Abstract 3484: Murine and human lymphatic endothelial cells show similar marker profiles and migratory properties

Abstract

Abstract Introduction: Lymphangiogenesis (growth of nascent lymphatic vessels in vivo and proliferation, migration and tube formation of lymphatic endothelial cells (LECs) in vitro) plays a key role in lymph node metastasis in a number of tumour types (1), including prostate cancer. Mechanisms underlying tumor cell-lymphatic vessel interactions are critical to understand. Thus determining the molecular basis of LEC motility and its relationship to lymphangiogenesis and subsequent metastasis will play an important part in developing novel strategies to prevent metastasis. We have previously shown that members of the vascular endothelial growth factor (VEGF) family regulate migration of human LECs (2). As the effect of stimulation with VEGF family ligands has not been reported in murine LECs, this study aims to isolate and characterize murine LECs and to compare their migratory profiles to human LECs. This data will be important in determining the relevance of experiments utilizing LECs derived from transgenic and/or human tumor-bearing mice to the study of lymphangiogenesis in human cancers. Results: Murine LECs were successfully isolated from prostate and skin using a CD34 depletion, CD31 positive selection strategy. Murine LECs expressed lymphatic endothelial cell markers Prox-1 and LYVE-1. The migratory profile of murine LECs was compared to human LECs using a panel of extracellular substrates and VEGF family ligands. VEGF-C and VEGF-D potently stimulated murine LEC migration in vitro. Stimulation of murine LECs with VEGF-C caused activation of pAKT(Ser347), Erk1/2 and JNK signalling pathways, consistent with observations in human LECs. Conclusions: This study describes a successful, robust protocol for isolation of murine LECs. Murine LECs derived from transgenic and knock-out mouse models will provide important insights into the molecular mechanisms of lymphangiogenesis and allow the study of LECs in the context of cancer metastasis and other disease associated with altered lymphatic vessel architecture or function. (1) Mumprecht V, Detmar M (2009) J Cell Mol Med, 13: 1405-16 (2) Zeng Y, Opeskin K, Goad J, Williams ED (2006) Cancer Res, 66: 9566-75 Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 3484. doi:10.1158/1538-7445.AM2011-3484