Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Letter
  • Published:

In vivo imaging of specialized bone marrow endothelial microdomains for tumour engraftment

Nature volume 435, pages 969–973 (2005)Cite this article

Abstract

The organization of cellular niches is known to have a key role in regulating normal stem cell differentiation and regeneration, but relatively little is known about the architecture of microenvironments that support malignant metastasis1,2. Using dynamic in vivo confocal imaging, here we show that murine bone marrow contains unique anatomic regions defined by specialized endothelium. This vasculature expresses the adhesion molecule E-selectin and the chemoattractant stromal-cell-derived factor 1 (SDF-1) in discrete, discontinuous areas that influence the homing of a variety of tumour cell lines. Disruption of the interactions between SDF-1 and its receptor CXCR4 inhibits the homing of Nalm-6 cells (an acute lymphoblastic leukaemia cell line) to these vessels. Further studies revealed that circulating leukaemic cells can engraft around these vessels, suggesting that this molecularly distinct vasculature demarcates a microenvironment for early metastatic tumour spread in bone marrow. Finally, purified haematopoietic stem/progenitor cells and lymphocytes also localize to the same microdomains, indicating that this vasculature might also function in benign states to demarcate specific portals for the entry of cells into the marrow space. Specialized vascular structures therefore appear to delineate a microenvironment with unique physiology that can be exploited by circulating malignant cells.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Leukaemic cell homing and engraftment in mouse skull bone marrow in vivo.
Figure 2: In vivo immunofluorescence microscopy of vascular cell adhesion molecule expression in bone marrow.
Figure 3: Leukaemic cell homing to SDF-1 + E-selectin + microvascular domains is inhibited by SDF-1/CXCR4 blockade.
Figure 4: HSPCs home to SDF-1-positive vascular microdomains.

Similar content being viewed by others

References

  1. Fuchs, E., Tumbar, T. & Guasch, G. Socializing with the neighbors: Stem cells and their niche. Cell 116, 769–778 (2004)

    Article  CAS  PubMed  Google Scholar 

  2. Chambers, A. F., Groom, A. C. & MacDonald, I. C. Dissemination and growth of cancer cells in metastatic sites. Nature Rev. Cancer 2, 563–572 (2002)

    Article  CAS  Google Scholar 

  3. Honn, K. V. & Tang, D. G. Adhesion molecules and tumor cell interaction with endothelium and endothelial matrix. Cancer Metastasis Rev. 11, 353–375 (1992)

    Article  CAS  PubMed  Google Scholar 

  4. Burger, J. A. & Kipps, T. J. Chemokine receptors and stromal cells in the homing and homeostasis of chronic lymphocytic leukemia B cells. Leuk. Lymphoma 43, 461–466 (2002)

    Article  CAS  PubMed  Google Scholar 

  5. Reuss-Borst, M. A., Klein, G., Waller, H. D. & Muller, C. A. Differential expression of adhesion molecules in acute leukemia. Leukemia 9, 869–874 (1995)

    CAS  PubMed  Google Scholar 

  6. Mazo, I. B. et al. Hematopoietic progenitor cell rolling in bone marrow microvessels: Parallel contributions by endothelial selectins and vascular cell adhesion molecule 1. J. Exp. Med. 188, 465–474 (1998)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Cristy, M. Active bone marrow distribution as a function of age in humans. Phys. Med. Biol. 26, 389–400 (1981)

    Article  CAS  PubMed  Google Scholar 

  8. Butcher, E. C. Leukocyte–endothelial cell recognition: three (or more) steps to specificity and diversity. Cell 67, 1033–1036 (1991)

    Article  CAS  PubMed  Google Scholar 

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

    Article  ADS  PubMed  Google Scholar 

  10. Murphy, P. M. Chemokines and the molecular basis of cancer metastasis. N. Engl. J. Med. 345, 833–835 (2001)

    Article  CAS  PubMed  Google Scholar 

  11. Shen, W., Bendall, L. J., Gottlieb, D. J. & Bradstock, K. F. The chemokine receptor CXCR4 enhances integrin-mediated in vitro adhesion and facilitates engraftment of leukemic precursor-B cells in the bone marrow. Exp. Hematol. 29, 1439–1447 (2001)

    Article  CAS  PubMed  Google Scholar 

  12. Tavor, S. et al. CXCR4 regulates migration and development of human acute myelogenous leukemia stem cells in transplanted NOD/SCID mice. Cancer Res. 64, 2817–2824 (2004)

    Article  CAS  PubMed  Google Scholar 

  13. Spiegel, A. et al. Unique SDF-1-induced activation of human precursor-B ALL cells as a result of altered CXCR4 expression and signaling. Blood 103, 2900–2907 (2004)

    Article  CAS  PubMed  Google Scholar 

  14. Darash-Yahana, M. et al. Role of high expression levels of CXCR4 in tumor growth, vascularization, and metastasis. FASEB J. 18, 1240–1242 (2004)

    Article  CAS  PubMed  Google Scholar 

  15. Spano, J. P. et al. Chemokine receptor CXCR4 and early-stage non-small cell lung cancer: pattern of expression and correlation with outcome. Ann. Oncol. 15, 613–617 (2004)

    Article  PubMed  Google Scholar 

  16. Ma, Q. et al. Impaired B-lymphopoiesis, myelopoiesis, and derailed cerebellar neuron migration in CXCR4- and SDF-1-deficient mice. Proc. Natl Acad. Sci. USA 95, 9448–9453 (1998)

    Article  ADS  CAS  PubMed  PubMed Central  Google Scholar 

  17. Peled, A. et al. Dependence of human stem cell engraftment and repopulation of NOD/SCID mice on CXCR4. Science 283, 845–848 (1999)

    Article  ADS  CAS  PubMed  Google Scholar 

  18. Mazo, I. B. & von Andrian, U. H. Adhesion and homing of blood-borne cells in bone marrow microvessels. J. Leukoc. Biol. 66, 25–32 (1999)

    Article  CAS  PubMed  Google Scholar 

  19. Hatse, S., Princen, K., Bridger, G., De Clercq, E. & Schols, D. Chemokine receptor inhibition by AMD3100 is strictly confined to CXCR4. FEBS Lett. 527, 255–262 (2002)

    Article  CAS  PubMed  Google Scholar 

  20. Liles, W. C. et al. Mobilization of hematopoietic progenitor cells in healthy volunteers by AMD3100, a CXCR4 antagonist. Blood 102, 2728–2730 (2003)

    Article  CAS  PubMed  Google Scholar 

  21. Peled, A. et al. The chemokine SDF-1 stimulates integrin-mediated arrest of CD34+ cells on vascular endothelium under shear flow. J. Clin. Invest. 104, 1199–1211 (1999)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Jo, D. Y., Hwang, J. H., Kim, J. M., Yun, H. J. & Kim, S. Human bone marrow endothelial cells elaborate non-stromal-cell-derived factor-1 (SDF-1)-dependent chemoattraction and SDF-1-dependent transmigration of hematopoietic progenitors. Br. J. Haematol. 121, 649–652 (2003)

    Article  CAS  PubMed  Google Scholar 

  23. Novak, J., Georgakoudi, I., Wei, X., Prossin, A. & Lin, C. P. In vivo flow cytometer for real-time detection and quantification of circulating cells. Opt. Lett. 29, 77–79 (2004)

    Article  ADS  CAS  PubMed  PubMed Central  Google Scholar 

  24. Hendrix, C. W. et al. Pharmacokinetics and safety of AMD-3100, a novel antagonist of the CXCR-4 chemokine receptor, in human volunteers. Antimicrob. Agents Chemother. 44, 1667–1673 (2000)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Bleul, C. C., Fuhlbrigge, R. C., Casasnovas, J. M., Aiuti, A. & Springer, T. A. A highly efficacious lymphocyte chemoattractant, stromal cell-derived factor 1 (SDF-1). J. Exp. Med. 184, 1101–1109 (1996)

    Article  CAS  PubMed  Google Scholar 

  26. Bleul, C. C., Wu, L., Hoxie, J. A., Springer, T. A. & Mackay, C. R. The HIV coreceptors CXCR4 and CCR5 are differentially expressed and regulated on human T lymphocytes. Proc. Natl Acad. Sci. USA 94, 1925–1930 (1997)

    Article  ADS  CAS  PubMed  PubMed Central  Google Scholar 

  27. Christopherson, K. W., Hangoc, G., Mantel, C. R. & Broxmeyer, H. E. Modulation of hematopoietic stem cell homing and engraftment by CD26. Science 305, 1000–1003 (2004)

    Article  ADS  CAS  PubMed  Google Scholar 

  28. Mastro, A. M., Gay, C. V. & Welch, D. R. The skeleton as a unique environment for breast cancer cells. Clin. Exp. Metastasis 20, 275–284 (2003)

    Article  CAS  PubMed  Google Scholar 

  29. Cao, Y. A. et al. Shifting foci of hematopoiesis during reconstitution from single stem cells. Proc. Natl Acad. Sci. USA 101, 221–226 (2004)

    Article  ADS  CAS  PubMed  Google Scholar 

  30. Shen, H. et al. CXCR-4 desensitization is associated with tissue localization of hemopoietic progenitor cells. J. Immunol. 166, 5027–5033 (2001)

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank I. Alley, G. Adams, R. Klein and D. Worhunsky for help with stem cell harvesting and purification, and C. Pitsillides for assistance with T-cell purification techniques. We are grateful to D. Dombkowski for assistance with cell sorting. Special thanks to S. Harvey for assistance with illustrations and to A. Chenn for review of the manuscript. This work was supported by National Institute of Health (NIH) grants to C.P.L. and D.T.S., a NIH training grant (D.A.S.) and a Whitaker Foundation Graduate Fellowship (J.W.W.).

Author information

Authors and Affiliations

  1. Wellman Center for Photomedicine,

    Dorothy A. Sipkins, Xunbin Wei, Juwell W. Wu, Judith M. Runnels, Daniel Côté & Charles P. Lin

  2. Center for Regenerative Medicine and Technology, Massachusetts General Hospital, Harvard Medical School, 55 Fruit St, 02114, Boston, Massachusetts, USA

    Dorothy A. Sipkins & David T. Scadden

  3. Department of Hematology-Oncology, Massachusetts General Hospital and Dana-Farber Cancer Institute, Harvard Medical School,

    Dorothy A. Sipkins & David T. Scadden

  4. Center for Immunology and Inflammatory Diseases, Massachusetts General Hospital East, Harvard Medical School, Building 149, 13th Street, Massachusetts, 02129, Charlestown, USA

    Terry K. Means & Andrew D. Luster

  5. Harvard Stem Cell Institute, Harvard University, Massachusetts, 02138, Cambridge, USA

    David T. Scadden

Authors
  1. Dorothy A. Sipkins
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Xunbin Wei
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Juwell W. Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Judith M. Runnels
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Daniel Côté
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Terry K. Means
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Andrew D. Luster
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. David T. Scadden
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Charles P. Lin
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Dorothy A. Sipkins or Charles P. Lin.

Ethics declarations

Competing interests

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

Supplementary information

Supplementary Video S1

This video shows Nalm-6 leukemic cells rolling along but not adhering to skin vasculature. (MOV 2070 kb)

Supplementary Video S2

This movie shows Nalm-6 leukemic cells adhering to specific vascular domains in the bone marrow. (MOV 9159 kb)

Supplementary Video S3

This movie shows another view of Nalm-6 leukemic homing interactions in a bone marrow parasagittal vascular bed. (MOV 9281 kb)

Supplementary Video S4

This movie shows footage of extensive Nalm-6 rolling and binding interactions in a large frontal vein possessing high blood flow-rate. (MOV 10144 kb)

Supplementary Figure S1

This figure shows that MatLyLu (MLL) prostatic carcinoma cells home to the same vascular microdomains as Nalm-6 leukemic cells. (JPG 28 kb)

Supplementary Figure S2

This figure shows that Nalm-6 binding and SDF-1 expression occur in restricted regions of bone marrow. SDF-1+ regions are contrasted with SDF-1- areas. Corresponding Nalm-6 homing patterns are shown. (JPG 31 kb)

Supplementary Figure S3

This figure shows the restricted pattern of SDF-1 vascular expression in bone marrow by in vivo double-labeling of the vasculature with fluorescent SDF-1 and PECAM-1. (JPG 62 kb)

Supplementary Figure S4

This figure shows that Nalm-6 homing is inhibited after SDF-1-desensitization treatment. (JPG 18 kb)

Supplementary Figure S5

This figure shows that CXCR4 blockade causes a modest mobilization of engrafted Nalm-6 leukemic cells into the peripheral circulation. (JPG 19 kb)

Supplementary Figure S6

This figure shows in vitro studies that confirm the relative persistence of fluorescence signal in slowly dividing cells, as detected by flow cytometry. (JPG 23 kb)

Supplementary Figure S7

This figure shows that benign T lymphocytes home to the same SDF-1+ vascular microdomains as Nalm-6 leukemic cells. (JPG 17 kb)

Supplementary Figure S8

This figure shows that VLA-4/VCAM-1 interactions do not appear critical for initial homing of Nalm-6 pre-B ALL to bone marrow. (JPG 24 kb)

Supplementary Figure S9

This figure shows that Nalm-6 leukemic cells pre-treated with N-Cadherin blocking antibody show altered homing kinetics compared to control. (JPG 31 kb)

Supplementary Figure S10

This figure demonstrates that differences in signal intensity detection by in vivo immunoimaging appear to result from variations in optical properties of the surrounding tissue. (JPG 46 kb)

Supplementary Figures and Supplementary Videos Legends

Legends to accompany the above Supplementary Figures and Supplementary Videos (DOC 32 kb)

Supplementary Methods (DOC 35 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Sipkins, D., Wei, X., Wu, J. et al. In vivo imaging of specialized bone marrow endothelial microdomains for tumour engraftment. Nature 435, 969–973 (2005). https://doi.org/10.1038/nature03703

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nature03703

This article is cited by

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing