Nucleofection with Plasmid DNA for CRISPR/Cas9-Mediated Inactivation of Programmed Cell Death Protein 1 in CD133-Specific CAR T Cells

doi:10.1089/hum.2017.234

	Nucleofection with Plasmid DNA for CRISPR/Cas9-Mediated Inactivation of Programmed Cell Death Protein 1 in CD133-Specific CAR T Cells
	Hu, Bian 1; Zou, Yan2 ; Zhang, Linlin 1; Tang, Jiaxing2 ; Niedermann, Gabriele 4,5; Firat, Elke 4,5; Huang, Xingxu3 ; Zhu, Xuekai2
	2019-04
发表期刊	HUMAN GENE THERAPY (IF:3.9[JCR-2023],5.8[5-Year])
ISSN	1043-0342
卷号	30 期号:4 页码:446-458
发表状态	已发表
DOI	10.1089/hum.2017.234
摘要	CRISPR/Cas9-mediated programmed cell death protein 1 (PD-1) disruption in chimeric antigen receptor (CAR) T cells could be an appealing choice to improve the therapeutic efficacy of CAR T cells in an immunosuppressive tumor microenvironment. In most of the reported cases, Cas9 was delivered into T cells by way of electroporation with RNA or protein. However, transient expression of Cas9 by transfection with a plasmid encoding its gene is apparently simpler, as it avoids the steps of in vitro transcription of DNA or protein production. This study tried nucleofection into human primary T cells of plasmids encoding both CRISPR/Cas9 for disrupting the PD-1 gene and the piggyBac transposon system for expressing CD133-specific CAR in one reaction. Based on drug selection, CD133-specific CAR T cells were obtained in which, on average, 91.5% of the PD-1 gene sites were disrupted, but almost no Cas9 gene expression was found in the final engineered CAR T cells. The PD-1-deficient CD133-specific CAR T cells showed similar levels of cytokine secretion and improved proliferation and cytotoxicity in vitro, and enhanced inhibition of tumor growth in an orthotopic mouse model of glioma, compared to conventional CD133-CAR T cells. The described method could be useful for the production of PD-1-deficient CAR T cells for cancer immunotherapy.
关键词	chimeric antigen receptor immunotherapy CRISPR Cas9 transposon PD-1 CD133
收录类别	SCI ; SCIE
语种	英语
WOS研究方向	Biotechnology & Applied Microbiology ; Genetics & Heredity ; Research & Experimental Medicine
WOS类目	Biotechnology & Applied Microbiology ; Genetics & Heredity ; Medicine, Research & Experimental
WOS记录号	WOS:000463602500006
出版者	MARY ANN LIEBERT, INC
WOS关键词	GLIOBLASTOMA STEM-CELLS ; SLEEPING-BEAUTY ; PD-1 ; GENERATION ; BLOCKADE
原始文献类型	Article
引用统计	正在获取...
文献类型	期刊论文
条目标识符	https://kms.shanghaitech.edu.cn/handle/2MSLDSTB/34250
专题	免疫化学研究所_PI研究组_朱学锴组 T细胞工程实验室生命科学与技术学院_PI研究组_黄行许组生命科学与技术学院_博士生
通讯作者	Huang, Xingxu; Zhu, Xuekai
作者单位	1.Nanjing Univ, Natl Resource Ctr Mutant Mice, Model Anim Res Ctr, MOE Key Lab Model Anim Dis Study, Nanjing, Jiangsu, Peoples R China 2.ShanghaiTech Univ, SIAIS, 393 Middle Huaxia Rd, Shanghai 201210, Peoples R China 3.ShanghaiTech Univ, SLST, 393 Middle Huaxia Rd, Shanghai 201210, Peoples R China 4.Univ Freiburg, Fac Med, Dept Radiat Oncol, Freiburg, Germany 5.German Canc Res Ctr, German Canc Consortium DKTK Partner Site Freiburg, Heidelberg, Germany
通讯作者单位	上海科技大学
推荐引用方式 GB/T 7714	Hu, Bian,Zou, Yan,Zhang, Linlin,et al. Nucleofection with Plasmid DNA for CRISPR/Cas9-Mediated Inactivation of Programmed Cell Death Protein 1 in CD133-Specific CAR T Cells[J]. HUMAN GENE THERAPY,2019,30(4):446-458.
APA	Hu, Bian.,Zou, Yan.,Zhang, Linlin.,Tang, Jiaxing.,Niedermann, Gabriele.,...&Zhu, Xuekai.(2019).Nucleofection with Plasmid DNA for CRISPR/Cas9-Mediated Inactivation of Programmed Cell Death Protein 1 in CD133-Specific CAR T Cells.HUMAN GENE THERAPY,30(4),446-458.
MLA	Hu, Bian,et al."Nucleofection with Plasmid DNA for CRISPR/Cas9-Mediated Inactivation of Programmed Cell Death Protein 1 in CD133-Specific CAR T Cells".HUMAN GENE THERAPY 30.4(2019):446-458.