Development of nanobodies targeting hepatocellular carcinoma and application of nanobody-based CAR-T technology

作者：Keming Lin, Baijin Xia, Xuemei Wang, Xin He, Mo Zhou, Yingtong Lin, Yidan Qiao, Rong Li, Qier Chen, Yuzhuang Li, Jinzhu Feng, Tao Chen, Cancan Chen, Xinyu Li, Hui Zhang, Lijuan Lu, Bingfeng Liu, Xu Zhang · 发表于：Journal of Translational Medicine · 年份：2024 · DOI：10.1186/s12967-024-05159-x · 被引用次数：15 · 研究领域：Monoclonal and Polyclonal Antibodies Research、CAR-T cell therapy research、Transgenic Plants and Applications

BACKGROUND: Chimeric antigen receptor T (CAR-T) cell therapy, as an emerging anti-tumor treatment, has garnered extensive attention in the study of targeted therapy of multiple tumor-associated antigens in hepatocellular carcinoma (HCC). However, the suppressive microenvironment and individual heterogeneity results in downregulation of these antigens in certain patients' cancer cells. Therefore, optimizing CAR-T cell therapy for HCC is imperative. METHODS: In this study, we administered FGFR4-ferritin (FGFR4-HPF) nanoparticles to the alpaca and constructed a phage library of nanobodies (Nbs) derived from alpaca, following which we screened for Nbs targeting FGFR4. Then, we conducted the functional validation of Nbs. Furthermore, we developed Nb-derived CAR-T cells and evaluated their anti-tumor ability against HCC through in vitro and in vivo validation. RESULTS: Our findings demonstrated that we successfully obtained high specificity and high affinity Nbs targeting FGFR4 after screening. And the specificity of Nbs targeting FGFR4 was markedly superior to their binding to other members of the FGFR family proteins. Furthermore, the Nb-derived CAR-T cells, targeting FGFR4, exhibited significantly enhanced anti-tumor efficacy in both experiments when in vitro and in vivo. CONCLUSIONS: In summary, the results of this study suggest that the CAR-T cells derived from high specificity and high affinity Nbs, targeting FGFR4, exhibited significantly enhanced anti-tumor efficacy in vitr...