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Activation of TLR4 signaling inhibits progression of osteosarcoma by stimulating CD8-positive cytotoxic lymphocytes

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Abstract

Background

Osteosarcoma (OS) is the most common malignant bone tumor and the prognosis of advanced cases is still poor. Recently, there have been several reports suggesting the relationship between innate immunity and OS, but the detailed mechanism is unknown. We demonstrate the relationship between OS and Toll-like receptor 4 (TLR4) which is one of the most important factors in innate immunity.

Methods

We established a syngenic mouse tumor model using C3H/HeN, C3H/HeJ mouse and a highly metastatic OS cell line, LM8. TLR4 activation with lipopolysaccharide (LPS) was performed on both mice and its influence on the progression of OS was evaluated. We also performed CD8 + cells depletion to examine the influence on TLR4 activation effects.

Results

Tumor volume of C3H/HeN mice was significantly smaller and overall survival of C3H/HeN mice was significantly longer than C3H/HeJ mice. We found more CD8+ cells infiltrating in lung metastases of C3H/HeN mice and depletion of CD8+ cells canceled the antitumor effects of LPS.

Conclusion

TLR4 activation by LPS increased CD8+ cells infiltrating into lung metastases and suppressed OS progression in the mouse model. TLR4 activation may suppress the progression of OS via stimulating CD8+ cells and can be expected as a novel treatment for OS.

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Abbreviations

CLP:

Liposomal clodronate

LPS:

Lipopolysaccharide

OAS:

Overall survival

OS:

Osteosarcoma

PFS:

Progression-free survival

S.D:

Standard deviation

TAM:

Tumor-associated macrophage

TLR:

Toll-like receptor

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Acknowledgements

We thank Dr. Tetsuzo Tagawa from Department of Surgery and Science, Graduate School of Medical Sciences, Kyushu University for kindly providing clinical samples of the patients with OS.

Funding

This work was supported by Grants-in-Aid for Young Scientists (18K16627, 19K16802), Grant-in-Aid for Scientific Research (18K09067) from the Japan Society for the Promotion of Science, a Grant from Japan Orthopedics and Traumatology Research Foundation Inc. (No. 332). Toshifumi Fujiwara was also recipient of Fukuoka Foundation for Sound Health Cancer Research Fund.

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Authors and Affiliations

  1. Department of Orthopaedic Surgery, Graduate School of Medical Sciences, Kyushu University, 3-1-1, Maidashi, Higashi-ku, Fukuoka, Fukuoka, Japan

    Kenichiro Yahiro, Yoshihiro Matsumoto, Hisakata Yamada, Makoto Endo, Nokitaka Setsu, Toshifumi Fujiwara, Makoto Nakagawa, Atsushi Kimura, Eijiro Shimada & Yasuharu Nakashima

  2. Division of Orthopaedic Surgery, National Cancer Center Hospital, 5-1-1, Tsukiji, Chuo-ku, Tokyo, Japan

    Makoto Nakagawa

  3. Department of Immunobiology and Neuroscience Medical. Medical Institute of Bioregulation, Kyushu University, 3-1-1, Maidashi, Higashi-ku, Fukuoka, Fukuoka, Japan

    Seiji Okada

  4. Department of Anatomic Pathology, Pathological Sciences, Graduate School of Medical Sciences, Kyushu University, 3-1-1, Maidashi, Higashi-ku, Fukuoka, Fukuoka, Japan

    Yoshinao Oda

Authors
  1. Kenichiro Yahiro
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  2. Yoshihiro Matsumoto
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  3. Hisakata Yamada
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  6. Toshifumi Fujiwara
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  7. Makoto Nakagawa
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  8. Atsushi Kimura
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  9. Eijiro Shimada
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  11. Yoshinao Oda
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  12. Yasuharu Nakashima
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Contributions

KY, YM, and HY designed research; KY, AK, and ES performed research; YM, HY, EM, NS, and TF analyzed data; MN, SO, YO, and YN supervised research; KY and YM drafted the manuscript.

Corresponding author

Correspondence to Yoshihiro Matsumoto.

Ethics declarations

Conflict of interest

The authors declare no conflict of interest.

Ethics approval and ethical standards

The study was conducted in accordance with the Declaration of Helsinki and approved by the Institutional Ethical Review Board of the Kyushu University (#27–420, 2016/3/31). The animal study was approved by the Institutional Ethical Review Board of the Kyushu University (#A28-160–0, 2017–3-31). Experiments involving animals were performed in compliance with the guidelines established by the Animal Care and Use Committee and this study was approved by the Institutional Review Board at Kyushu University.

Informed consent

Prior to the use of patient data and specimens in the study, written informed consent was obtained from all patients. They all agreed to use their clinical data and bio materials for this research and publication. Written informed consent was obtained also from the parents when the patient was a minor.

Animal source

C3H/HeN and C3H/HeJ mice were purchased from Kyudo (Saga, Japan).

Cell line authentication

LM8 mouse osteosarcoma cells were purchased from RIKEN BioResource Research Center (Tsukuba, Japan). Cell line authentication of LM8 was performed by RIKEN.

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Yahiro, K., Matsumoto, Y., Yamada, H. et al. Activation of TLR4 signaling inhibits progression of osteosarcoma by stimulating CD8-positive cytotoxic lymphocytes. Cancer Immunol Immunother 69, 745–758 (2020). https://doi.org/10.1007/s00262-020-02508-9

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  • DOI: https://doi.org/10.1007/s00262-020-02508-9

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