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

Stimulation of lymphangiogenesis via VEGFR-3 inhibits chronic skin inflammation

“…Importantly, both of the latter models are strongly TNF dependent. Conversely, application of VEGF-C or tumor-derived VEGF-C have been found to enhance lymphatic function and flow, respectively Szuba et al, 2002;Yoon et al, 2003;Hoshida et al, 2006;Tammela et al, 2007). The impaired lymphatic drainage might also explain the accumulation of CD11b + cells that we observed after blockade of VEGFR-3.…”

The New Era of the Lymphatic System: No Longer Secondary to the Blood Vascular System

“…Administration of a VEGF-C-expressing adenovirus enhanced generation of functional lymphatic vessels in Chy mutant mice (Karkkainen et al 2001), and adenoviral delivery of VEGF-C in normal skin or at the edge of epigastric skin flaps in mice strongly induced lymphangiogenesis (Enholm et al 2001;Saaristo et al 2004). In addition, recombinant VEGF-C was shown to be sufficient to activate in vivo lymphangiogenesis and reversed the symptoms of lymphedema in rabbit ears (Szuba et al 2002); VEGF-C gene therapy stimulated postnatal lymphangiogenesis and thus ameliorated secondary lymphedema in animal models (Yoon et al 2003). More recently, using a newly established mouse model in which the axillary lymph nodes were removed to better recapitulate human breast cancer lymphadenectomy, Tammela et al (2007) have shown that collecting lymphatic vessels could be efficiently regenerated and connected to transplanted lymph node in the lymphedema area.…”

Tratamento com VEGFC para revascularização linfática em membros pélvicos de camundongos

“…As plaquetas se agregam em locais de comunicação entre a veia cardinal e sacos linfáticos e "sela" os vasos linfáticos da veia cardinal, separando os dois vasos (SCHULTE-MERKER; SABINE; PETROVA, 2011). As plaquetas são importantes para manter esses dois sistemas vasculares separados: a depleção de plaquetas ou defeito de agregação plaquetária leva à conexões linfovenosas anormais e à vasos linfáticos cheios de sangue (ICHISE et al, 2009;BERTOZZI et al, 2010;CARRAMOLINO et al, 2010;SUZUKI-INOUE et al, 2010;UHRIN et al, 2010 ROCKSON, 1998;YOON et al, 2003;CHEUNG et al, 2006;SAITO et al, 2006 ROCKSON, 1998;KARKKAINEN et al, 2001;YOON et al, 2003;SAARISTO et al, 2004;CHEUNG et al, 2006;SAARISTO et al, 2006;TAMMELA et al, 2007) (APRELIKOVA et al, 1992;GALLAND et al, 1992;JOUKOV et al, 1996;LEE et al, 1996). Apesar do VEGFR3 desempenhar um papel fundamental para o desenvolvimento celular endotelial vascular e linfático, sua expressão fica limitada ao endotélio linfático nos estágios finais de desenvolvimento (KAIPAINEN et al, 1995;KUKK et al, 1996;BAUMGARTNER et al, 1998 FERRELL et al, 1998;KARKKAINEN et al, 2000).…”

“…Porém, com a realização do tratamento com VEGFC, pode-se observar um aumento significativo em ambas essas variáveis para a pele, indicando que o tratamento com VEGFC causou mudanças significativas na revascularização linfática nesta região (CHEUNG et al, 2006;PAN et al, 2006 (OASHI et al, 2012;KUWAHARA et al, 2013;OASHI et al, 2013). Outra possibilidade pode ser o fato da proximidade dessa região com o local de aplicação do VEGFC exógeno, uma vez que a aplicação foi realizada intraperitonealmente próxima à região da cicatriz cirúrgica (YOON et al, 2003).…”

“…Recentemente, tem sido relatado que a super-expressão de VEGFC (JIN et al, 2009;OASHI et al, 2012) A maioria dos relatos anteriores de terapia de linfedemas com fator de crescimento é baseada em modelos de camundongo, os quais têm várias limitações quando se considera a sua tradução em clínica para o tratamento de pacientes humanos (RUTKOWSKI et al, 2006;LALONI, 2008;WADA et al, 2011). Primeiramente, as condições hidrostáticas são dramaticamente diferentes em camundongos porque os seres humanos são consideravelmente mais altos SABINE;PETROVA, 2011;OASHI et al, 2012;MARCHIÒ;ASTANINA;BUSSOLINO, 2013 (YOON et al, 2003;SWARTZ, 2006;RUTKOWSKI et al, 2006;LALONI, 2008;JIN et al, 2009;OASHI et al, 2012).…”