyt1tp53rk |
| spellingShingle | Yuanzhong Wu Liwen Zhou Zifeng Wang Xin Wang Ruhua Zhang Lisi Zheng Tiebang Kang Systematic screening for potential therapeutic targets in osteosarcoma through a kinome-wide CRISPR-Cas9 library Cancer Biology & Medicine osteosarcoma kinase crispr-cas9 library trrap pkmyt1 tp53rk |
| 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 | osteosarcoma kinase crispr-cas9 library trrap pkmyt1 tp53rk |
| url | http://www.cancerbiomed.org/index.php/cocr/article/view/1683 |
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