Intratumoral administration of cGAMP transiently accumulates potent macrophages for anti-tumor immunity at a mouse tumor site (original) (raw)

Abstract

Stimulator of IFN genes (STING) spontaneously contributes to anti-tumor immunity by inducing type I interferons (IFNs) following sensing of tumor-derived genomic DNAs in the tumor-bearing host. Although direct injection of STING ligands such as cyclic diguanylate monophosphate (c-di-GMP) and cyclic [G(2′,5′)pA(3′,5′)p] (cGAMP) into the tumor microenvironment exerts anti-tumor effects through strong induction of type I IFNs and activation of innate and adaptive immunity, the precise events caused by STING in the tumor microenvironment remain to be elucidated. We describe here our finding that a CD45+ CD11bmid Ly6C+ cell subset transiently accumulated in mouse tumor microenvironment of 4T1 breast cancer, squamous cell carcinomas, CT26 colon cancer, or B16F10 melanoma tissue after intratumoral injection of cGAMP. The accumulated cells displayed a macrophage (M ) phenotype since the cells were positive for F4/80 and MHC class II and negative for Ly6G. Intratumoral cGAMP treatment did not induce Mφ accumulation in STING-deficient mice. Depletion of CD8+ T cell using anti-CD8 mAb impaired the anti-tumor effects of cGAMP treatment. Depletion of the Mφ using clodronate liposomes impaired the anti-tumor effects of cGAMP treatment. Functional analysis indicated that the STING-triggered tumor-migrating Mφ exhibited phagocytic activity, production of tumor necrosis factor alpha TNFα), and high expression levels of T cell-recruiting chemokines, Cxcl10 and Cxcl11, IFN-induced molecules, MX dynamin-like GTPase 1 (Mx1) and 2′-5′ oligoadenylate synthetase-like 1 (Oasl1), nitric oxide synthase 2 (Nos2), and interferon beta 1 (Ifnb1). These results indicate that the STING-triggered tumor-migrating Mφ participate in the anti-tumor effects of STING-activating compounds.

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Abbreviations

c-di-GMP:

Cyclic diguanylate monophosphate

cGAMP:

Cyclic [G(2′,5′)pA(3′,5′)p]

DCs:

Dendritic cells

IFNb1:

Interferon beta 1

IFNγ:

Interferon gamma

IFNs:

Interferons

IL:

Interleukin

LPS:

Lipopolysaccharide

MDSCs:

Myeloid-derived suppressor cells

Mφ:

Macrophage(s)

MO-MDSCs:

Monocytic MDSCs

Mx1:

MX dynamin-like GTPase 1

Nos2:

Nitric oxide synthase 2

Oasl1:

2′-5′ Oligoadenylate synthetase-like 1

PD-1:

Programmed death-1

STING:

Stimulator of interferon genes

TAM:

Tumor-associated Mφ

TILs:

Tumor-infiltrating leukocytes

TNFα:

Tumor necrosis factor alpha

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Acknowledgements

The authors thank Mr. Hayakawa Toshiyuki and Ms. Hino Chihiro (at the Animal Laboratory for Medical Research, Center for Advanced Research and Education, Asahikawa Medical University) and Ms. Matsumoto Rie (at the Department of Pathology, Asahikawa Medical University) for devotedly maintaining the mice and Mr. Akutsu Hiroaki (at Center for Advanced Research and Education, Asahikawa Medical University) for technically supporting cell sorting.

This work was supported by grants from Japan Society for the Promotion of Science KAKENHI Grant Number 16K19070 (Ohkuri) and 16K19071 (Kosaka), the Akiyama Life Science Foundation (Ohkuri), and Innovative Research in Life Science from Asahikawa Medical University (Ohkuri and Kosaka).

Author contributions

Ohkuri and Kosaka preformed experiments, analyzed results, and made the figures; Ohkuri, Kosaka, Ishibashi, Kumai, Hirata, Ohara, Nagato, Oikawa, Aoki, Harabuchi, Celis, and Kobayashi discussed the results; Ohkuri, Kosaka, Celis, and Kobayashi designed the research and wrote the paper.

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Author notes

  1. T. Ohkuri and A. Kosaka contributed equally to this article.

Authors and Affiliations

  1. Department of Pathology, Asahikawa Medical University, Midorigaoka-Higashi 2-1-1, Asahikawa, 078-8510, Japan
    Takayuki Ohkuri, Akemi Kosaka, Kei Ishibashi, Takumi Kumai, Yui Hirata, Kenzo Ohara, Kensuke Oikawa, Naoko Aoki & Hiroya Kobayashi
  2. Respiratory and Breast Center, Asahikawa Medical University Hospital, Asahikawa, 078-8510, Japan
    Kei Ishibashi
  3. Department of Otolaryngology, Head and Neck Surgery, Asahikawa Medical University, Asahikawa, 078-8510, Japan
    Takumi Kumai, Yui Hirata, Kenzo Ohara, Toshihiro Nagato & Yasuaki Harabuchi
  4. Cancer Immunology, Inflammation and Tolerance Program, Augusta University GRU Cancer Center, Augusta, GA, 30912, USA
    Takumi Kumai & Esteban Celis

Authors

  1. Takayuki Ohkuri
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  2. Akemi Kosaka
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  3. Kei Ishibashi
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  4. Takumi Kumai
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  5. Yui Hirata
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  6. Kenzo Ohara
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  7. Toshihiro Nagato
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  8. Kensuke Oikawa
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  9. Naoko Aoki
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  10. Yasuaki Harabuchi
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  11. Esteban Celis
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  12. Hiroya Kobayashi
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Correspondence toTakayuki Ohkuri or Hiroya Kobayashi.

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Ohkuri, T., Kosaka, A., Ishibashi, K. et al. Intratumoral administration of cGAMP transiently accumulates potent macrophages for anti-tumor immunity at a mouse tumor site.Cancer Immunol Immunother 66, 705–716 (2017). https://doi.org/10.1007/s00262-017-1975-1

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