VEGF-C gene therapy augments postnatal lymphangiogenesis and ameliorates secondary lymphedema
Young-sup Yoon, … , Jeffrey M. Isner, Douglas W. Losordo
Young-sup Yoon, … , Jeffrey M. Isner, Douglas W. Losordo
Published March 1, 2003
Citation Information: J Clin Invest. 2003;111(5):717-725. https://doi.org/10.1172/JCI15830.
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VEGF-C gene therapy augments postnatal lymphangiogenesis and ameliorates secondary lymphedema

Abstract

Although lymphedema is a common clinical condition, treatment for this disabling condition remains limited and largely ineffective. Recently, it has been reported that overexpression of VEGF-C correlates with increased lymphatic vessel growth (lymphangiogenesis). However, the effect of VEGF-C–induced lymphangiogenesis on lymphedema has yet to be demonstrated. Here we investigated the impact of local transfer of naked plasmid DNA encoding human VEGF-C (phVEGF-C) on two animal models of lymphedema: one in the rabbit ear and the other in the mouse tail. In a rabbit model, following local phVEGF-C gene transfer, VEGFR-3 expression was significantly increased. This gene transfer led to a decrease in thickness and volume of lymphedema, improvement of lymphatic function demonstrated by serial lymphoscintigraphy, and finally, attenuation of the fibrofatty changes of the skin, the final consequences of lymphedema. The favorable effect of phVEGF-C on lymphedema was reconfirmed in a mouse tail model. Immunohistochemical analysis using lymphatic-specific markers: VEGFR-3, lymphatic endothelial hyaluronan receptor-1, together with the proliferation marker Ki-67 Ab revealed that phVEGF-C transfection potently induced new lymphatic vessel growth. This study, we believe for the first time, documents that gene transfer of phVEGF-C resolves lymphedema through direct augmentation of lymphangiogenesis. This novel therapeutic strategy may merit clinical investigation in patients with lymphedema.

Authors

Young-sup Yoon, Toshinori Murayama, Edwin Gravereaux, Tengiz Tkebuchava, Marcy Silver, Cynthia Curry, Andrea Wecker, Rudolf Kirchmair, Chun Song Hu, Marianne Kearney, Alan Ashare, David G. Jackson, Hajime Kubo, Jeffrey M. Isner, Douglas W. Losordo

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Figure 5

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(a–j) phVEGF-C induces proliferation of lymphatic endothelial cells. Dou...
(a–j) phVEGF-C induces proliferation of lymphatic endothelial cells. Double immunohistochemistry using LYVE-1 and Ki-67 in active lymphangiogenesis site from skin sections. In a, d, and g, LYVE-1 staining of lymphatic vessels (arrows) in the dermis. In b, e, and h, green fluorescence (white arrowheads) depicts the nuclear staining of Ki-67. In c, f, and i, double fluorescence (yellow arrowheads) demonstrates Ki-67+ nuclei (green) in lymphatic vessels (red). Lymphatic vessels in normal skin (c) are shown negative for Ki-67. In the LacZ group, some of the lymphatic vessels contain Ki-67+ nuclei (f). White arrows in f show Ki-67– lymphatic vessels. In phVEGF-C–transfected skin, most of the LYVE-1–positive lymphatic vessels are positive for Ki-67 (i), indicating that active cell division occurs in the lymphatic vessels. (j) Number of Ki-67+ nuclei are 2.5 times higher in the VEGF-C group. *P < 0.01 compared with normal; **P < 0.01 compared with saline and LacZ. Scale bar, 100 μm. (k–u) phVEGF-C does not increase capillary density in two animal models of lymphedema. Immunohistochemistry with CD31 (PECAM-1) in a rabbit ear (k–n) and a mouse tail (p–t) model of lymphedema on skin sections from the normal (k and p), saline (l and q), LacZ (m and r), VEGF-C (n and s), and VEGF165 (t) groups. Vascular endothelial cells are stained red (black arrows). o and u show quantification of capillary density. Only the VEGF165 group in the mouse tail model demonstrated significantly higher capillary density than the other groups. *P < 0.01 vs. saline, LacZ, and VEGF-C. Scale bar, 100 μm.