Abstract
Bone is the most common site of metastasis, and although low proliferation and immunoediting at the early stage make existing treatment modalities less effective, the microenvironment-inducing behaviour could be a target for early intervention. Here we report on a spatiotemporal coupling interaction between tumour cells and osteoclasts, and named the tumour-associated osteoclast ‘tumasteoclast’—a subtype of osteoclasts in bone metastases induced by tumour-migrasome-mediated cytoplasmic transfer. We subsequently propose an in situ decoupling–killing strategy in which tetracycline-modified nanoliposomes encapsulating sodium bicarbonate and sodium hydrogen phosphate are designed to specifically release high concentrations of hydrogen phosphate ions triggered by tumasteoclasts, which depletes calcium ions and forms calcium-phosphorus crystals. This can inhibit the formation of migrasomes for decoupling and disrupt cell membrane for killing, thereby achieving early prevention of bone metastasis. This study provides a research model for exploring tumour cell behaviour in detail and a proof-of-concept for behaviour-targeting strategy.
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Data availability
The main data that support the findings of this study are available within the paper and its Supplementary Information. The transcriptomic data of the OCPs, RA-OCPs, OCs and TAOCs can be found under accession no. PRJNA1045297. Further materials from this study are available from the corresponding author on reasonable request. Source Data are provided with this paper.
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Acknowledgements
This work was supported by grants from the National Nature Science Fund of China (grant nos. 82322043, 92268113 and 82072414 to XF.L., 82372454 to PF.C. and 82330077 to S.F.), the Natural Science Fund of Zhejiang Province (grant no. LGF21H060005 to Y.H.), and the Zhejiang Provincial Department of Science and Technology “Leading Geese” research and development project (grant no. 2023C03091 to S.F.). We thank H. Jiao at the School of Life Sciences of Tsinghua University for his theoretical support on migrasome formation. We thank Q. Bian and Y. Zhou at the Zhejiang Province Key Laboratory of Anti-Cancer Drug Research for their technical support on animal experiments. We thank K. Kong at the Department of Chemistry of Zhejiang University for his theoretical support on CaP crystals. We thank L. Wu, P. Yang, D. Song and G. Zhu at the Center of Cryo-Electron Microscopy (CCEM), Zhejiang University for their technical assistance on cryo-TEM, TEM and SEM.
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C.G., PF.C. and XF.L. designed the project. C.G., PF.C., H.T., Y.Y., Z.H., H.Y., K.P. and PY.C. performed the experiments. C.T., J.X. and L.S. provided technical help. C.G. and H.Y. created the visualizations. C.G., PF.C., S.F. and XF.L. wrote the manuscript. XF.L. and S.F. supervised the project. All of the authors analysed and interpreted the data, and reviewed and edited the manuscript before submission.
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XF.L., C.G. and S.F. are on a patent application (CN 2024100072569) filed by Zhejiang University related to this work. The other authors declare no competing interests.
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Nature Nanotechnology thanks Hiroki Yokota, Xiang (H.-F.) Zhang and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.
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Extended data
Extended Data Fig. 1 Schematic diagram of migrasome-mediated tumour-TAOC coupling interaction and behaviour-targeted strategy with decoupling–killing therapy by HC&HP@TNL.
Tumour cells induce spatially contacted RANKL-activated osteoclast precursor (RA-OCP) into “Tumasteoclast” (TAOC) to form tumour-TAOC spatiotemporal coupling interaction via migrasome-mediated cytoplasmic transfer. Based on the spatiotemporal characteristics of tumour-TAOC coupling in the initial metastasis, we constructed tetracycline-modified nanoliposomes encapsulating sodium bicarbonate and sodium hydrogen phosphate (HC&HP@TNL). When the bone metastasis reactivates and tumour-TAOC coupling is formed, HC&HP@TNL will be triggered by TAOC to release high concentrations of sodium hydrogen phosphate. Hydrogen phosphate combines with calcium ions in the microenvironment to form in situ calcium-phosphorus (CaP) crystals, which reduce the calcium concentration to inhibit migrasome formation, and disrupt the cell membrane integrity to induce immunogenic cell death (ICD) for immune response.
Extended Data Fig. 2 HC&HP@TNL activates the antitumour immune response.
a-c, Schematic representation of an ectopic injection model after CaP-treated 4T1 cell injection (a), distant mammary tumour image (b) and tumour volume (c) (scale bar, 10 mm, n = 8 independent mice, mean ± SD). d-f, Schematic representation of an ectopic injection model after bone metastasis treatment (d), distant mammary tumour image (e) and tumour volume (f) (scale bar, 10 mm, n = 8 independent mice, mean ± SD). g, Flow cytometric analysis of CD80 + CD86+ DCs in draining lymph nodes. h, i, Flow cytometric analysis of CD4 + T cells (h) and CD8 + T cells (i) in the distant mammary tumours (n = 6 independent mice, mean ± SD). j, HE staining and immunofluorescence images of CD8 and CD4 (scale bar, 100μm). k, Distant tumour volume in an ectopic injection model after CaP-treated 4T1 cell injection in immunocompromised mice (scale bar, 10 mm, n = 8 independent mice, mean ± SD). l, Distant tumour volume in an ectopic injection model after bone metastasis treatment in immunocompromised mice (scale bar, 10 mm, n = 4 independent mice for Cl@TNL as control, n = 8 independent mice for HC&HP@TNL, mean ± SD). m, Tumour volume of bone metastasis in BALB/c normal mice (n = 8 independent mice, mean ± SD) and immunocompromised mice (n = 4 independent mice for Cl@TNL, n = 7 independent mice for HC&HP@TNL, mean ± SD). n, Tumour inhibition rate of bone metastasis by HC&HP@TNL in normal BALB/c, immuno-enhanced BALB/c, and immunocompromised BALB/c (Cl@TNL as control of 0%, n = 8 independent mice for normal BALB/c and BALB/c+anti-PD-1, n = 7 independent mice for BALB/c-nu/nu, mean ± SD). o, Inhibition effect percent of CaP effect, ICD effect, and ICB effect in bone metastasis by HC&HP@TNL+anti-PD-1. (n = 8 independent mice, mean ± SD). P values were determined using two-way ANOVA with Šidák’s multiple-comparison test (c, f, k, l), or two-tailed one-way ANOVA with a Tukey post-hoc test (g-i, m-o).
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Supplementary Discussion 1–12, Figs. 1–16, notes and unprocessed blot images.
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Gu, C., Chen, P., Tian, H. et al. Targeting initial tumour–osteoclast spatiotemporal interaction to prevent bone metastasis. Nat. Nanotechnol. (2024). https://doi.org/10.1038/s41565-024-01613-5
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DOI: https://doi.org/10.1038/s41565-024-01613-5
