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

查看全文 作　　者：Yuanzhong [1]Wu;Liwen [1]Zhou;Zifeng [1]Wang;Xin [1]Wang;Ruhua [1]Zhang;Lisi [1]Zheng;Tiebang [1]Kang

机构地区：[1]Sun Yat-sen University Cancer Center,State Key Laboratory of Oncology in South China,Collaborative Innovation Center for Cancer Medicine,Guangzhou 510060,China

出　　处：《Cancer Biology & Medicine》索引2020年第17卷第3期,共13页

基　　金：This work was funded by the National Key Research and Development Program of China(Grant No.2016YFA0500304to T.K.);the Science and Technology Program of Guangzhou,(Grant Nos.202002020092 and 201607020038 to T.K.);the National Nature Science Foundation in China(NSFC)(Grant Nos.81772922 to Y.W.,81702890 to X.W.,81530081,31571395 to T.K.);the Guangdong Natural Science Foundation Team Project(Grant No.2014A030312015 to T.K.);the Natural Science Foundation of Guangdong Province(Grant No.2016A030310218 to W.Y.). 摘　　要：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. 关键词：OSTEOSARCOMA KINASE CRISPR-Cas9 library TRRAP PKMYT1 TP53RK

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

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