Original articles
Huan Deng, Songwei Tan, Xueqin Gao, Chenming Zou, Chenfeng Xu, Kun Tu, Qingle Song, Fengjuan Fan, Wei Huang, Zhiping Zhang. Cdk5 knocking out mediated by CRISPR-Cas9 genome editing for PD-L1 attenuation and enhanced antitumor immunity[J]. Acta Pharmaceutica Sinica B, 2020, 10(2): 358-373

Cdk5 knocking out mediated by CRISPR-Cas9 genome editing for PD-L1 attenuation and enhanced antitumor immunity
Huan Denga, Songwei Tana, Xueqin Gaoa, Chenming Zoua, Chenfeng Xua, Kun Tua, Qingle Songa, Fengjuan Fanb, Wei Huangc, Zhiping Zhanga,d,e
a School of Pharmacy, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China;
b Institute of Hematology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China;
c Department of Orthopedics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China;
d National Engineering Research Center for Nanomedicine, Huazhong University of Science and Technology, Wuhan 430030, China;
e Hubei Engineering Research Center for Novel Drug Delivery System, Huazhong University of Science and Technology, Wuhan 430030, China
Abstract:
Blocking the programmed death-ligand 1 (PD-L1) on tumor cells with monoclonal antibody therapy has emerged as powerful weapon in cancer immunotherapy. However, only a minority of patients presented immune responses in clinical trials. To develop an alternative treatment method based on immune checkpoint blockade, we designed a novel and efficient CRISPR-Cas9 genome editing system delivered by cationic copolymer aPBAE to downregulate PD-L1 expression on tumor cells via specifically knocking out Cyclin-dependent kinase 5 (Cdk5) gene in vivo. The expression of PD-L1 on tumor cells was significantly attenuated by knocking out Cdk5, leading to effective tumor growth inhibition in murine melanoma and lung metastasis suppression in triple-negative breast cancer. Importantly, we demonstrated that aPBAE/Cas9-Cdk5 treatment elicited strong T cell-mediated immune responses in tumor microenvironment that the population of CD8+ T cells was significantly increased while regulatory T cells (Tregs) was decreased. It may be the first case to exhibit direct in vivo PD-L1 downregulation via CRISPR-Cas9 genome editing technology for cancer therapy. It will provide promising strategy for preclinical antitumor treatment through the combination of nanotechnology and genome engineering.
Key words:    CRISPR-Cas9 genome editing system    Cyclin-dependent kinase 5 (Cdk5)    Programmed death-ligand 1 (PD-L1)    Antitumor immunity    Nanoparticles   
Received: 2019-04-22     Revised: 2019-06-29
DOI: 10.1016/j.apsb.2019.07.004
Funds: This work was supported by the National Natural Science Foundation of China (81872810, 81673374 and 81871473), Wuhan University of Science and Technology Plan for Applied Fundamental Research (2017060201010146, China), the Fundamental Research Funds for the Central Universities (2018KFYYXJJ019, 2019KFYRCPY049 and 2016YXMS138, China). We thank Weijun Feng, Ph.D, Professor, Fudan University, for professional advice.
Corresponding author: Zhiping Zhang     Email:zhipingzhang@hust.edu.cn
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Huan Deng
Songwei Tan
Xueqin Gao
Chenming Zou
Chenfeng Xu
Kun Tu
Qingle Song
Fengjuan Fan
Wei Huang
Zhiping Zhang

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