Mapping the Role of the Notch4 Receptor in Angiogenesis

Angiogenesis is the process by which new blood vessels form, as cells in hypoxia secret Vascular Endothelial Growth Factor which creates the tip cell phenotype in whichever endothelial cell it binds to. The new tip cell promotes the stalk cell phenotype in neighboring cells through the Notch pathway. The tip and stalk cell phenotypes are critical to angiogenesis, but not much is understood about the pathway responsible for their creation. With this experiment, we aim to understand how the pathway functions by examining the impact of knocking out components of it, namely Notch4 and its downstream targets, Rnd1 and Unc5b. In order to quantify the effects of knocking out these proteins using CRISPR/Cas9, postnatal mice retina were compared on measures of radial outgrowth, tip cells, and branching of the retinal vasculature. Based on initial results, it is expected that the Notch4 knockout mice will exhibit less radial outgrowth and reduced branching of retinal blood vessels. The mapping of angiogenic pathways offers many avenues for studying human health. For example, new cancer research aims to reduce blood flow to tumors by decreasing angiogenesis. Therefore, a deeper understanding of the role of Notch4 could aid the development of new medicinal therapies