Pluripotent stem cell-derived siTNK cells attack tumors via a synthetic CD8-CD28 TCR complex

Zheng, X.; Zhang, F.; Lin, Y.; Hu, F.; Weng, Q.; Liu, P.; Wang, Z.; Zhang, C.; Liu, Y.; Liu, L.; Zhu, Y.; Xiao, Z.; Wang, Y.; Zhang, L.; Qi, H.; Shen, Y.; Chen, Y.; Wu, J.; Xu, J.; Zhao, Y.; Wang, T.; Huang, D.; Xia, C.; Wang, J.; Zhang, M.

doi:10.1101/2025.07.11.663392

bioRxiv · 10.1101/2025.07.11.663392

Pluripotent stem cell-derived siTNK cells attack tumors via a synthetic CD8-CD28 TCR complex

Zheng, X.·Zhang, F.·Lin, Y.·Hu, F.·Weng, Q.·Liu, P.·Wang, Z.·Zhang, C.·Liu, Y.·Liu, L.·Zhu, Y.·Xiao, Z.·Wang, Y.·Zhang, L.·Qi, H.·Shen, Y.·Chen, Y.·Wu, J.·Xu, J.·Zhao, Y.·Wang, T.·Huang, D.·Xia, C.·Wang, J.·Zhang, M.

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Abstract

Tumor-associated antigen-specific T cell receptor (TCR)-engineered T cells offer a promising strategy for cancer therapy. Natural killer (NK) cells exhibit broad anti-tumor activity with low side effects but lack the capacity to recognize intracellular antigens. Here, we found that the human pluripotent stem cell (hPSC)-derived iNK cells, unlike tissue-isolated NK cells, expressed all four CD3 subunits at the transcriptome level. We introduced a synthetic gene-expressing complex (SCOTR), encoding a tumor antigen- specific TCR, CD8 coreceptor, and CD28 costimulatory molecule, into hPSCs to generate SCOTR-hPSCs. The SCOTR-hPSCs gave rise to abundant synthetic TCR complex-expressing iNK (siTNK) cells via an organoid induction method. These siTNK cells showed HLA-dependent, antigen-specific cytotoxicity against tumor cells and significantly suppressed tumor growth in tumor xenograft animal models, while also preserving universal non-specific tumor-killing activity. Collectively, siTNK cells, as hPSC-derived hybrid cells with dual features of adaptive T and inherent NK cells, offer an artificial cell source for human immunotherapies.

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Zheng, X., Zhang, F., Lin, Y., Hu, F., Weng, Q., Liu, P., Wang, Z., Zhang, C., Liu, Y., Liu, L., Zhu, Y., Xiao, Z., Wang, Y., Zhang, L., Qi, H., Shen, Y., Chen, Y., Wu, J., Xu, J., Zhao, Y., Wang, T., Huang, D., Xia, C., Wang, J., Zhang, M.. 2025-07-12. Pluripotent stem cell-derived siTNK cells attack tumors via a synthetic CD8-CD28 TCR complex. https://doi.org/10.1101/2025.07.11.663392

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