Spatiotemporal Atlas of Heart Development Reveals Blood-Flow-Dependent Cellular, Structural, Metabolic, and Spatial Remodeling

Abstract

Embryonic heart development depends on coordinated interactions between cellular programs, molecular signaling, and biomechanical forces, yet how mechanical cues shape cellular and molecular pathways remains incompletely understood. We perturbed cardiac blood flow by partial left or right atrial ligation (LAL/RAL) in chick embryos, generating chamber-specific hemodynamic gain- or loss-of-function states. Using single-cell and unbiased, high-resolution spatial transcriptomics, we generated a spatiotemporal atlas of flow-dependent tissue development, enabling systematic investigations of bidirectional interactions between blood-flow mechanics and tissue development. Spatially resolved analyses recapitulated key features of normal morphogenesis including regional maturation and the cellular neighborhood. We further revealed flow-specific remodeling across molecular, cellular, and architectural levels. Altered flow induced LOX-expressing cardiomyocyte and endocardial states, disrupted ventricular layer organization, and delayed maturation, alongside transient metabolic and ion-transport adaptations. Together, these findings define how redistributed blood flow reshapes developing cardiac tissues and provide a framework for studying flow-dependent remodeling in morphogenesis and malformation.