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Combination of CHEK1/2 inhibition and ionizing radiation results in abscopal tumor response through increased micronuclei formation

Oncogene volume 39, pages 4344–4357 (2020)Cite this article

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Abstract

We explore a novel strategy of activating immune signaling through increased micronuclei formation utilizing a cell cycle checkpoint inhibitor to drive cell cycle progression following ionizing radiation. The Chk1/2 inhibitor AZD7762 is used to abrogate radiation therapy (RT)-induced G2/M cell cycle arrest in multiple cell lines and, we find that this therapeutic combination promotes increased micronuclei formation in vitro and subsequently drives increased type I interferon signaling and cytotoxic T-cell activation. In vivo studies using B16-F10 melanoma cancer cells implanted in C57/BL6 mice demonstrate improved rates of tumor control at the abscopal (unirradiated) site, located outside of the radiation field, only in the AZD7762 + RT group, with a corresponding reduction in mean tumor volume, increase in the CD8 T-cell population, and immune activated gene signaling. Our results demonstrate that targeted inhibition of cell cycle checkpoint activation following ionizing radiation drives increased production of immunogenic micronuclei, leading to systemic tumor response with potential future clinical benefit.

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Fig. 1: AZD7762 treatment abrogates RT induced G2/M Checkpoint.
Fig. 2: AZD7762 + RT combination treatment increases micronuclei formation.
Fig. 3: cGAS co-localizes with induced micronuclei.
Fig. 4: AZD7762 + RT combination increases Type I interferon regulated gene and protein expression and is mediated through the STING pathway.
Fig. 5: Secreted IFN-β protein expression by ELISA in B16F10 tumor cells.
Fig. 6: Tumor response in implanted B16-F10 tumors.
Fig. 7: Tumor volume in implanted B16-F10 tumors.
Fig. 8: Immune activation by CD8 T-cell recruitment and gene expression in implanted B16-F10 tumors.

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Acknowledgements

This study was supported by NIH/NCI grant (1R01CA182747 (PI: AM, CK) and the University of Pennsylvania Radiation Oncology Resident Research Award (to H-HC). The authors thank Roger Greenberg group for contribution of cGAS constructs.

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  1. These authors contributed equally: Hann-Hsiang Chao, Ilias V. Karagounis

Authors and Affiliations

  1. Department of Radiation Oncology, University of Pennsylvania, Philadelphia, PA, USA

    Hann-Hsiang Chao, Ilias V. Karagounis, Christoforos Thomas, Noëlle B. François, Andrea Facciabene, Constantinos Koumenis & Amit Maity

  2. Department of Radiation Oncology, McGuire VA Medical Center, Richmond, VA, USA

    Hann-Hsiang Chao

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Contributions

Conception and design: H-HC, I-VK, CT and AM. Development of methodology: H-HC, I-VK, CT, AF, CK and AM. Acquisition of data (provided animals, acquired, and managed patients, provided facilities, etc.): H-HC, I-VK, CT, and NBF. Analysis and interpretation of data (e.g., statistical analysis, biostatistics, and computational analysis): H-HC, I-VK. Writing, review, and/or revision of the manuscript: H-HC, I-VK, CT, NBF, AF, CK and AM. Administrative, technical, or material support (i.e., reporting or organizing data, constructing databases): H-HC, I-VK and NBF. Study supervision: AM.

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Correspondence to Amit Maity.

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AM reports receiving research funding from Merck. No potential competing interests are reported by the other authors.

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Chao, HH., Karagounis, I.V., Thomas, C. et al. Combination of CHEK1/2 inhibition and ionizing radiation results in abscopal tumor response through increased micronuclei formation. Oncogene 39, 4344–4357 (2020). https://doi.org/10.1038/s41388-020-1300-x

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