Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

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

Nature Nanotechnology volume 18, pages 933–944 (2023)Cite this article

Subjects

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.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

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.

Similar content being viewed by others

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.

References

  1. Hou, A. J., Chen, L. C. & Chen, Y. Y. Navigating CAR-T cells through the solid-tumor microenvironment. Nat. Rev. Drug Discov. 20, 531–550 (2021).

    Article  CAS  Google Scholar 

  2. Hong, M., Clubb, J. D. & Chen, Y. Y. Engineering CAR-T cells for next-generation cancer therapy. Cancer Cell 38, 473–488 (2020).

    Article  CAS  Google Scholar 

  3. Chen, J. et al. NR4A transcription factors limit CAR T cell function in solid tumors. Nature 567, 530–534 (2019).

    Article  CAS  Google Scholar 

  4. Schreiber, R. D., Old, L. J. & Smyth, M. J. Cancer immunoediting: integrating immunity’s roles in cancer suppression and promotion. Science 331, 1565–1570 (2011).

    Article  CAS  Google Scholar 

  5. Zou, W. Immunosuppressive networks in the tumor environment and their therapeutic relevance. Nat. Rev. Cancer 5, 263–274 (2005).

    Article  CAS  Google Scholar 

  6. Huang, Y. et al. Improving immune–vascular crosstalk for cancer immunotherapy. Nat. Rev. Immunol. 18, 195–203 (2018).

    Article  CAS  Google Scholar 

  7. Caruana, I. et al. Heparanase promotes tumor infiltration and antitumor activity of CAR-redirected T lymphocytes. Nat. Med. 21, 524–529 (2015).

    Article  CAS  Google Scholar 

  8. Chang, Z. L., Hou, A. J. & Chen, Y. Y. Engineering primary T cells with chimeric antigen receptors for rewired responses to soluble ligands. Nat. Protoc. 15, 1507–1524 (2020).

    Article  CAS  Google Scholar 

  9. Leen, A. M. et al. Reversal of tumor immune inhibition using a chimeric cytokine receptor. Mol. Ther. 22, 1211–1220 (2014).

    Article  CAS  Google Scholar 

  10. Cherkassky, L. et al. Human CAR T cells with cell-intrinsic PD-1 checkpoint blockade resist tumor-mediated inhibition. J. Clin. Invest. 126, 3130–3144 (2016).

    Article  Google Scholar 

  11. Liu, X. et al. A chimeric switch-receptor targeting PD1 augments the efficacy of second-generation CAR T-cells in advanced solid tumors. Cancer Res. 76, 1578–1590 (2016).

    Article  CAS  Google Scholar 

  12. Tang, T. C. Y., Xu, N. & Dolnikov, A. Targeting the immune-suppressive tumor microenvironment to potentiate CAR T cell therapy. Cancer Rep. Rev. 4, 1–5 (2020).

    Google Scholar 

  13. Karlsson, H. Approaches to augment CAR T-cell therapy by targeting the apoptotic machinery. Biochem. Soc. Trans. 44, 371–376 (2016).

    Article  CAS  Google Scholar 

  14. Green, D. R. The coming decade of cell death research: five riddles. Cell 177, 1094–1107 (2019).

    Article  CAS  Google Scholar 

  15. Jorgensen, I., Rayamajhi, M. & Miao, E. A. Programmed cell death as a defence against infection. Nat. Rev. Immunol. 17, 151–164 (2017).

    Article  CAS  Google Scholar 

  16. Kim, J. A., Kim, Y., Kwon, B. M. & Han, D. C. The natural compound cantharidin induces cancer cell death through inhibition of heat shock protein 70 (HSP70) and BCL2-associated athanogene domain 3 (BAG3) expression by blocking heat shock factor 1 (HSF1) binding to promoters. J. Biol. Chem. 288, 28713–28726 (2013).

    Article  CAS  Google Scholar 

  17. Rosati, A., Graziano, V., Laurenzi, V. D., Pascale, M. & Turco, M. C. BAG3: a multifaceted protein that regulates major cell pathways. Cell Death Dis. 2, e141 (2011).

    Article  CAS  Google Scholar 

  18. Wang, B. K. et al. Gold-nanorods–siRNA nanoplex for improved photothermal therapy by gene silencing. Biomaterials 78, 27 (2016).

    Article  CAS  Google Scholar 

  19. Joung, J. et al. CRISPR activation screen identifies BCL-2 proteins and B3GNT2 as drivers of cancer resistance to T cell-mediated cytotoxicity. Nat. Commun. 13, 1606 (2022).

    Article  CAS  Google Scholar 

  20. Rosati, A. et al. BAG3 promotes pancreatic ductal adenocarcinoma growth by activating stromal macrophages. Nat. Commun. 6, 8695 (2015).

    Article  CAS  Google Scholar 

  21. Lamprecht, A. Nanomedicines in gastroenterology and hepatology. Nat. Rev. Gastroenterol. Hepatol. 12, 669 (2015).

    Article  Google Scholar 

  22. Dudeja, V., Vickers, S. M. & Saluja, A. K. The role of heat shock proteins in gastrointestinal diseases. Gut 58, 1000–1009 (2009).

    Article  CAS  Google Scholar 

  23. Marzullo, L., Turco, M. C. & Marco, M. D. The multiple activities of BAG3 protein: mechanisms. Biochim. Biophys. Acta, Gen. Subj. 1864, 129628 (2020).

    Article  CAS  Google Scholar 

  24. Romano, M. F. et al. BAG3 protein controls B-chronic lymphocytic leukaemia cell apoptosis. Cell Death Differ. 10, 383–385 (2003).

    Article  CAS  Google Scholar 

  25. Ammirante, M. et al. IKKγ protein is a target of BAG3 regulatory activity in human tumor growth. Proc. Natl Acad. Sci. USA 107, 7497–7502 (2010).

    Article  CAS  Google Scholar 

  26. Eltoukhy, A. A., Chen, D., Albi, C. A., Langer, R. & Anderson, D. G. Degradable terpolymers with alkyl side chains demonstrate enhanced gene delivery potency and nanoparticle stability. Adv. Mater. 25, 1487–1493 (2013).

    Article  CAS  Google Scholar 

  27. Rui, Y. et al. High-throughput and high-content bioassay enables tuning of polyester nanoparticles for cellular uptake, endosomal escape, and systemic in vivo delivery of mRNA. Sci. Adv. 8, eabk2855 (2022).

    Article  CAS  Google Scholar 

  28. Zha, M. et al. An ester-substituted semiconducting polymer with efficient nonradiative decay enhances NIR-II photoacoustic performance for monitoring of tumor growth. Angew. Chem. Int. Ed. 59, 23268–23276 (2020).

    Article  CAS  Google Scholar 

  29. Banerjee, R., Tyagi, P., Li, S. & Huang, L. Anisamide-targeted stealth liposomes: a potent carrier for targeting doxorubicin to human prostate cancer cells. Int. J. Cancer 112, 693–700 (2004).

    Article  CAS  Google Scholar 

  30. Chen, Y. et al. Delivery of CRISPR/Cas9 plasmids by cationic gold nanorods: impact of the aspect ratio on genome editing and treatment of hepatic fibrosis. Chem. Mater. 33, 81–91 (2021).

    Article  CAS  Google Scholar 

  31. Li, N. et al. Chimeric antigen receptor-modified T cells redirected to EphA2 for the immunotherapy of non-small cell lung cancer. Transl. Oncol. 11, 11–17 (2018).

    Article  Google Scholar 

  32. Chen, X., Chen, Y., Xin, H., Wan, T. & Ping, Y. Near-infrared optogenetic engineering of photothermal nanoCRISPR for programmable genome editing. Proc. Natl Acad. Sci. USA 117, 2395–2405 (2020).

    Article  CAS  Google Scholar 

  33. Chen, Y., Yan, X. & Ping, Y. Optical manipulation of CRISPR/Cas9 functions: from ultraviolet to near-infrared light. ACS Mater. Lett. 2, 644–653 (2020).

    Article  CAS  Google Scholar 

  34. Zhang, W., He, M., Huang, G. & He, J. A comparison of ultrasound-guided high intensity focused ultrasound for the treatment of uterine fibroids in patients with an anteverted uterus and a retroverted uterus. Int. J. Hyperther. 32, 623–629 (2016).

    Article  Google Scholar 

  35. Klichinsky, M. et al. Human chimeric antigen receptor macrophages for cancer immunotherapy. Nat. Biotechnol. 38, 947–953 (2020).

    Article  CAS  Google Scholar 

  36. Guo, Y. et al. Metabolic reprogramming of terminally exhausted CD8+ T cells by IL-10 enhances anti-tumor immunity. Nat. Immunol. 22, 746–756 (2021).

    Article  CAS  Google Scholar 

  37. Etxeberria, I. et al. Intratumor adoptive transfer of IL-12 mRNA transiently engineered antitumor CD8+ T cells. Cancer Cell 36, 613–629 (2019).

    Article  CAS  Google Scholar 

  38. Singh, N. et al. Antigen-independent activation enhances the efficacy of 4-1BB-costimulated CD22 CAR T cells. Nat. Med. 27, 842–850 (2021).

    Article  CAS  Google Scholar 

  39. Etxeberria, I. et al. Engineering bionic T cells: signal 1, signal 2, signal 3, reprogramming and the removal of inhibitory mechanisms. Cell. Mol. Immunol. 17, 576–586 (2020).

    Article  CAS  Google Scholar 

  40. Rostamian, H. et al. A metabolic switch to memory CAR T cells: implications for cancer treatment. Cancer Lett. 500, 107–118 (2021).

    Article  CAS  Google Scholar 

  41. Korde, L. A., Somerfield, M. R. & Hershman, D. L. Use of immune checkpoint inhibitor pembrolizumab in the treatment of high-risk, early-stage triple-negative breast cancer: ASCO guideline rapid recommendation update. J. Clin. Oncol. 39, 1696–1698 (2021).

    Google Scholar 

  42. Yoshida, K., Yamaguchi, K., Okumura, N., Tanahashi, T. & Kodera, Y. Is conversion therapy possible in stage IV gastric cancer: the proposal of new biological categories of classification. Gastric Cancer 19, 329–338 (2016).

    Article  Google Scholar 

  43. Song, T., Lang, M., Ren, S., Gan, L. & Lu, W. The past, present and future of conversion therapy for liver cancer. Am. J. Cancer Res. 11, 4711–4724 (2021).

    CAS  Google Scholar 

  44. Sun, H. & Zhu, X. Downstaging conversion therapy in patients with initially unresectable advanced hepatocellular carcinoma: an overview. Front. Oncol. 11, 772195 (2021).

    Article  CAS  Google Scholar 

  45. Kishton, R. J., Lynn, R. C. & Restifo, N. P. Strength in numbers: identifying neoantigen targets for cancer immunotherapy. Cell 184, 5031–5052 (2021).

    Google Scholar 

  46. Storz, P. & Crawford, H. C. Carcinogenesis of pancreatic ductal adenocarcinoma. Gastroenterology 158, 2072–2081 (2020).

    Article  CAS  Google Scholar 

  47. Hosein, A. N., Dougan, S. K., Aguirre, A. J. & Maitra, A. Translational advances in pancreatic ductal adenocarcinoma therapy. Nat. Cancer 3, 272–286 (2022).

    Article  Google Scholar 

  48. Xue, G. et al. Adoptive cell therapy with tumor-specific Th9 cells induces viral mimicry to eliminate antigen-loss-variant tumor cells. Cancer Cell 39, 1610–1622 (2021).

    Article  CAS  Google Scholar 

  49. Hirabayashi, K. et al. Dual targeting CAR-T cells with optimal costimulation and metabolic fitness enhance antitumor activity and prevent escape in solid tumors. Nat. Cancer 2, 904–918 (2021).

    Article  CAS  Google Scholar 

  50. Bergers, G. & Fendt, S. The metabolism of cancer cells during metastasis. Nat. Rev. Cancer 21, 162–180 (2021).

    Article  CAS  Google Scholar 

Download references

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.

Author information

Author notes

  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

Authors
  1. Xiaohong Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Shuang Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yuxuan Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Huhu Xin
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Shuaishuai Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Di Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Yanan Xue
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Menglei Zha
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Hongjun Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Kai Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Zhen Gu
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Wei Wei
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Yuan Ping
    View author publications

    You can also search for this author in PubMed Google Scholar

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.

Ethics declarations

Competing interests

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.

Peer review

Peer review information

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

Additional information

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary information

Supplementary Information

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

Reporting Summary

Supplementary Data 1

Supplementary statistical source data.

Source data

Source Data Fig. 1

Unprocessed western blots and gels, and statistical data for Fig. 1.

Source Data Fig. 2

Unprocessed western blots and gels, and statistical data for Fig. 2.

Source Data Fig. 3

Statistical data for Fig. 3.

Source Data Fig. 4

Unprocessed western blots and gels, and statistical data for Fig. 4.

Source Data Fig. 5

Unprocessed western blots and gels, and statistical data for Fig. 5.

Source Data Fig. 6

Unprocessed western blots and gels, and statistical data for Fig. 6.

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

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. 18, 933–944 (2023). https://doi.org/10.1038/s41565-023-01378-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/s41565-023-01378-3

Search

Advanced search

Quick links

Nature Briefing: Translational Research

Sign up for the Nature Briefing: Translational Research newsletter — top stories in biotechnology, drug discovery and pharma.

Get what matters in translational research, free to your inbox weekly. Sign up for Nature Briefing: Translational Research