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Signaling for lymphangiogenesis via VEGFR-3 is required for the early events of metastasis

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Abstract

Metastasis to regional lymph nodes is an important and early event in many tumors. Vascular endothelial growth factor-C (VEGF-C), VEGF-D and their receptor VEGFR-3, play a role in tumor spread via the lymphatics, although the timing of their involvement is not understood. In contrast, VEGFR-2, activated by VEGF-A, VEGF-C and VEGF-D, is a mediator of angiogenesis and drives primary tumor growth. We demonstrate the critical role for VEGFR-3, but not VEGFR-2, in the early events of metastasis. In a tumor model exhibiting both VEGF-D-dependent angiogenesis and lymphangiogenesis, an antibody to VEGFR-2 (DC101) was capable of inhibiting angiogenesis (79 % reduction in PECAM + blood vessels) and growth (93 % reduction in tumor volume). However, unlike an anti-VEGFR-3 Mab (mF4-31C1), DC101 was not capable of eliminating either tumor lymphangiogenesis or lymphogenous metastasis (60 % reduction of lymph node metastasis by DC101 vs 95 % by mF4-31C1). Early excision of the primary tumors demonstrated that VEGF-D-mediated tumor spread precedes angiogenesis-induced growth. Small but highly metastatic primary human breast cancers had significantly higher lymphatic vessel density (23.1 vessels/mm2) than size-matched (11.7) or larger non-metastatic tumors (12.4) thus supporting the importance of lymphatic vessels, as opposed to angiogenesis-mediated primary tumor growth, for nodal metastasis. These results suggest that lymphangiogenesis via VEGF-D is more critical than angiogenesis for nodal metastasis.

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Abbreviations

LN:

Lymph node

LVD:

Lymphatic vessel density

LYVE-1:

Lymphatic vessel endothelial receptor 1

NK:

Natural killer

Mab:

Monoclonal antibody

MTT:

3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide

PBS:

Phosphate-buffered saline

PECAM-1:

Platelet endothelial cell adhesion molecule-1

SCID/NOD:

Severe combined immunodeficiency/non-obese diabetic

VEGF:

Vascular endothelial growth factor

VEGFR:

Vascular endothelial growth factor receptor

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Acknowledgments

We thank You-Fang Zhang for scientific and technical help with immunohistochemistry, and the staff of the Animal Facility at the Ludwig Institute for Cancer Research for assistance with SCID/NOD mice. We also thank Anthony Burgess for helpful discussion and critical reading of this manuscript.

Funding

This work was supported by Program Grants and Research Fellowships from the National Health and Medical Research Council of Australia and by funds from the Operational Infrastructure Support Program, Victorian Government, Australia.

Conflict of interest

SAS and MGA are on the Scientific Advisory Board of Vegenics Ltd, a wholly owned subsidiary of Circadian Technologies, are stock holders and consultants.

Author information

Author notes

  1. Masataka Matsumoto

    Present address: Department of Surgical Oncology and Digestive Surgery, Field of Oncology Course of Advanced Therapeutics, Graduate School of Medical and Dental Sciences, Kagoshima University, 8-35-1, Sakuragaoka, Kagoshima, 890-8520, Japan

  2. Teruhiko Sato

    Present address: Department of Respiratory Medicine, School of Medicine, Juntendo University, 2-1-1 Hongo, Bunkyo-Ku, Tokyo, 113-8421, Japan

Authors and Affiliations

  1. Ludwig Institute for Cancer Research, Royal Melbourne Hospital, P.O. Box 2008, Parkville, VIC, 3050, Australia

    Masataka Matsumoto, Sally Roufail, Rachael Inder, Carol Caesar, Tara Karnezis, Ramin Shayan, Rae H. Farnsworth, Teruhiko Sato, Marc G. Achen & Steven A. Stacker

  2. Department of Surgery, Royal Melbourne Hospital, University of Melbourne, Parkville, VIC, 3050, Australia

    Ramin Shayan, Rae H. Farnsworth & G. Bruce Mann

  3. Royal Womens Hospital, Parkville, VIC, 3052, Australia

    G. Bruce Mann

  4. Tumour Angiogenesis Program, Peter MacCallum Cancer Centre, Locked bag 1, A’Beckett Street, East Melbourne, VIC, 8006, Australia

    Sally Roufail, Carol Caesar, Tara Karnezis, Ramin Shayan, Rae H. Farnsworth, Marc G. Achen & Steven A. Stacker

  5. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, VIC, 3010, Australia

    Marc G. Achen & Steven A. Stacker

Authors
  1. Masataka Matsumoto
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  2. Sally Roufail
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Corresponding author

Correspondence to Steven A. Stacker.

Additional information

Steven A. Stacker and Marc G. Achen are on the Scientific Advisory Board of Vegenics Ltd, a wholly owned subsidiary of Circadian Technologies, are stock holders and consultants.

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Matsumoto, M., Roufail, S., Inder, R. et al. Signaling for lymphangiogenesis via VEGFR-3 is required for the early events of metastasis. Clin Exp Metastasis 30, 819–832 (2013). https://doi.org/10.1007/s10585-013-9581-x

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  • DOI: https://doi.org/10.1007/s10585-013-9581-x

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