Adoptive T-cell therapy against solid tumours is limited by the apoptosis resistance mechanisms of tumour cells and by the extracellular, immunosuppressive tumour microenvironment. Here we report a temperature-sensitive genome-editing nanodevice that can deliver a Cas9 editor with an external trigger which can be used to edit the genome of tumour cells to reduce resistance to apoptosis and modulate the tumour microenvironment via a mild heating trigger. After local or systemic delivery of Cas9, mild heating is induced by non-invasive near-infrared (NIR) light or focused ultrasound (FUS) to activate Cas9, which initiates simultaneous genome editing of HSP70 (HSPA1A) and BAG3 in tumour cells. This disrupts the apoptotic resistance machinery of the tumour cells against adoptive T cells. At the same time, an NIR- or FUS-induced mild thermal effect reshapes the extracellular tumour microenvironment by disrupting the physical barriers and immune suppression. This facilitates the infiltration of adoptive T cells and enhances their therapeutic activity. Mild thermal Cas9 delivery is demonstrated in different murine tumour models which mimic a range of clinical indications, including a tumour model based on humanized patient-derived xenografts. As a result, the non-invasive thermal delivery of Cas9 significantly enhances the therapeutic efficacies of tumour-infiltrating lymphocytes and chimeric antigen receptor T and shows potential for clinical application.
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Source data are available for Figs. 1–6 and Supplementary Figures 2–28 in the associated source data files. The raw data of tumour transcriptome sequencing were deposited at the National Center for Biotechnology Information (NCBI) under BioProject: PRJNA916759. The raw data of deep sequencing were deposited at the NCBI under BioProject: PRJNA930575. Source data are provided with this paper.
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This work was supported by the National Key Research and Development Program of China (2021YFA0909900, Y.P.), the National Natural Science Foundation of China (82073779, 32261143727, Y.P.; U2001224, W.W.; U21A2097, K.L.) and the Natural Science Foundation of Zhejiang Province (Distinguished Young Scholar Program, LR21H300002, Y.P.). We thank H.-Y. Lai and T. He for their kind assistance with the FUS experiments.
Z.G. is a scientific cofounder of ZenCapsule and ZCapsule. Y.P. is a scientific cofounder of Ruidax. The other authors declare no competing interests.
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Chen, X., Wang, S., Chen, Y. et al. Non-invasive activation of intratumoural gene editing for improved adoptive T-cell therapy in solid tumours. Nat. Nanotechnol. (2023). https://doi.org/10.1038/s41565-023-01378-3