The study of neuro-vascular interactions in the developing heart

National Heart, Lung, And Blood Institute

Investigators

Linked publications & trials

Abstract

1) Sympathetic control of cardiomyocyte differentiation and maturation. To determine whether sympathetic innervation influences cardiomyocyte differentiation and maturation, we have developed a 3D co-culture system of GFP- and GCaMP6-expressing iPS-derived cardiomyocytes with sympathetic neurons, endothelial cells, and epicardial cells. Our findings show that co-cultured cardiomyocytes exhibit enhanced functional, structural, and gene expression properties (Kowalski et al. PMID: 35359438). Currently, we are working on establishing photo-activated regulation of intracellular cAMP in sympathetic neurons to manipulate the adrenergic signaling pathway. This will allow us to investigate whether sympathetic activation affects cardiomyocyte maturation. These studies aim to provide a mechanistic understanding of the role of sympathetic innervation in the functional development of the embryonic heart. 2) Heart-dependent blood circulation is essential for vascular development. We are investigating the factors that drive the transition from a capillary plexus to a hierarchical vascular network during vascular development. To explore this process, we have developed a protocol for generating vascular organoids from mouse embryonic stem cells (ESCs) and transplanting them onto the chick chorioallantoic membrane (CAM). In vitro, these organoids form a three-dimensional capillary plexus, comprising endothelial cells and pericytes, which closely resembles the vasculature found in E13.5 skin and coronary tissues. After transplantation onto the day 7 chick CAM, the vascular organoids undergo angiogenesis and remodeling. We observe the formation of branched hierarchical networks containing large-diameter, smooth muscle-covered blood vessels derived from the organoid ESCs, resembling the E15.5 skin and coronary vasculature. Overall, our vascular organoid model effectively recapitulates embryonic vasculogenesis and capillary plexus formation in vitro, followed by subsequent angiogenesis and remodeling after transplantation to the chick CAM in vivo (Kowalski et al. PMID: 40021912). Additionally, we plan to use our vascular organoids to generate assembloids that include cardiomyocytes and sympathetic neurons.