Vascular specification from diversified mesoderm culminates in endothelial convergence

Abstract

Vasculogenesis establishes the embryonic circulatory system, yet how a common vascular program emerges across multiple embryonic and extraembryonic developmental environments remains incompletely understood. Here, we reconstruct mouse vasculogenesis through integrated single-cell, spatial, lineage and perturbation analyses. We find that vascular specification emerges from distinct sources of pre-specified mesoderm through a transient Kdr+Etv2- vascular-primed state promoted by BMP signaling. Primed cells persist until E8.0- E8.5, after which they resolve into committed derivatives. Dual blood-endothelial potential is largely restricted to early vitelline progenitors, whereas embryonic and umbilical progenitors contribute predominantly to endothelial lineages. Temporal VEGF signaling perturbations validate the inferred specification sequence, placing VEGF downstream of vascular priming and demonstrating its indispensable role in vascular and primitive blood specification. Unlike most developmental lineages, endothelial derivatives converge toward a common transcriptional state while retaining transcriptional traces of their developmental origins. Together, these findings establish a mechanistically validated developmental sequence for mammalian vasculogenesis.