In vitro study on reversible regulation of CAR-T function by Kinsenoside.

BACKGROUNDS: Chimeric antigen receptor T-cell (CAR-T) therapy has effectively treated various types of malignant tumors. However, effective methods to address its excessive expansion are currently lacking.

AIMS: We aimed to identify a natural compound that can safely and reversibly regulate CAR-T cell function.

MATERIALS AND

METHODS

This study combined virtual screening of network pharmacology with in vitro experimental verification to evaluate how a natural small-molecule kinsenoside (KD) can dynamically regulate the function of CAR-T cells. Human and CAR-T cells were treated with KD. Following the intervention and subsequent drug withdrawal, we conducted flow cytometry for T-cell functional tests, such as cell proliferation, cell cycle, and cytotoxicity. Additionally, transcriptome analysis was used to investigate the potential regulatory mechanisms involved.

RESULTS

The results indicated that KD inhibited CAR-T cell proliferation, blocked the cell cycle, reduced the killing function, and reduced IL6 secretion. KD drives CAR-T cells to Th17 fate without shifting CD4+/CD8 + ratios, modulating inflammation via Th17 pathways. The drug exhibits excellent characteristics, such as good reversibility, controllability, and low toxicity.

CONCLUSIONS

Thus, this natural occurring small molecule can act as a "functional switch," thereby providing new ideas for the balance between toxicity management and the efficacy of CAR-T therapy. Furthermore, it has the potential to pave the way for the combination of traditional Chinese medicine and modern cell therapy technology.