Spatial-temporal order-disorder transition in angiogenic NOTCH signaling controls cell fate specification

Angiogenesis is a morphogenic process resulting in the formation of new blood vessels from pre-existing ones, usually in hypoxic micro-environments. The initial steps of angiogenesis depend on robust differentiation of oligopotent endothelial cells into the Tip and Stalk phenotypic cell fates, controlled by NOTCH-dependent cell-cell communication. The dynamics of spatial patterning of this cell fate specification are only partially understood. Here, by combining a controlled experimental angiogenesis model with mathematical and computational analyses, we find that the regular spatial Tip-Stalk cell patterning can undergo an order-disorder transition at a relatively high input level of a pro-angiogenic factor VEGF. The resulting differentiation is robust but temporally unstable for most cells, with only a subset of presumptive Tip cells leading sprout extensions. We further find that sprouts form in a manner maximizing their mutual distance, consistent with a Turing-like model that may depend on local enrichment and depletion of fibronectin. Together, our data suggest that NOTCH signaling mediates a robust way of cell differentiation enabling but not instructing subsequent steps in angiogenic morphogenesis, which may require additional cues and self-organization mechanisms. This analysis can assist in further understating of cell plasticity underlying angiogenesis and other complex morphogenic processes.Significance StatementWe investigate the spatial and temporal patterns of Tip/Stalk specification and the ensuing angiogenic sprouting by using a novel controlled micro-engineered experimental model of angiogenesis and a set of mathematical models of the spatially resolved, cell population-level VEGF-NOTCH signaling. Our analysis provides a dynamic view of the initial step of angiogenesis, revealing fluctuations in its onset, and features suggesting transitions between order and disorder in cell organization. These findings suggest how a potentially very restrictive patterning mechanism can become sensitive to a variety of environmental cues. This sensitivity can be crucial for proper vascularization of a damaged organ, and may suggest new ways of analyzing angiogenesis in the context of cancer and other pathologies. This analysis also suggests a framework for understanding of other instances of NOTCH-mediated patterning processes.