A Multiscale Analysis of Early Cardiogenesis Following VEGF Perturbations

• DOI: 10.1096/fasebj.2018.32.1_supplement.94.1
• Publication Date: 2021-06-21T16:38:04Z
• Authors: Brenda J. Rongish; Anastasiia Aleksa...; Michael B. Filla; Edina Kosa; Charles D. Little; Alan Petersen; Andras Czirok; Sandra Rugonyi; Graham Rykiel; Rusty Lansford
• Keywords: 0301 basic medicine; 03 medical and health sciences; 3. Good health

Congenital Heart Defects (CHDs) are multi‐factorial in origin, and there is a critical need to identify other contributing factors in the development of CHDs. Endogenous factors that regulate development, such as altered levels of cardiovascular growth factors in the embryo/fetus, may initiate or contribute to CHDs. Our objective was to determine if alteration of Vascular Endothelial Growth Factor (VEGF) levels at key developmental periods leads to reproducible cardiovascular defects, which may be correlated with underlying altered cell behavior, physiological parameters and/or molecular signaling events. We used ex ovo culture and imaging systems that capitalize on the experimental accessibility of quail embryos, which, like humans, have four‐chambered hearts. In order to alter VEGF signaling, the following were introduced into quail embryos: recombinant human VEGF, exogenous VEGF‐GFP fusion protein, and inhibitors of VEGF receptor tyrosine kinase signaling and downstream effectors of VEGF. Time‐lapse imaging (KUMC) and optical coherence tomography (OHSU) revealed abnormal phenotypes between Hamburger and Hamilton (1951) stages 4 to HH38 – encompassing vasculogenic (pre‐circulatory) stages through primordial valve formation. Hemodynamic parameters were measured and related to structural defects at circulation stages. These studies showed the effects of altered VEGF signaling on cardiac and endothelial cellular behavior, organogenesis of four‐chambered hearts and great vessels, and cardiac function. Further, we identified potential molecular pathways involved in VEGF signaling. The ability to observe the behavior of endothelial, endocardial, and myocardial precursors, gross cardiac phenotypes and hemodynamic changes following experimental manipulation of VEGF allowed a careful analysis of the role of VEGF at multiple length scales, and over time, during cardiovascular morphogenesis. We conclude that VEGF signaling, when dysregulated, leads to detrimental cardiovascular malformations and altered cardiac cellular behavior and function. This work has helped to determine the underlying mechanisms of VEGF's role and actions in cardiac and vessel development, as well as identified structural and functional cardiovascular defects that are attributable to aberrant VEGF signaling; which are similar to CHDs observed in humans.Support or Funding InformationAmerican Heart Association 16GRNT29840002, the Molecular Regulation of Cell Development and Differentiation COBRE, P30 GM122731, and the University of Kansas School of Medicine Bridging Award.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.