Complementary Wnt Sources Regulate Lymphatic Vascular Development via PROX1-Dependent Wnt/β-Catenin Signaling

Summary: Wnt/β-catenin signaling is necessary for lymphatic vascular development. Oscillatory shear stress (OSS) enhances Wnt/β-catenin signaling in cultured lymphatic endothelial cells (LECs) to induce expression of the lymphedema-associated transcription factors GATA2 and FOXC2. However, the mechanisms by which OSS regulates Wnt/β-catenin signaling and GATA2 and FOXC2 expression are unknown. We show that OSS activates autocrine Wnt/β-catenin signaling in LECs in vitro. Tissue-specific deletion of Wntless, which is required for the secretion of Wnt ligands, reveals that LECs and vascular smooth muscle cells are complementary sources of Wnt ligands that regulate lymphatic vascular development in vivo. Further, the LEC master transcription factor PROX1 forms a complex with β-catenin and the TCF/LEF transcription factor TCF7L1 to enhance Wnt/β-catenin signaling and promote FOXC2 and GATA2 expression in LECs. Thus, our work defines Wnt sources, reveals that PROX1 directs cell fate by acting as a Wnt signaling component, and dissects the mechanisms of PROX1 and Wnt synergy. : Cha et al. demonstrate that lymphatic vascular development is regulated by Wnt ligands secreted by lymphatic endothelial cells (LECs) and vascular smooth muscle cells. Oscillatory shear stress regulates Wnt secretion from LECs. PROX1 interacts with β-catenin and TCF7L1 to regulate Wnt signaling and promote the expression of FOXC2 and GATA2. Keywords: lymphatic vasculature, lymphovenous valves, lymphatic valves, PROX1, GATA2, FOXC2, Wnt/β-catenin signaling, oscillatory shear stress, Wntless, TCF7L