@Article{or.2025.065394,
AUTHOR = {Tian-Tian Li, Ming-Yao Meng, Zheng Yu, Yang-Fan Guo, Yi-Yi Zhao, Hui Gao, Li-Li Yang, Li-Rong Yang, Meng-Yuan Chu, Shan He, Yuan Liu, Xiao-Dan Wang, Wen-Ju Wang, Zong-Liu Hou, Li-Wei Liao, Lin Li},
TITLE = {Investigation on the Anti-Cancer Effects of HER2-Targeted CAR-T Cells Engineered Using the <i>PiggyBac</i> Transposon System},
JOURNAL = {Oncology Research},
VOLUME = {33},
YEAR = {2025},
NUMBER = {11},
PAGES = {3447--3467},
URL = {http://www.techscience.com/or/v33n11/64062},
ISSN = {1555-3906},
ABSTRACT = { <b>Background:</b> Chimeric antigen receptor T (CAR-T) cell therapies have demonstrated significant clinical efficacy in hematological malignancies. However, their application to solid tumors remains substantially limited by multiple challenges, including the risk of off-target effects. Hence, optimizing CAR-T cells for stronger antigen binding is essential. <b>Methods:</b> In this study, we employed a classical anti-human endothelial growth factor receptor 2 (HER2) single-chain variable fragment (scFv) derived from trastuzumab, alongside an anti-HER2-13 scFv identified from a combinatorial cellular CAR library, for the construction of a third-generation CAR-T cell. Meanwhile, the phenotypes and both <i>in vitro</i> and <i>in vivo</i> functions of CAR-T cells transduced with the two scFvs via <i>PiggyBac</i> transposon-mediated gene transfer were compared. <b>Results:</b> The optimal ratio between the <i>PiggyBac</i> HER2-CAR-puro transposon and the Super <i>PiggyBac</i> transposase plasmid differed during the construction of the two HER2-targeted CAR-T cell types. The expansion abilities, CD3<sup>+</sup>CAR<sup>+</sup> population, CD4<sup>+</sup>CAR<sup>+</sup>/CD8<sup>+</sup>CAR<sup>+</sup> proportions, and memory and exhaustion markers between the two CAR-T groups were similar after using the optimized proportion of plasmid. Both CAR-T cell types exhibited significant antitumor activity, with the anti-HER2-13 CAR-T cells demonstrating superior target specificity. Therapeutic effects were observed with both CAR-T cells and trastuzumab in the MDA-MB-231<sup>HER2+</sup> breast tumor xenograft model, with anti-HER2-13 CAR-T cells demonstrating slightly enhanced efficacy and no evident off-target toxicity. <b>Conclusion:</b> These results highlight the potential of anti-HER2-13 CAR-T cells to serve as a safer and more efficacious alternative in HER2-targeted therapy.},
DOI = {10.32604/or.2025.065394}
}