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Identification of a novel arthritis-associated osteoclast precursor macrophage regulated by FoxM1

Nature Immunology volume 20, pages 1631–1643 (2019)Cite this article

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Abstract

Osteoclasts have a unique bone-destroying capacity, playing key roles in steady-state bone remodeling and arthritic bone erosion. Whether the osteoclasts in these different tissue settings arise from the same precursor states of monocytoid cells is presently unknown. Here, we show that osteoclasts in pannus originate exclusively from circulating bone marrow-derived cells and not from locally resident macrophages. We identify murine CX3CR1hiLy6CintF4/80+I-A+/I-E+ macrophages (termed here arthritis-associated osteoclastogenic macrophages (AtoMs)) as the osteoclast precursor-containing population in the inflamed synovium, comprising a subset distinct from conventional osteoclast precursors in homeostatic bone remodeling. Tamoxifen-inducible Foxm1 deletion suppressed the capacity of AtoMs to differentiate into osteoclasts in vitro and in vivo. Furthermore, synovial samples from human patients with rheumatoid arthritis contained CX3CR1+HLA-DRhiCD11c+CD80−CD86+ cells that corresponded to mouse AtoMs, and human osteoclastogenesis was inhibited by the FoxM1 inhibitor thiostrepton, constituting a potential target for rheumatoid arthritis treatment.

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Fig. 1: BM-derived CX3CR1+ cells differentiate into osteoclasts in pannus.
Fig. 2: CX3CR1hiLy6CintF4/80hiI-A+/I-E+ macrophages in the inflamed synovium have high osteoclast differentiation potential.
Fig. 3: The RANKL–RANK–OPG axis is essential for R3′ cell osteoclastogenesis.
Fig. 4: Transcriptional profiling by RNA-Seq identifies FoxM1 as a key regulator of R3′ cells.
Fig. 5: Single-cell RNA-Seq analysis of synovial R3 cells.
Fig. 6: M-CSF mediates R2 to R3 cell transition.
Fig. 7: FoxM1 contributes to arthritis-induced bone destruction.
Fig. 8: Rheumatoid arthritis synovial CX3CR1+HLA-DRhiCD11c+CD86+ cells have high osteoclastogenic potential.

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Data availability

The datasets analyzed during the current study are available from the corresponding author on reasonable request. Raw RNA-Seq data and single-cell RNA-Seq data related to this study are available from the Gene Expression Omnibus (accession numbers GSE117149 and GSM3712154, respectively).

Change history

  • 24 January 2020

    The caption for Source Data Extended Data Fig. 8 was given as “Gel source Data for Figure 6c”; it has been changed to “Gel source Data for Figure 7b.”

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Acknowledgements

We thank R. N. Germain (NIAID/NIH, USA) for critically reviewing this manuscript. This work was supported by CREST at the Japan Science and Technology Agency (J170701506 to M. I.), Grants-in-Aid for Scientific Research (S) from the Japan Society for the Promotion of Science (19H056570 to M.I.), a Grant-in-Aid for Young Scientists (A) from the Japan Society for the Promotion of Science (15H056710 to J.K.), and grants from the Uehara Memorial Foundation (to M.I.), Kanae Foundation for the Promotion of Medical Sciences (to M.I.), Mochida Memorial Foundation (to M.I.), Takeda Science Foundation (to M.I.) and PRIME at the Japan Agency for Medical Research and Development (19gm6210005h to J.K.).

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Authors and Affiliations

  1. Department of Immunology and Cell Biology, Graduate School of Medicine and Frontier Biosciences, Osaka University, Osaka, Japan

    Tetsuo Hasegawa, Junichi Kikuta, Takao Sudo, Yoshinobu Matsuura, Takahiro Matsui, Szandor Simmons & Masaru Ishii

  2. Division of Rheumatology, Department of Internal Medicine, Keio University School of Medicine, Tokyo, Japan

    Tetsuo Hasegawa, Kunihiro Yamaoka & Tsutomu Takeuchi

  3. WPI Immunology Frontier Research Center, Osaka University, Osaka, Japan

    Junichi Kikuta & Masaru Ishii

  4. Department of Orthopaedic Surgery, Osaka University, Osaka, Japan

    Kosuke Ebina & Makoto Hirao

  5. Genome Information Research Center, Research Institute for Microbial Diseases, Osaka University, Osaka, Japan

    Daisuke Okuzaki

  6. Department of Gastroenterology and Hepatology, Osaka University, Osaka, Japan

    Yuichi Yoshida

  7. Division of Molecular Genetics, Cancer Research Institute, WPI Nano Life Science Institute, Kanazawa University, Kanazawa, Japan

    Atsushi Hirao

  8. Center for Lung Regenerative Medicine, Division of Pulmonary Biology, Cincinnati Children’s Hospital Medical Center, Cincinnati, OH, USA

    Vladimir V. Kalinichenko

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Contributions

T.H. and M.I. conceived the study. T.H. and J.K. designed the experiments. T.S., Y.M., S.S., T.M., K.Y. and T.T. discussed the experiments and results. K.E. and M.H. provided the samples from patients with rheumatoid arthritis. Y.Y., A.H. and V.V.K. provided the Foxm1fl/fl mouse line. A.H. provided the RosaERT2Cre mouse line. D.O. performed the RNA-Seq analysis and single-cell RNA-Seq analysis. T.H. wrote the initial draft. J.K. and M.I. revised the final draft.

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Correspondence to Masaru Ishii.

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Peer review information 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.

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Supplementary information

Supplementary Information

Supplementary Figs. 1–8 and Tables 1 and 2

Reporting Summary

41590_2019_526_MOESM3_ESM.mov

Supplementary Video 1 Ex vivo incubation of inflamed synovium from double-transgenic mice (CX3CR1-EGFP and TRAP-tdTomato).

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Hasegawa, T., Kikuta, J., Sudo, T. et al. Identification of a novel arthritis-associated osteoclast precursor macrophage regulated by FoxM1. Nat Immunol 20, 1631–1643 (2019). https://doi.org/10.1038/s41590-019-0526-7

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