Generation and Functional Verification of Hypoxiasensitive Chimeric Antigen Receptor-T Cells

Xue, Ying1; Mao, Yunyu2; Liao, Qibin3,4; Zhao, Chen5; Zhang, Xiaoyan1,2,4; Xu, Jianqing1,2,4

2024-06-01

10.3791/66697

JOVE-JOURNAL OF VISUALIZED EXPERIMENTS   影响因子和分区

1940-087X

Extensive studies have proven the promise of chimeric antigen receptor T (CAR-T) cell therapy in treating hematological malignancies. However, treating solid tumors remains challenging, as exemplified by the safety concerns that arise when CAR-T cells attack normal cells expressing the target antigens. Researchers have explored various approaches to enhance the tumor selectivity of CAR-T cell therapy. One representative strategy along this line is the construction of hypoxia-sensitive CART cells, which are designed by fusing an oxygen-dependent degradation domain to the CAR moiety and are strategized to attain high CAR expression only in a hypoxic environment-the tumor microenvironment (TME). This paper presents a protocol for the generation of such CAR-T cells and their functional characterization, including methods to analyze the changes in CAR expression and killing capacity in response to different oxygen levels established by a mobile incubator chamber. The constructed CAR-T cells are anticipated to demonstrate CAR expression and cytotoxicity in an oxygen-sensitive manner, thus supporting their capability to distinguish between hypoxic TME and normoxic normal tissues for selective activation.

SCI

英语

其他

National Key Research and Development Program of China[2016YFC1303402] ; National Megaproject on Key Infectious Diseases["2017ZX10202102","2017ZX10304402-002-007"] ; General Program of Shanghai Municipal Health Commission[201740194]

Science & Technology - Other Topics

Multidisciplinary Sciences

WOS:001266777300022

JOURNAL OF VISUALIZED EXPERIMENTS

Web of Science

1.Fudan Univ, Inst Biomed Sci, Shanghai, Peoples R China
2.Fudan Univ, Clin Ctr Biotherapy Zhongshan Hosp, Clin Ctr Biotherapy, Shanghai, Peoples R China
3.Southern Univ Sci & Technol, Shenzhen Peoples Hosp 3, Shenzhen, Peoples R China
4.Fudan Univ, Shanghai Publ Hlth Clin Ctr, Sci Res Ctr, Shanghai, Peoples R China
5.Fudan Univ, Shanghai Publ Hlth Clin Ctr, Shanghai, Peoples R China

Xue, Ying,Mao, Yunyu,Liao, Qibin,et al. Generation and Functional Verification of Hypoxiasensitive Chimeric Antigen Receptor-T Cells[J]. JOVE-JOURNAL OF VISUALIZED EXPERIMENTS,2024(208).

Xue, Ying,Mao, Yunyu,Liao, Qibin,Zhao, Chen,Zhang, Xiaoyan,&Xu, Jianqing.(2024).Generation and Functional Verification of Hypoxiasensitive Chimeric Antigen Receptor-T Cells.JOVE-JOURNAL OF VISUALIZED EXPERIMENTS(208).

Xue, Ying,et al."Generation and Functional Verification of Hypoxiasensitive Chimeric Antigen Receptor-T Cells".JOVE-JOURNAL OF VISUALIZED EXPERIMENTS .208(2024).