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VEGFR1-positive haematopoietic bone marrow progenitors initiate the pre-metastatic niche

Abstract

The cellular and molecular mechanisms by which a tumour cell undergoes metastasis to a predetermined location are largely unknown. Here we demonstrate that bone marrow-derived haematopoietic progenitor cells that express vascular endothelial growth factor receptor 1 (VEGFR1; also known as Flt1) home to tumour-specific pre-metastatic sites and form cellular clusters before the arrival of tumour cells. Preventing VEGFR1 function using antibodies or by the removal of VEGFR1+ cells from the bone marrow of wild-type mice abrogates the formation of these pre-metastatic clusters and prevents tumour metastasis, whereas reconstitution with selected Id3 (inhibitor of differentiation 3)-competent VEGFR1+ cells establishes cluster formation and tumour metastasis in Id3 knockout mice. We also show that VEGFR1+ cells express VLA-4 (also known as integrin α4β1), and that tumour-specific growth factors upregulate fibronectin—a VLA-4 ligand—in resident fibroblasts, providing a permissive niche for incoming tumour cells. Conditioned media obtained from distinct tumour types with unique patterns of metastatic spread redirected fibronectin expression and cluster formation, thereby transforming the metastatic profile. These findings demonstrate a requirement for VEGFR1+ haematopoietic progenitors in the regulation of metastasis, and suggest that expression patterns of fibronectin and VEGFR1+VLA-4+ clusters dictate organ-specific tumour spread.

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Acknowledgements

We thank M. Barna for critical reading of the manuscript and L. Breda, S. Rivella and S. Neustein for discussions. R.N.K. is a recipient of the Laura Rosenberg Fellowship award and supported by a grant from the American Hellenic Educational Progressive Association (Fifth District) and the LTC Foundation. D.L. is supported by the Doris Duke Charitable Foundation, the Children's Blood Foundation, the Emerald Foundation, the Theodore A. Rapp Foundation and a grant from the National Cancer Institute. S.R. is an investigator of the Howard Hughes Medical Institute and supported by grants from the American Cancer Society, the Leukemia and Lymphoma Society, and the National Institutes of Health.

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Reprints and permissions information is available at npg.nature.com/reprintsandpermissions. The authors declare no competing financial interests.

Correspondence to Shahin Rafii or David Lyden.

Supplementary information

Supplementary Figure 1

Tumour and VEGFR2+ cells arrive after VEGFR1+ HPCs. (PDF 139 kb)

Supplementary Figure 2

VEGFR1 antibody inhibits metastasis in B16 tumours. (PDF 129 kb)

Supplementary Figure 3

Inhibition of VLA-4 and MMP-9 inhibits metastasis. B16 melanoma induces fibronectin expression in the lung. (PDF 128 kb)

Supplementary Figure 4

Bone marrow-derived VEGFR1+ cells attract tumour cells. (PDF 159 kb)

Supplemental Figure 5

Tumours vary in their chemokine profile and fibronectin expression pattern. (PDF 101 kb)

Supplementary Table 1

Id3 deficient mice have decreased mobilization of CD11b+ VEGFR1+ hematopoietic progenitors. (PDF 33 kb)

Supplementary Methods

Additional details of the methods used in this study. (DOC 35 kb)

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Further reading

Figure 1: Bone marrow-derived cells form the pre-metastatic niche.
Figure 2: Pre-metastatic clusters are comprised of VEGFR1 + haematopoietic progenitors.
Figure 3: Expression of VEGFR1 in pre-metastatic human tissue.
Figure 4: Inhibition of homing of bone marrow cells prevents metastasis.
Figure 5: The VLA-4/fibronectin pathway mediates cluster formation.
Figure 6: Redirection of LLC metastases to atypical sites.

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