Abstract

Metastasis is the leading cause of death for cancer patients. This multi-stage process requires tumour cells to survive in the circulation, extravasate at distant sites, then proliferate; it involves contributions from both the tumour cell and tumour microenvironment (‘host’, which includes stromal cells and the immune system1). Studies suggest the early steps of the metastatic process are relatively efficient, with the post-extravasation regulation of tumour growth (‘colonization’) being critical in determining metastatic outcome2. Here we show the results of screening 810 mutant mouse lines using an in vivo assay to identify microenvironmental regulators of metastatic colonization. We identify 23 genes that, when disrupted in mouse, modify the ability of tumour cells to establish metastatic foci, with 19 of these genes not previously demonstrated to play a role in host control of metastasis. The largest reduction in pulmonary metastasis was observed in sphingosine-1-phosphate (S1P) transporter spinster homologue 2 (Spns2)-deficient mice. We demonstrate a novel outcome of S1P-mediated regulation of lymphocyte trafficking, whereby deletion of Spns2, either globally or in a lymphatic endothelial-specific manner, creates a circulating lymphopenia and a higher percentage of effector T cells and natural killer (NK) cells present in the lung. This allows for potent tumour cell killing, and an overall decreased metastatic burden.

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Acknowledgements

This work was supported by grants from Cancer Research UK (D.J.A. and O.J.S.), the Wellcome Trust (WT098051), Combat Cancer (D.J.A.), the European Research Council (311301 COLONCAN to O.J.S. and A.D.C.), National Institutes of Health U54HG004028 (N.A.K.), and Department of Defense BCRP Program Award W81XWH-14-1-0086 (S.S.). T.T. was funded by project A27N in the SFB854, and T.B. was funded in part by an EMBO Long-Term Fellowship (ALTF 945-2015) and the European Commission (Marie Curie Action LTFCOFUND2013, GA-2013-609409). We thank J. Allegood for sphingolipid analyses and acknowledge the VCU Lipidomics Core, which is supported in part by funding from the National Institutes of Health–National Cancer Institute (NIH–NCI) Cancer Center Support Grant P30CA016059, V. Iyer (Wellcome Trust Sanger Institute) for bioinformatics analysis, and members of the Wellcome Trust Sanger Institute Research Support Facility for their care of the mice.

Author information

Author notes

    • Anneliese O. Speak
    •  & David J. Adams

    These authors contributed equally to this work.

Affiliations

  1. Wellcome Trust Sanger Institute, Wellcome Genome Campus, Cambridge CB10 1SA, UK

    • Louise van der Weyden
    • , Hannah Wardle-Jones
    • , Nicola Griggs
    • , Martin Del Castillo Velasco-Herrera
    • , Natasha A. Karp
    • , Simon Clare
    • , Diane Gleeson
    • , Edward Ryder
    • , Antonella Galli
    • , Elizabeth Tuck
    • , Emma L. Cambridge
    • , Thierry Voet
    • , Iain C. Macaulay
    • , Kim Wong
    • , Anneliese O. Speak
    •  & David J. Adams
  2. University of Edinburgh Division of Pathology, Edinburgh Cancer Research UK Cancer Centre, Institute of Genetics & Molecular Medicine, Edinburgh EH4 2XR, UK

    • Mark J. Arends
  3. Cancer Research UK Beatson Institute, Glasgow G61 1BD, UK

    • Andrew D. Campbell
    •  & Owen J. Sansom
  4. Department of Dermatology, University Hospital Magdeburg, Magdeburg 39120, Germany

    • Tobias Bald
    •  & Thomas Tüting
  5. Department of Immunology in Cancer and Infection Laboratory, QIMR Berghofer Medical Research Institute, Herston 4006, Australia

    • Tobias Bald
  6. Department of Human Genetics, University of Leuven (KU Leuven), Leuven, 3000, Belgium

    • Thierry Voet
  7. Department of Biochemistry and Molecular Biology, Virginia Commonwealth University School of Medicine, Richmond, Virginia 23298-0614, USA

    • Sarah Spiegel

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  1. Sanger Mouse Genetics Project

    Lists of participants and their affiliations appear in the Supplementary Information

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Contributions

L.v.d.W. devised and implemented the pulmonary metastasis screen, performing all the primary screen, confirmation and characterization studies. M.J.A. analysed the histopathological sections. A.D.C. and O.J.S. performed and analysed the intrasplenic B16-F10 assays. T.B. and T.T. performed and analysed the spontaneous metastasis assay. H.W.-J. and N.G. managed mouse breeding and were responsible for issuing phenotyping cohorts. M.D.C.V.-H., T.V., I.C.M. and K.W. performed the RNA-seq analysis. D.G. and E.R. genotyped the mice and performed gene expression analysis. S.C., A.G., E.T. and E.L.C. performed additional phenotypic characterization. The Sanger Mouse Genetics Project generated and phenotyped the mice as part of a primary phenotyping pipeline. S.S. oversaw the lipidomic analysis and provided input to the project and the manuscript. A.O.S. devised, performed and analysed the immunophenotyping assays. L.v.d.W., A.O.S. and D.J.A. led the project. L.v.d.W., A.O.S. and D.J.A. wrote the manuscript with contributions from all authors.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Louise van der Weyden or David J. Adams.

Reviewer Information: Nature thanks C. Ghajar and the other anonymous reviewer(s) for their contribution to the peer review of this work.

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    Supplementary Information

    This file contains a list of the Sanger Mouse Genetics Project participants and Supplementary Tables 1-4.

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DOI

https://doi.org/10.1038/nature20792

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