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:

Tumor cells suppress radiation-induced immunity by hijacking caspase 9 signaling

Nature Immunology volume 21, pages 546–554 (2020)Cite this article

Subjects

An Author Correction to this article was published on 16 September 2020

This article has been updated

Abstract

High-dose radiation activates caspases in tumor cells to produce abundant DNA fragments for DNA sensing in antigen-presenting cells, but the intrinsic DNA sensing in tumor cells after radiation is rather limited. Here we demonstrate that irradiated tumor cells hijack caspase 9 signaling to suppress intrinsic DNA sensing. Instead of apoptotic genomic DNA, tumor-derived mitochondrial DNA triggers intrinsic DNA sensing. Specifically, loss of mitochondrial DNA sensing in Casp9−/− tumors abolishes the enhanced therapeutic effect of radiation. We demonstrated that combining emricasan, a pan-caspase inhibitor, with radiation generates synergistic therapeutic effects. Moreover, loss of CASP9 signaling in tumor cells led to adaptive resistance by upregulating programmed death-ligand 1 (PD-L1) and resulted in tumor relapse. Additional anti-PD-L1 blockade can further overcome this acquired immune resistance. Therefore, combining radiation with a caspase inhibitor and anti-PD-L1 can effectively control tumors by sequentially blocking both intrinsic and extrinsic inhibitory signaling.

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: Tumor cells hijack intrinsic apoptosis to restrict type I IFN production after radiation.
Fig. 2: Tumor-intrinsic CASP9 signaling suppresses radiation-induced antitumor immunity.
Fig. 3: Blocking CASP9 signaling to facilitate tumor-intrinsic mtDNA sensing after radiation.
Fig. 4: Casp9−/− tumor-derived dsDNA innate sensing is required for provoking radiation-mediated antitumor immunity.
Fig. 5: Tumors evolve resistance to adaptive immunity by upregulating PD-L1.
Fig. 6: Emricasan synergizes with radiation and anti-PD-L1.

Similar content being viewed by others

Data availability

All data supporting the findings of this study are available within the article and its supplementary information files and from the corresponding author upon reasonable request. A reporting summary for this article is available as a Supplementary Information file.

Change history

  • 16 September 2020

    An amendment to this paper has been published and can be accessed via a link at the top of the paper.

References

  1. Deng, L. et al. STING-dependent cytosolic DNA sensing promotes radiation-induced type I interferon-dependent antitumor immunity in immunogenic tumors. Immunity 41, 843–852 (2014).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Burnette, B. C. et al. The efficacy of radiotherapy relies upon induction of type I interferon-dependent innate and adaptive immunity. Cancer Res. 71, 2488–2496 (2011).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Vanpouille-Box, C. et al. DNA exonuclease Trex1 regulates radiotherapy-induced tumour immunogenicity. Nat. Commun. 8, 15618 (2017).

    Article  PubMed  PubMed Central  Google Scholar 

  4. Harding, S. M. et al. Mitotic progression following DNA damage enables pattern recognition within micronuclei. Nature 548, 466 (2017).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Li, P. et al. Cytochrome c and dATP-dependent formation of Apaf-1/caspase-9 complex initiates an apoptotic protease cascade. Cell 91, 479–489 (1997).

    Article  CAS  PubMed  Google Scholar 

  6. Maier, P., Hartmann, L., Wenz, F. & Herskind, C. Cellular pathways in response to ionizing radiation and their targetability for tumor radiosensitization. Int. J. Mol. Sci. 17, 102 (2016).

    Article  PubMed Central  CAS  Google Scholar 

  7. Gudkov, A. V. & Komarova, E. A. The role of p53 in determining sensitivity to radiotherapy. Nat. Rev. Cancer 3, 117 (2003).

    Article  CAS  PubMed  Google Scholar 

  8. Rongvaux, A. et al. Apoptotic caspases prevent the induction of type I interferons by mitochondrial DNA. Cell 159, 1563–1577 (2014).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. White, M. J. et al. Apoptotic caspases suppress mtDNA-induced STING-mediated type I IFN production. Cell 159, 1549–1562 (2014).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. McArthur, K. et al. BAK/BAX macropores facilitate mitochondrial herniation and mtDNA efflux during apoptosis. Science 359, eaao6047 (2018).

    Article  PubMed  CAS  Google Scholar 

  11. Lama, L. et al. Development of human cGAS-specific small-molecule inhibitors for repression of dsDNA-triggered interferon expression. Nat. Commun. 10, 2261 (2019).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  12. Hauff, P., Gottwald, U. & Ocker, M. Early to phase II drugs currently under investigation for the treatment of liver fibrosis. Expert Opin. Investig. Drugs 24, 309–327 (2015).

    Article  CAS  PubMed  Google Scholar 

  13. Mehta, G. et al. A placebo-controlled, multicenter, double-blind, phase 2 randomized trial of the pan-caspase inhibitor Emricasan in patients with acutely decompensated cirrhosis. J. Clin. Exp. Hepatol. 8, 224–234 (2018).

    Article  PubMed  Google Scholar 

  14. Galluzzi, L., Buque, A., Kepp, O., Zitvogel, L. & Kroemer, G. Immunological effects of conventional chemotherapy and targeted anticancer agents. Cancer Cell 28, 690–714 (2015).

    Article  CAS  PubMed  Google Scholar 

  15. Liu, X. et al. CD47 blockade triggers T cell-mediated destruction of immunogenic tumors. Nat. Med. 21, 1209 (2015).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Stagg, J. et al. Anti-ErbB-2 mAb therapy requires type I and II interferons and synergizes with anti-PD-1 or anti-CD137 mAb therapy. Proc. Natl Acad. Sci. USA 108, 7142–7147 (2011).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Liu, Z. et al. Hypofractionated EGFR tyrosine kinase inhibitor limits tumor relapse through triggering innate and adaptive immunity. Sci. Immunol. 4, eaav6473 (2019).

    Article  CAS  PubMed  Google Scholar 

  18. Lee, Y. et al. Therapeutic effects of ablative radiation on local tumor require CD8+ T cells: changing strategies for cancer treatment. Blood 114, 589–595 (2009).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Lim, J. Y., Gerber, S. A., Murphy, S. P. & Lord, E. M. Type I interferons induced by radiation therapy mediate recruitment and effector function of CD8+ T cells. Cancer Immunol. Immunother. 63, 259–271 (2014).

    Article  CAS  PubMed  Google Scholar 

  20. Roberts, E. W. et al. Critical role for CD103+/CD141+ dendritic cells bearing CCR7 for tumor antigen trafficking and priming of T cell immunity in melanoma. Cancer Cell 30, 324–336 (2016).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Spranger, S., Dai, D., Horton, B. & Gajewski, T. F. Tumor-residing Batf3 dendritic cells are required for effector T cell trafficking and adoptive T cell therapy. Cancer Cell 31, 711–723. e714 (2017).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Hildner, K. et al. Batf3 deficiency reveals a critical role for CD8α+ dendritic cells in cytotoxic T cell immunity. Science 322, 1097–1100 (2008).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Dou, Z. et al. Cytoplasmic chromatin triggers inflammation in senescence and cancer. Nature 550, 402 (2017).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Mackenzie, K. J. et al. cGAS surveillance of micronuclei links genome instability to innate immunity. Nature 548, 461 (2017).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Ferguson, B. J., Mansur, D. S., Peters, N. E., Ren, H. & Smith, G. L. DNA-PK is a DNA sensor for IRF-3-dependent innate immunity. eLife 1, e00047 (2012).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  26. Liu, H. et al. Nuclear cGAS suppresses DNA repair and promotes tumorigenesis. Nature 563, 131 (2018).

    Article  CAS  PubMed  Google Scholar 

  27. Yang, H., Wang, H., Ren, J., Chen, Q. & Chen, Z. J. cGAS is essential for cellular senescence. Proc. Natl Acad. Sci. USA 114, E4612–E4620 (2017).

    CAS  PubMed  PubMed Central  Google Scholar 

  28. Rodríguez-Ruiz, M. E., Vanpouille-Box, C., Melero, I., Formenti, S. C. & Demaria, S. Immunological mechanisms responsible for radiation-induced abscopal effect. Trends Immunol. 39, 644–655 (2018).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  29. Dovedi, S. & Illidge, T. The antitumor immune response generated by fractionated radiation therapy may be limited by tumor cell adaptive resistance and can be circumvented by PD-L1 blockade. Oncoimmunology 4, e1016709 (2015).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Benci, J. L. et al. Tumor interferon signaling regulates a multigenic resistance program to immune checkpoint blockade. Cell 167, 1540–1554 (2016).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Deng, L., Liang, H., Burnette, B., weicheslbaum, r & Fu, Y.-X. Radiation and anti-PD-L1 antibody combinatorial therapy induces T cell-mediated depletion of myeloid-derived suppressor cells and tumor regression. Oncoimmunology 3, e28499 (2014).

    Article  PubMed  PubMed Central  Google Scholar 

  32. Zou, W., Wolchok, J. D. & Chen, L. PD-L1 (B7-H1) and PD-1 pathway blockade for cancer therapy: mechanisms, response biomarkers, and combinations. Sci. Transl. Med. 8, 328rv324–328rv324 (2016).

    Article  CAS  Google Scholar 

  33. Cai, X., Chiu, Y.-H. & Chen, Z. J. The cGAS-cGAMP-STING pathway of cytosolic DNA sensing and signaling. Mol. Cell 54, 289–296 (2014).

    Article  CAS  PubMed  Google Scholar 

  34. Wang, H. et al. cGAS is essential for the antitumor effect of immune checkpoint blockade. Proc. Nat. Acad. Sci. USA 114, 1637–1642 (2017).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Xu, M. M. et al. Dendritic cells but not macrophages sense tumor mitochondrial DNA for cross-priming through signal regulatory protein α signaling. Immunity 47, 363–373 (2017).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Glück, S. et al. Innate immune sensing of cytosolic chromatin fragments through cGAS promotes senescence. Nat. Cell Biol. 19, 1061 (2017).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  37. Andreeva, L. et al. cGAS senses long and HMGB/TFAM-bound U-turn DNA by forming protein–DNA ladders. Nature 549, 394 (2017).

    Article  CAS  PubMed  Google Scholar 

  38. Kim, K. W., Moretti, L. & Lu, B. M867, a novel selective inhibitor of caspase-3 enhances cell death and extends tumor growth delay in irradiated lung cancer models. PLoS ONE 3, e2275 (2008).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  39. Werthmöller, N., Frey, B., Wunderlich, R., Fietkau, R. & Gaipl, U. Modulation of radiochemoimmunotherapy-induced B16 melanoma cell death by the pan-caspase inhibitor zVAD-fmk induces anti-tumor immunity in a HMGB1-, nucleotide- and T cell-dependent manner. Cell Death Dis. 6, e1761 (2015).

    Article  PubMed  PubMed Central  Google Scholar 

  40. Brumatti, G. et al. The caspase-8 inhibitor Emricasan combines with the SMAC mimetic birinapant to induce necroptosis and treat acute myeloid leukemia. Sci. Transl. Med. 8, 339ra369 (2016).

    Article  CAS  Google Scholar 

  41. Giampazolias, E. et al. Mitochondrial permeabilization engages NF-κB-dependent anti-tumour activity under caspase deficiency. Nat. Cell Biol. 19, 1116 (2017).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. Rodriguez-Ruiz, M. E. et al. Apoptotic caspases inhibit abscopal responses to radiation and identify a new prognostic biomarker for breast cancer patients. Oncoimmunology 8, e1655964 (2019).

    Article  PubMed  PubMed Central  Google Scholar 

  43. Bai, M. et al. In vivo cell kinetics in breast carcinogenesis. Breast Cancer Res. 3, 276 (2001).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  44. Huang, J.-S. et al. Caspase-3 expression in tumorigenesis and prognosis of buccal mucosa squamous cell carcinoma. Oncotarget 8, 84237 (2017).

    Article  PubMed  PubMed Central  Google Scholar 

  45. Flanagan, L. et al. Low levels of caspase-3 predict favourable response to 5FU-based chemotherapy in advanced colorectal cancer: caspase-3 inhibition as a therapeutic approach. Cell Death Dis. 7, e2087 (2017).

    Article  CAS  Google Scholar 

  46. Zhang, Z. et al. Increased HMGB1 and cleaved caspase-3 stimulate the proliferation of tumor cells and are correlated with the poor prognosis in colorectal cancer. J. Exp. Clin. Cancer Res. 34, 51 (2015).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  47. Hu, Q. et al. Elevated cleaved caspase-3 is associated with shortened overall survival in several cancer types. Int. J. Clin. Exp. Pathol. 7, 5057 (2014).

    PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

We thank R. W. Welchselbaum for providing reagents and assisting with experiments and the UT southwestern Flow Cytometry Facility and Animal Resources Center. YXF holds the Mary Nell and Ralph B. Rogers Professorship in Immunology. This work was supported by NCI CA134563, Texas CPRIT grant RR150072 and RR180725 (established CPRIT scholar in cancer research) to Y.-X. F.

Author information

Author notes

  1. These authors contributed equally: Chuanhui Han, Zhida Liu.

Authors and Affiliations

  1. Department of Pathology, UT Southwestern Medical Center, Dallas, TX, USA

    Chuanhui Han, Zhida Liu, Aijun Shen, Chunbo Dong, Anli Zhang, Casey Moore, Zhenhua Ren, Changzheng Lu, Xuezhi Cao, Jian Qiao & Yang-Xin Fu

  2. Department of Molecular Biology, UT Southwestern Medical Center, Dallas, TX, USA

    Yunjia Zhang & Chun-Li Zhang

  3. Department of Immunology, UT Southwestern Medical Center, Dallas, TX, USA

    Casey Moore & Yang-Xin Fu

Authors
  1. Chuanhui Han
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Zhida Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yunjia Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Aijun Shen
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Chunbo Dong
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Anli Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Casey Moore
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Zhenhua Ren
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Changzheng Lu
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Xuezhi Cao
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Chun-Li Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Jian Qiao
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Yang-Xin Fu
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

C.H., Z.L. and Y.-X.F. designed experiments and analyzed data. C.H. and Z.L. performed experiments. C.H. and Y.-X.F. wrote the manuscript. Z.L. J.Q. and C.M. revised the manuscript. Y.Z. and C.-L.Z. helped with SIM imaging. A.S., C.D., A.Z., Z.R., C.L. and X.C. provided mice and reagents. J.Q. gave valuable advice. Y.-X.F. supervised the project.

Corresponding authors

Correspondence to Jian Qiao or Yang-Xin Fu.

Ethics declarations

Competing interests

The authors declare no competing interests.

Additional information

Editor recognition statement: Zoltan Fehervari was the primary editor on this article and managed its editorial process and peer review in collaboration with the rest of the editorial team.

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

Supplementary information

Supplementary Information

Supplementary Figs. 1–6 and Supplementary Table 1.

Reporting Summary

Source data

Source Data Fig. 1

Statistical source data for Fig. 1.

Source Data Fig. 2

Statistical source data for Fig. 2.

Source Data Fig. 3

Statistical source data for Fig. 3.

Source Data Fig. 4

Statistical source data for Fig. 4.

Source Data Fig. 5

Statistical source data for Fig. 5.

Source Data Fig. 6

Statistical source data for Fig. 6.

Source Data Supplementary Data Fig. 1

Statistical source data for supplementary Fig. 1.

Source Data Supplementary Data Fig. 2

Statistical source data for supplementary Fig. 2.

Source Data Supplementary Data Fig. 3

Statistical source data for supplementary Fig. 3.

Source Data Supplementary Data Fig. 4

Statistical source data for supplementary Fig. 4.

Source Data Supplementary Data Fig. 5

Statistical source data for supplementary Fig. 5.

Source Data Supplementary Data Fig. 6

Statistical source data for supplementary Fig. 6.

Source Data Fig. 1e

Unprocessed western blot images for Fig. 1e.

Source Data Supplementary Data Fig. 4a

Unprocessed western blot images for Supplementary Fig. 4a.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Han, C., Liu, Z., Zhang, Y. et al. Tumor cells suppress radiation-induced immunity by hijacking caspase 9 signaling. Nat Immunol 21, 546–554 (2020). https://doi.org/10.1038/s41590-020-0641-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/s41590-020-0641-5

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing