Generation of hypoxia-sensing chimeric antigen receptor T cells

Abstract

Summary: Exploiting hypoxia in solid malignancies to restrict expression of chimeric antigen receptors (CARs) on engineered T cells to the tumor microenvironment overcomes the risk of on-target off-tumor toxicity and minimizes tonic signaling, which promotes CAR T cell exhaustion. This protocol summarizes the synthetic biology underlying the development of a stringent oxygen-sensitive CAR for in vitro and in vivo preclinical characterization.For complete details on the use and execution of this protocol, please refer to Kosti et al. (2021).

Keywords

• Cell culture
• Cell isolation
• Flow cytometry/mass cytometry
• Cell-based assays
• Cell separation/fractionation
• Cancer