Abstract

Ever since Stephen Paget’s 1889 hypothesis, metastatic organotropism has remained one of cancer’s greatest mysteries. Here we demonstrate that exosomes from mouse and human lung-, liver- and brain-tropic tumour cells fuse preferentially with resident cells at their predicted destination, namely lung fibroblasts and epithelial cells, liver Kupffer cells and brain endothelial cells. We show that tumour-derived exosomes uptaken by organ-specific cells prepare the pre-metastatic niche. Treatment with exosomes from lung-tropic models redirected the metastasis of bone-tropic tumour cells. Exosome proteomics revealed distinct integrin expression patterns, in which the exosomal integrins α6β4 and α6β1 were associated with lung metastasis, while exosomal integrin αvβ5 was linked to liver metastasis. Targeting the integrins α6β4 and αvβ5 decreased exosome uptake, as well as lung and liver metastasis, respectively. We demonstrate that exosome integrin uptake by resident cells activates Src phosphorylation and pro-inflammatory S100 gene expression. Finally, our clinical data indicate that exosomal integrins could be used to predict organ-specific metastasis.

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Gene Expression Omnibus

Data deposits

The raw data for quantitative mass spectrometry analysis of lung-tropic (4173 and 4175), liver-tropic (HPAF-II and HCT116) and brain-tropic (831 and 231BR) exosomes (Fig. 2a and Extended Data Fig. 3a) are available at http://dx.doi.org/10.6084/m9.figshare.1569781. The raw sequencing data for human Kupffer cells treated in vitro with BxPC-3 or BxPC-3 ITGβ5KD exosomes have been deposited in the Gene Expression Omnibus (GEO) under accession number GSE68919.

References

  1. 1.

    The distribution of secondary growths in cancer of the breast. 1889. Cancer Metastasis Rev. 8, 98–101 (1989)

  2. 2.

    & Role of organ selectivity in the determination of metastatic patterns of B16 melanoma. Cancer Res. 40, 2281–2287 (1980)

  3. 3.

    et al. Involvement of chemokine receptors in breast cancer metastasis. Nature 410, 50–56 (2001)

  4. 4.

    , & Cancer to bone: a fatal attraction. Nature Rev. Cancer 11, 411–425 (2011)

  5. 5.

    et al. Cancer-secreted miR-105 destroys vascular endothelial barriers to promote metastasis. Cancer Cell 25, 501–515 (2014)

  6. 6.

    et al. The IL-6/JAK/Stat3 feed-forward loop drives tumorigenesis and metastasis. Neoplasia 15, 848–862 (2013)

  7. 7.

    & Organotropism of breast cancer metastasis. J. Mammary Gland Biol. Neoplasia 12, 153–162 (2007)

  8. 8.

    et al. The hypoxic cancer secretome induces pre-metastatic bone lesions through lysyl oxidase. Nature 522, 106–110 (2015)

  9. 9.

    et al. VEGFR1-positive haematopoietic bone marrow progenitors initiate the pre-metastatic niche. Nature 438, 820–827 (2005)

  10. 10.

    et al. MMP9 induction by vascular endothelial growth factor receptor-1 is involved in lung-specific metastasis. Cancer Cell 2, 289–300 (2002)

  11. 11.

    et al. Melanoma exosomes educate bone marrow progenitor cells toward a pro-metastatic phenotype through MET. Nature Med. 18, 883–891 (2012)

  12. 12.

    et al. Tumour microvesicles contain retrotransposon elements and amplified oncogene sequences. Nature Commun. 2, 180 (2011)

  13. 13.

    et al. Glioblastoma microvesicles transport RNA and proteins that promote tumour growth and provide diagnostic biomarkers. Nature Cell Biol. 10, 1470–1476 (2008)

  14. 14.

    , & Membrane vesicles as conveyors of immune responses. Nature Rev. Immunol. 9, 581–593 (2009)

  15. 15.

    & Extracellular vesicles: exosomes, microvesicles, and friends. J. Cell Biol. 200, 373–383 (2013)

  16. 16.

    , & The secreted factors responsible for pre-metastatic niche formation: old sayings and new thoughts. Semin. Cancer Biol. 21, 139–146 (2011)

  17. 17.

    , , & Proteomics, transcriptomics and lipidomics of exosomes and ectosomes. Proteomics 13, 1554–1571 (2013)

  18. 18.

    et al. Exosome-mediated transfer of mRNAs and microRNAs is a novel mechanism of genetic exchange between cells. Nature Cell Biol. 9, 654–659 (2007)

  19. 19.

    et al. Double-stranded DNA in exosomes: a novel biomarker in cancer detection. Cell Res. 24, 766–769 (2014)

  20. 20.

    et al. Pancreatic cancer exosomes initiate pre-metastatic niche formation in the liver. Nature Cell Biol. 17, 816–826 (2015)

  21. 21.

    et al. A multigenic program mediating breast cancer metastasis to bone. Cancer Cell 3, 537–549 (2003)

  22. 22.

    et al. Identifying site-specific metastasis genes and functions. Cold Spring Harb. Symp. Quant. Biol. 70, 149–158 (2005)

  23. 23.

    et al. Genes that mediate breast cancer metastasis to lung. Nature 436, 518–524 (2005)

  24. 24.

    et al. Genes that mediate breast cancer metastasis to the brain. Nature 459, 1005–1009 (2009)

  25. 25.

    & Integrins in cancer: biological implications and therapeutic opportunities. Nature Rev. Cancer 10, 9–22 (2010)

  26. 26.

    , , , & The minimum element of a synthetic peptide required to block prostate tumor cell migration. Cancer Biol. Ther. 5, 1556–1562 (2006)

  27. 27.

    & Arg-Gly-Asp: a versatile cell recognition signal. Cell 44, 517–518 (1986)

  28. 28.

    et al. Suppression of tumor development and metastasis formation in mice lacking the S100A4(mts1) gene. Cancer Res. 65, 3772–3780 (2005)

  29. 29.

    & Building the niche: the role of the S100 proteins in metastatic growth. Semin. Cancer Biol. 22, 216–225 (2012)

  30. 30.

    , , , & Integrin (α6β4) signals through Src to increase expression of S100A4, a metastasis-promoting factor: implications for cancer cell invasion. Mol. Cancer Res. 7, 1605–1612 (2009)

  31. 31.

    , , & Focal adhesion kinase activated by β4 integrin ligation to mCLCA1 mediates early metastatic growth. J. Biol. Chem. 277, 34391–34400 (2002)

  32. 32.

    et al. p38 MAPK is a critical regulator of the constitutive and the beta4 integrin-regulated expression of IL-6 in human normal thymic epithelial cells. Eur. J. Immunol. 33, 3038–3048 (2003)

  33. 33.

    et al. β4 integrin-dependent formation of polarized three-dimensional architecture confers resistance to apoptosis in normal and malignant mammary epithelium. Cancer Cell 2, 205–216 (2002)

  34. 34.

    et al. Targeted deletion of the integrin beta4 signaling domain suppresses laminin-5-dependent nuclear entry of mitogen-activated protein kinases and NF-κB, causing defects in epidermal growth and migration. Mol. Cell. Biol. 25, 6090–6102 (2005)

  35. 35.

    et al. Distinct organ-specific metastatic potential of individual breast cancer cells and primary tumors. J. Clin. Invest. 115, 44–55 (2005)

  36. 36.

    et al. Breast cancer cells produce tenascin C as a metastatic niche component to colonize the lungs. Nature Med. 17, 867–874 (2011)

  37. 37.

    et al. Periostin is a key niche component for wound metastasis of melanoma. PLoS ONE 10, e0129704 (2015)

  38. 38.

    , & Order and disorder: the role of extracellular matrix in epithelial cancer. Cancer Invest. 20, 139–153 (2002)

  39. 39.

    et al. Reversion of the malignant phenotype of human breast cells in three-dimensional culture and in vivo by integrin blocking antibodies. J. Cell Biol. 137, 231–245 (1997)

  40. 40.

    et al. Lung metastasis fails in MMTV-PyMT oncomice lacking S100A4 due to a T-cell deficiency in primary tumors. Cancer Res. 70, 936–947 (2010)

  41. 41.

    , , , & Integrin α6β4 controls the expression of genes associated with cell motility, invasion, and metastasis, including S100A4/metastasin. J. Biol. Chem. 284, 1484–1494 (2009)

  42. 42.

    et al. Cilengitide inhibits progression of experimental breast cancer bone metastases as imaged noninvasively using VCT, MRI and DCE-MRI in a longitudinal in vivo study. Int. J. Cancer 128, 2453–2462 (2011)

  43. 43.

    et al. Targeting αV-integrins decreased metastasis and increased survival in a nude rat breast cancer brain metastasis model. J. Neurooncol. 110, 27–36 (2012)

  44. 44.

    et al. Tumor αvβ3 integrin is a therapeutic target for breast cancer bone metastases. Cancer Res. 67, 5821–5830 (2007)

  45. 45.

    et al. Brain metastatic cancer cells release microRNA-181c-containing extracellular vesicles capable of destructing blood-brain barrier. Nature Commun. 6, 6716 (2015)

  46. 46.

    et al. Melanoma exosomes educate bone marrow progenitor cells toward a pro-metastatic phenotype through MET. Nature Med. 18, 883–891 (2012)

  47. 47.

    , & Stop and go extraction tips for matrix-assisted laser desorption/ionization, nanoelectrospray, and LC/MS sample pretreatment in proteomics. Anal. Chem. 75, 663–670 (2003)

  48. 48.

    , , , & The minotaur proteome: avoiding cross-species identifications deriving from bovine serum in cell culture models. Proteomics 10, 3040–3044 (2010)

  49. 49.

    , , , & Semi-supervised learning for peptide identification from shotgun proteomics datasets. Nature Methods 4, 923–925 (2007)

  50. 50.

    , , , & Absolute quantification of proteins by LCMSE: a virtue of parallel MS acquisition. Mol. Cell. Proteomics 5, 144–156 (2006)

  51. 51.

    et al. Accurate proteome-wide label-free quantification by delayed normalization and maximal peptide ratio extraction, termed MaxLFQ. Mol. Cell. Proteomics 13, 2513–2526 (2014)

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Acknowledgements

We thank S. Rudchenko at the Hospital for Special Surgery Flow Cytometry Core Facility. We acknowledge the MSK Cancer Center Support Grant/Core Grant (P30 CA008748). Our work is supported by grants from National Cancer Institute (U01-CA169538, D.L. and M.S.B.), National Institutes of Health (R01-CA169416, D.L. and H.P.), United States Department of Defense (W81XWH-13-10249, D.L.), W81XWH-13-1-0425 (D.L., J.B., B.A.G. and Y.K.), Melanoma Research Alliance (H.P.), Sohn Conference Foundation (H.P. and H.Z.), the Children’s Cancer and Blood Foundation (H.P. and D.L.), The Manning Foundation (D.L.), The Hartwell Foundation (D.L.), Fundação para a Ciência e a Técnologia (D.L.), The Nancy C. and Daniel P. Paduano Foundation (H.P. and D.L.), The Feldstein Foundation (H.P.), The Starr Cancer Consortium (H.P. and D.L.), The Mary Kay Foundation (D.L.), Pediatric Oncology Experimental Therapeutic Investigator Consortium (POETIC, D.L. and H.P.), James Paduano Foundation (D.L. and H.P.), Beth Tortolani Foundation (D.L. and J.B.), Malcolm Hewitt Weiner Foundation (D.L.), Theodore A. Rapp Foundation (D.L.), American Hellenic Educational Progressive Association 5th District Cancer Research Foundation (D.L., A.H.), Charles and Marjorie Holloway Foundation (J.B.), Sussman Family Fund (J.B.), Lerner Foundation (J.B.), Breast Cancer Alliance (J.B.), Manhasset Women’s Coalition Against Breast Cancer (J.B.), Ministry of Science and Technology Taiwan (101-2918-I-002-016, T.-L.S.), The JSPS Postdoctoral Fellowships for Research Abroad and Susan G. Komen Postdoctoral Fellowship (A.H.).

Author information

Author notes

    • Ayuko Hoshino
    • , Bruno Costa-Silva
    •  & Tang-Long Shen

    These authors contributed equally to this work.

Affiliations

  1. Children’s Cancer and Blood Foundation Laboratories, Departments of Pediatrics, and Cell and Developmental Biology, Drukier Institute for Children’s Health, Meyer Cancer Center, Weill Cornell Medicine, New York, New York 10021, USA

    • Ayuko Hoshino
    • , Bruno Costa-Silva
    • , Tang-Long Shen
    • , Goncalo Rodrigues
    • , Ayako Hashimoto
    • , Angela Di Giannatale
    • , Swarnima Singh
    • , Caitlin Williams
    • , Linda Bojmar
    • , Yonathan Ararso
    • , Haiying Zhang
    • , Jonathan Hernandez
    • , Joshua M. Weiss
    • , Maria de Sousa
    • , Mary S. Brady
    • , Irina Matei
    • , Hector Peinado
    •  & David Lyden
  2. Department of Plant Pathology and Microbiology and Center for Biotechnology, National Taiwan University, Taipei 10617, Taiwan

    • Tang-Long Shen
  3. Graduate Program in Areas of Basic and Applied Biology, Abel Salazar Biomedical Sciences Institute, University of Porto, 4099-003 Porto, Portugal

    • Goncalo Rodrigues
    •  & Maria de Sousa
  4. Department of Obstetrics and Gynecology, Faculty of Medicine, University of Tokyo, Tokyo 113-8655, Japan

    • Ayako Hashimoto
  5. Proteomics Resource Center, The Rockefeller University, New York, New York 10065, USA

    • Milica Tesic Mark
    •  & Henrik Molina
  6. Department of Pathology, Memorial Sloan Kettering Cancer Center, New York, New York 10065, USA

    • Shinji Kohsaka
  7. Department of Oncology and Pathology, Karolinska Institutet, 17176 Stockholm, Sweden

    • Sophia Ceder
  8. Electron Microscopy Resource Center (EMRC), Rockefeller University, New York, New York 10065, USA

    • Nadine Soplop
    •  & Kunihiro Uryu
  9. Breast Service, Department of Surgery, Memorial Sloan Kettering Cancer Center, New York, New York, 10065, USA

    • Lindsay Pharmer
    •  & Tari King
  10. Department of Surgery, County Council of Östergötland, and Department of Clinical and Experimental Medicine, Faculty of Health Sciences, Linköping University, 58185 Linköping, Sweden

    • Linda Bojmar
    •  & Per Sandstrom
  11. Life Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, California 94720, USA

    • Alexander E. Davies
    •  & Mina J. Bissell
  12. Genomics Resources Core Facility, Weill Cornell Medicine, New York, New York 10021, USA

    • Tuo Zhang
  13. Department of Surgery, Memorial Sloan Kettering Cancer Center, New York, New York 10065, USA

    • Jonathan Hernandez
    •  & William R. Jarnagin
  14. Department of Pediatrics, Memorial Sloan Kettering Cancer Center, New York, New York 10065, USA

    • Vanessa D. Dumont-Cole
    • , Kimberly Kramer
    • , Leonard H. Wexler
    •  & David Lyden
  15. Division of Pediatric Oncology, Alberta Children’s Hospital, Calgary, Alberta T3B 6A8, Canada

    • Aru Narendran
  16. Division of Hematology/Oncology, Columbia University School of Medicine, New York, New York 10032, USA

    • Gary K. Schwartz
  17. Orthopaedic Service, Department of Surgery, Memorial Sloan Kettering Cancer Center, New York, New York 10065, USA

    • John H. Healey
  18. Department of Hepato-Pancreato-Biliary Surgery, Oslo University Hospital, Nydalen, Oslo 0424, Norway

    • Knut Jørgen Labori
  19. Department of Cancer Genetics, Institute for Cancer Research, Oslo University Hospital, Nydalen, Oslo 0424, Norway

    • Elin H. Kure
  20. Eppley Institute for Research in Cancer and Allied Diseases, University of Nebraska Medical Center, Omaha, Nebraska 68198, USA

    • Paul M. Grandgenett
    •  & Michael A. Hollingsworth
  21. Department of Biochemistry and Molecular Biology, University of Nebraska Medical Center, Omaha, Nebraska 68198, USA

    • Sukhwinder Kaur
    • , Maneesh Jain
    • , Kavita Mallya
    •  & Surinder K. Batra
  22. Gastric and Mixed Tumor Service, Department of Surgery, Memorial Sloan Kettering Cancer Center, New York, New York 10065, USA

    • Mary S. Brady
  23. Department of Tumor Biology, Norwegian Radium Hospital, Oslo University Hospital, Nydalen, Oslo 0424, Norway

    • Oystein Fodstad
  24. Institute for Clinical Medicine, Faculty of Medicine, University of Oslo, Blindern, Oslo 0318, Norway

    • Oystein Fodstad
  25. Department of Gynecology, University Medical Center, Martinistrasse 52, 20246 Hamburg, Germany

    • Volkmar Muller
  26. Department of Tumor Biology, University Medical Center Hamburg-Eppendorf, Martinistrasse 52, 20246 Hamburg, Germany

    • Klaus Pantel
  27. Department of Radiation Oncology, Abramson Family Cancer Research Institute, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA

    • Andy J. Minn
  28. Department of Biochemistry and Biophysics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA

    • Benjamin A. Garcia
  29. Department of Molecular Biology, Princeton University, Princeton, New Jersey 08544, USA

    • Yibin Kang
  30. Rutgers Cancer Institute of New Jersey, New Brunswick, New Jersey 08903, USA

    • Yibin Kang
  31. Breast Medicine Service, Department of Medicine, Memorial Sloan Kettering Cancer Center, New York, New York 10065, USA

    • Vinagolu K. Rajasekhar
  32. Fred Hutchinson Cancer Research Center, Seattle, Washington 98109, USA

    • Cyrus M. Ghajar
  33. Microenvironment and Metastasis Laboratory, Department of Molecular Oncology, Spanish National Cancer Research Center (CNIO), Madrid 28029, Spain

    • Hector Peinado
  34. Department of Medicine, Memorial Sloan Kettering Cancer Center, New York, New York 10065, USA

    • Jacqueline Bromberg
  35. Department of Medicine, Weill Cornell Medicine, New York, New York 10021, USA

    • Jacqueline Bromberg

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Contributions

A.H. designed the experimental approach, performed the experimental work, analysed the data, coordinated the project and wrote the manuscript. B.C.-S. designed experiments investigating liver metastasis and performed the experimental work. T.-L.S. performed ECM studies. G.R. analysed brain tropic exosome distribution. A.H. performed western blot analysis. M.T.M. and H.M. performed and analysed exosome mass spectrometry. S.K. prepared overexpression vectors. S.S. and L.B. performed tissue processing and staining. S.C. designed and illustrated Fig. 4c. A.D.G., S.C., V.D.D.-C., Y.A. and C.W. received and processed human samples. N.S. and K.U. performed electron microscopy. A.E.D. performed animal surgeries and contributed to data interpretation and discussion. T.Z. performed RNA sequence analysis. B.A.G. performed initial proteomic analysis. V.K.R., G.K.S. and J.H.H. provided the uveal melanoma cell line. L.P., T.K., M.S.B., V.M., K.K., L.H.W., J.H., E.H.K., K.M., S.K.B., K.P., O.F., M.J., S.K., M.A.H., P.M.G., K.J.L., J.M.W., A.N. and W.R.J. provided and prepared human samples. H.Z., A.J.M. and P.S. read the manuscript and provided feedback. C.M.G., I.M. and H.P. discussed the hypothesis and contributed to data interpretation and wrote the manuscript. Y.K., M.d.S. and M.J.B. contributed to discussing the hypothesis, interpretation of data. J.B. coordinated the project, interpreted data and wrote the manuscript. D.L. conceived the hypothesis, led the project, interpreted the data and wrote the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Hector Peinado or Jacqueline Bromberg or David Lyden.

Extended data

Supplementary information

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  1. 1.

    Supplementary Figure

    This file contains western blot source data. a-h, Analysis of ITGβ4, α6, β1, α2, β3, αV, β5 and α3 expression, respectively, in organotropic cell line-derived exosomes in Fig. 2b. i-k, Analysis of ITGβ4, ITGα6 and GAPDH expression, respectively, in cell lysates versus exosomes derived from organotropic breast cancer and pancreatic cancer cell lines in Extended Fig. 3c. l, Analysis of ITGβ4 and GAPDH expression in 4175-LuT and 4175β4KD cells and exosomes in Extended Fig. 6a. m, Analysis of ITGβ4 and GAPDH expression in exosomes isolated from WT or ITGβ4-overexpressing1833-BoT cells in Extended Fig. 6c. n, Analysis of ITGβ5 and GAPDH expression in BxPC-3 and BxPC-3β5KD cells and exosomes in Extended Fig. 6e. o, Exosomal ITGβ4 and Alix levels in the plasma of mice bearing orthotopic 4175-Lu-T tumors in Extended Fig. 8a. LC, loading control.

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https://doi.org/10.1038/nature15756

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