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Therapeutic differentiation and maturation of lymphatic vessels after lymph node dissection and transplantation

Nature Medicine volume 13, pages 1458–1466 (2007)Cite this article

Abstract

Surgery or radiation therapy of metastatic cancer often damages lymph nodes, leading to secondary lymphedema. Here we show, using a newly established mouse model, that collecting lymphatic vessels can be regenerated and fused to lymph node transplants after lymph node removal. Treatment of lymph node–excised mice with adenovirally delivered vascular endothelial growth factor-C (VEGF-C) or VEGF-D induced robust growth of the lymphatic capillaries, which gradually underwent intrinsic remodeling, differentiation and maturation into functional collecting lymphatic vessels, including the formation of uniform endothelial cell-cell junctions and intraluminal valves. The vessels also reacquired pericyte contacts, which downregulated lymphatic capillary markers during vessel maturation. Growth factor therapy improved the outcome of lymph node transplantation, including functional reconstitution of the immunological barrier against tumor metastasis. These results show that growth factor–induced maturation of lymphatic vessels is possible in adult mice and provide a basis for future therapy of lymphedema.

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Figure 1: Functional maturation of a newly formed lymphatic vessel network and amelioration of lymphedema after lymph node removal in AdVEGF-C/D–transduced mice.
Figure 2: Short-term VEGF-C stimulation induces endothelial sprouting and leakage in collecting lymphatic vessels.
Figure 3: Maturation of lymphatic endothelial junctions in VEGF-C treated axillas.
Figure 4: Newly generated lymphatic endothelium becomes associated with SMCs, which regulate expression of the lymphatic capillary marker LYVE-1.
Figure 5: Lymphatic valves form in VEGF-C–induced collecting lymphatic vessels.
Figure 6: AdVEGF-C–transduced lymph node transplants incorporate into the existing lymphatic vasculature and trap metastatic tumor cells.

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Acknowledgements

We thank M. Achen and S. Stacker for the VEGF-DΔNΔC construct; Y. Cao for the PDGF-B construct; P. Korpisalo for the PDGF-B adenovirus; D. Kerjaschki for human podoplanin antibodies; M. Takeichi and H. Semb for the N-cadherin antibody; C. Heckman and K. Helenius for comments on the manuscript; the Biomedicum Molecular Imaging Unit for microscope support; and M. Helanterä, P. Hyvärinen, S. Lampi, K. Makkonen, A. Malinen, T. Tainola, S. Wallin and W. Zheng for technical assistance. This work was supported by grants from the National Institutes of Health (5 R01 HL075183-02) and The European Union (Lymphangiogenomics, LSHG-CT-2004-503573). T.T. has been supported by the Biomedicum Helsinki Foundation, the Finnish Cancer Organizations, the Finnish Cultural foundation, Nylands Nation and the Paulo Foundation as well as the Helsinki Biomedical Graduate School.

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Authors and Affiliations

  1. Molecular/Cancer Biology Laboratory, Haartman Institute and Ludwig Institute for Cancer Research, Biomedicum Helsinki, PO Box 63 (Haartmaninkatu 8), 00014 University of Helsinki, Finland

    Tuomas Tammela, Anne Saaristo, Tanja Holopainen, Johannes Lyytikkä, Anna Kotronen & Kari Alitalo

  2. Department of Plastic Surgery, Turku University Central Hospital, PO Box 52 (Kiinanmyllynkatu 8), Turku, 20521, Finland

    Anne Saaristo

  3. Department of Neurology, Experimental MRI Laboratory, Helsinki University Central Hospital, PO Box 63 (Haartmaninkatu 8), Helsinki, Finland.,

    Miia Pitkonen & Usama Abo-Ramadan

  4. Department of Biotechnology and Molecular Medicine, University of Kuopio, A. I. Virtanen Institute for Molecular Sciences, PO Box 1627, 70211 Kuopio, Finland.,

    Seppo Ylä-Herttuala

  5. Development and Differentiation Laboratory, Biomedicum Helsinki, PO Box 63 (Haartmaninkatu 8), 00014 University of Helsinki, Finland

    Tatiana V Petrova

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Contributions

T.T. designed, directed and performed mouse ear, paw and tumor experiments, surgery, cell culture, immunohistochemistry and data analysis, as well as interpreted results, and wrote the paper; A.S. designed, directed and performed surgery, performed tumor experiments, analyzed data, interpreted results and wrote the paper; T.H. performed paw and tumor experiments, cell culture, immunohistochemistry and data analysis, and helped perform surgery; J.L. performed immunohistochemistry, data analysis and 3D image rendering; A.K. performed immunohistochemistry and data analysis; M.P. performed MRI imaging and data analysis; U.A.-R. designed, directed and performed MRI imaging and data analysis; S.Y.-H. developed and provided adenovirus vectors; T.V.P. generated and provided reagents and helped write the paper; K.A. designed experiments, interpreted results and wrote the paper.

Corresponding author

Correspondence to Kari Alitalo.

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Competing interests

K.A. and S.Y.-H. are minor shareholders and boardmembers of Lymphatix, Ltd.

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Supplementary Table 1, Supplementary Figs. 1–9 and Supplementary Methods (PDF 1454 kb)

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Tammela, T., Saaristo, A., Holopainen, T. et al. Therapeutic differentiation and maturation of lymphatic vessels after lymph node dissection and transplantation. Nat Med 13, 1458–1466 (2007). https://doi.org/10.1038/nm1689

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