Targeting hypoxia downstream signaling protein, CAIX, for CAR T-cell therapy against glioblastoma

作者：Jing Cui, Qi Zhang, Qi Song, Herui Wang, Pauline Dmitriev, Mitchell Sun, Xiaoyu Cao, Yang Wang, Liemei Guo, Iris H. Indig, Jared S. Rosenblum, Chunxia Ji, Dongqing Cao, Kaiyong Yang, Mark R. Gilbert, Yu Yao, Zhuang Zhengping · 发表于：Neuro-Oncology · 年份：2019 · DOI：10.1093/neuonc/noz117 · 被引用次数：72 · 研究领域：CAR-T cell therapy research、Virus-based gene therapy research、Immune Cell Function and Interaction

BACKGROUND: Glioblastoma survival remains unchanged despite continuing therapeutic innovation. Herein, we aim to (i) develop chimeric antigen receptor (CAR) T cells with a specificity to a unique antigen, carbonic anhydrase IX (CAIX), which is expressed in the hypoxic microenvironment characteristic of glioblastoma, and (ii) demonstrate its efficacy with limited off-target effects. METHODS: First we demonstrated expression of CAIX in patient-derived glioblastoma samples and available databases. CAR T cells were generated against CAIX and efficacy was assessed in 4 glioblastoma cell lines and 2 glioblastoma stem cell lines. Cytotoxicity of anti-CAIX CAR T cells was assessed via interferon gamma, tumor necrosis factor alpha, and interleukin-2 levels when co-cultured with tumor cells. Finally, we assessed efficacy of direct intratumoral injection of the anti-CAIX CAR T cells on an in vivo xenograft mouse model using the U251 luciferase cell line. Tumor infiltrating lymphocyte analyses were performed. RESULTS: We confirm that CAIX is highly expressed in glioblastoma from patients. We demonstrate that CAIX is a suitable target for CAR T-cell therapy using anti-CAIX CAR T cells against glioblastoma in vitro and in vivo. In our mouse model, a 20% cure rate was observed without detectable systemic effects. CONCLUSIONS: By establishing the specificity of CAIX under hypoxic conditions in glioblastoma and highlighting its efficacy as a target for CAR T-cell therapy, our data suggest tha...