Genetic Alteration of Endothelial Heparan Sulfate Selectively Inhibits Tumor Angiogenesis

To examine the role of endothelial heparan sulfate during angiogenesis, we generated mice bearing an endothelial-tarqceted deletion in the biosyn- thetic enzyme N-acetylglucosamine N-deacetylase/ N-sulfotransferase 1 (Ndstl). Physiological angiogenesis during cutaneous wound repair was unaffected, as was growth and reproductive capacity of the mice. In contrast, pathological angiogenesis in experimental tumors was altered, resulting in smaller tumors and reduced microvascular density and branching. To simulate the angiogenic environment of the tumor, endothelial cells were isolated and propagated in vitro with proangiogenic growth factors. Binding of FGF-2 and$VEGF_{164}$to cells and to purified heparan sulfate was dramatically reduced. Mutant endothelial cells also exhibited altered sprouting responses to FGF-2 and$VEGF_{164}$, reduced Erk phosphorylation, and an increase in apoptosis in branching assays. Corresponding changes in growth factor binding to tumor endothelium and apoptosis were also observed in vivo. These findings demonstrate a cell-autonomous effect of heparan sulfate on endothelial cell growth in the context of tumor angiogenesis.