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The perivascular niche regulates breast tumour dormancy

Nature Cell Biology volume 15, pages 807–817 (2013)Cite this article

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Abstract

In a significant fraction of breast cancer patients, distant metastases emerge after years or even decades of latency. How disseminated tumour cells (DTCs) are kept dormant, and what wakes them up, are fundamental problems in tumour biology. To address these questions, we used metastasis assays in mice and showed that dormant DTCs reside on microvasculature of lung, bone marrow and brain. We then engineered organotypic microvascular niches to determine whether endothelial cells directly influence breast cancer cell (BCC) growth. These models demonstrated that endothelial-derived thrombospondin-1 induces sustained BCC quiescence. This suppressive cue was lost in sprouting neovasculature; time-lapse analysis showed that sprouting vessels not only permit, but accelerate BCC outgrowth. We confirmed this surprising result in dormancy models and in zebrafish, and identified active TGF-β1 and periostin as tumour-promoting factors derived from endothelial tip cells. Our work reveals that stable microvasculature constitutes a dormant niche, whereas sprouting neovasculature sparks micrometastatic outgrowth.

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Figure 1: Dormant breast tumour cells reside on microvascular endothelium in distant tissues in vivo.
Figure 2: Microvascular endothelium induces sustained quiescence of breast tumour cells in engineered cultures.
Figure 3: TSP-1 is an angiocrine tumour suppressor.
Figure 4: Opposite regulation of tumour dormancy and growth by endothelial sub-niches: stable endothelium inhibits—whereas neovascular tips promote—breast tumour cell growth.
Figure 5: Notch1-mediated reduction in neovascular tips suppresses breast tumour cell outgrowth.
Figure 6: Ectopic vascular sprouting promotes growth of injected breast tumour cells in zebrafish larvae.
Figure 7: Neovascular tips comprise micrometastatic niches enriched for POSTN and TGF-β1.

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Acknowledgements

We thank S. Rafii and B. Weinstein for generously providing the E4ORF1 lentiviral plasmid and stalactite mutant zebrafish, respectively. We are grateful to N. Boudreau, S. Rafii, R. Schwartz, R. Xu, A. Bruni-Cardoso and A.L. Correia for critical insight, and to other present members of the Bissell laboratory for helpful discussions. We thank A. Lo and C. Williams for technical assistance and T. Varimezova for performing blinded quantitative analysis. C.M.G. was financially supported by a Glenn T. Seaborg Postdoctoral Fellowship from LBNL and by a NCI-U54CA143836 training grant. K.J.E. is a Robert Black Fellow supported by the Damon Runyon Cancer Research Foundation (DRG-109-10). K.A.H.’s laboratory is supported by grants from the NIH (R01HL042493 and R01HL090895) and from the March of Dimes Foundation (6-FY12-356). The work in D.Y.R.S.’s laboratory was financially supported in part by grants from the NIH (HL54737) and the Packard Foundation. E.I.C.’s laboratory is supported by a Carol Baldwin Breast Cancer Award, a shared instrument grant (NIH/NCRR 1 S10 RR023680-1) and a DOE Subcontract (DE-AC02-05CH1123) from LBNL. D.L. is supported by The Hartwell Foundation. The work from M.J.B.’s laboratory is supported by grants from the US Department of Energy, Office of Biological and Environmental Research and Low Dose Scientific Focus Area (contract no. DE-AC02-05CH1123); by the National Cancer Institute (awards U01CA169538 (to D.L. and M.J.B.), U54CA126552, R37CA064786 and U54CA143836—Bay Area Physical Sciences–Oncology Center, University of California, Berkeley, California); by a grant from the Breast Cancer Research Foundation; and by the US Department of Defense (W81XWH0810736).

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Author notes

  1. Didier Y. R. Stainier

    Present address: Present address: Department of Developmental Genetics, Max Planck Institute for Heart and Lung Research, Bad Nauheim D-61231, Germany,

Authors and Affiliations

  1. Life Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, California 94720, USA

    Cyrus M. Ghajar, Hidetoshi Mori & Mina J. Bissell

  2. Department of Pediatrics, Weill Cornell Medical College, New York 10021, USA

    Héctor Peinado, Irina R. Matei, Hélène Brazier, Katherine A. Hajjar & David Lyden

  3. Department of Cell and Developmental Biology, Weill Cornell Medical College, New York 10065, USA

    Héctor Peinado, Irina R. Matei, Hélène Brazier, Dena Almeida, Katherine A. Hajjar & David Lyden

  4. Department of Pathology, University of California, San Francisco, California 94143, USA

    Kimberley J. Evason

  5. Stony Brook University Proteomics Center, School of Medicine, Stony Brook, New York 11794, USA

    Antonius Koller & Emily I. Chen

  6. Department of Biochemistry and Biophysics, University of California, San Francisco, California 94158, USA

    Kimberley J. Evason & Didier Y. R. Stainier

  7. Department of Pharmacological Sciences, Stony Brook University, Stony Brook, New York 11794, USA

    Emily I. Chen

  8. Department of Pediatrics, Memorial Sloan-Kettering Cancer Center, New York 10065, USA

    David Lyden

  9. Champalimaud Metastasis Programme, Lisbon 1400-038, Portugal

    David Lyden

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Contributions

C.M.G., D.L. and M.J.B. conceived of the study. C.M.G., H.P., I.R.M. and H.B. conducted animal studies and analysed resulting data. C.M.G., K.J.E. and D.Y.R.S. planned and executed zebrafish experiments. D.A. and K.A.H. lent expertise in the retinal neovascularization model. C.M.G. and H.M. collected and analysed data, and H.M. also provided conceptual advice. A.K. and E.I.C. conducted LC–MS/MS analysis and analysed data. C.M.G. and M.J.B. wrote the manuscript. All authors read and critiqued the manuscript extensively.

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Correspondence to Cyrus M. Ghajar or Mina J. Bissell.

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Supplementary information

Supplementary Information

Supplementary Information (PDF 1466 kb)

Opposite regulation of tumour quiescence and growth by endothelial sub-niches: stable endothelium inhibits breast tumour cell growth.

H2B-GFP T4-2 cells residing predominantly around stable endothelium does not undergo division over a 72 h period. Intervals, 20 min. Scale bar, 50 μm. (MOV 4280 kb)

Opposite regulation of tumour quiescence and growth by endothelial sub-niches: neovascular tips promote breast tumour cell growth.

H2B-GFP T4-2 cells are observed dividing soon after encountering neovascular tips. Highlighted cell divides after encountering an endothelial ‘probe’. At the right of the screen, another cell divides at 1 d, 16:00, as it encounters an approaching neovascular tip. One of the daughter cells divides again as the couplet resumes interaction with the tip. At the bottom, another cell ‘surfs’ past a neovascular tip and eventually divides. Intervals, 20 min. Scale bar, 100 μm. (MOV 4219 kb)

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Ghajar, C., Peinado, H., Mori, H. et al. The perivascular niche regulates breast tumour dormancy. Nat Cell Biol 15, 807–817 (2013). https://doi.org/10.1038/ncb2767

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