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Non-invasive activation of intratumoural gene editing for improved adoptive T-cell therapy in solid tumours

Nature Nanotechnology (2023)Cite this article

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Abstract

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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Fig. 1: Development of a LEGEND or FUGEND system that can potentiate the functions of adoptive T cells in solid tumours.
Fig. 2: LEGEND treatment reprograms the intracellular tumour resistance and the extracellular TME of solid tumours.
Fig. 3: LEGEND treatment potentiates intratumoural infiltration, proliferation and the antitumour effect of adoptive TILs.
Fig. 4: LEGEND treatment enhances the antitumour effect of adoptive CAR-T-cell therapy.
Fig. 5: LEGEND-sensitized adoptive EGFR-vIII-CAR-T-cell therapy in a humanized PDX model.
Fig. 6: FUGEND-sensitized adoptive TIL therapy by thermal-triggered genome editing in an orthotopic liver cancer mouse model.

Data availability

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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Acknowledgements

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.

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  1. These authors contributed equally: Xiaohong Chen, Shuang Wang, Yuxuan Chen.

Authors and Affiliations

  1. College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, China

    Xiaohong Chen, Yuxuan Chen, Huhu Xin, Shuaishuai Zhang, Di Wu, Yanan Xue, Hongjun Li, Zhen Gu & Yuan Ping

  2. State Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing, China

    Shuang Wang & Wei Wei

  3. School of Chemical Engineering, University of Chinese Academy of Sciences, Beijing, China

    Shuang Wang & Wei Wei

  4. Department of General Surgery, Sir Run Run Shaw Hospital, School of Medicine, Zhejiang University, Hangzhou, China

    Yanan Xue & Zhen Gu

  5. Department of Biomedical Engineering, Southern University of Science and Technology, Shenzhen, China

    Menglei Zha & Kai Li

  6. Liangzhu Laboratory, Zhejiang University Medical Center, Hangzhou, China

    Hongjun Li, Zhen Gu & Yuan Ping

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Contributions

X.C., S.W. and Y.C. designed and performed experiments and analysed and interpreted data. H.X. and S.Z. constructed the plasmid and analysed deep-sequencing results. Y.X. helped construct the PDX model. M.Z. and K.L. synthesized the BDT-TQE semiconducting polymer. D.W., H.L. and Z.G. discussed the manuscript. W.W. and Y.P. conceived the idea, designed the study, analysed and interpreted data, and wrote the manuscript.

Corresponding authors

Correspondence to Wei Wei or Yuan Ping.

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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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Nature Nanotechnology thanks Kanyi Pu and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

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Supplementary methods, figs. 1–28, tables 1–3, references, and unprocessed blots and gels for supplementary figures.

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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

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