During the period of the past award, we have developed a transgenic mouse that targets Cre-recombinase to the epicardium. This is a unique model system that we have used to demonstrate that conditional mutation of the retinoid receptor alpha (RXR1) in the epicardium affects cardiac morphogenesis and impairs embryonic survival. This mutation also uncovered a novel role of nuclear receptors in coronary arteriogenesis. We have subsequently determined that the 2-catenin gene acts downstream of the RXR1 pathway and we have generated data demonstrating that mutation of 2-catenin in the epicardium impairs cardiac morphogenesis by reducing cardiac cell proliferation and coronary vessel formation. Here we hypothesize that a Wnt/2 catenin pathway in the epicardium regulates key steps in cardiac development, including cell proliferation in the myocardium and the formation of the coronary vasculature. To demonstrate this hypothesis, we propose three independent genetic approaches that address the following specific aims:
Aim 1 will determine the role of epicardial 2-catenin in cell fate specification and maturation of epicardial derivatives ex vivo and in vivo using time-lapse analysis of explanted tissues, lineage tracing, and mechanistic studies in epicardial-2-catenin mutant mice.
In Aim 2 we will determine whether Wnt activity regulates the formation of the coronary arteries and the expansion of the compact zone. To address this question, we will use a genetic strategy to block 2-catenin nuclear translocation and analyze signaling downstream of Wnt in the epicardium.. Finally, in Aim 3 we will determine whether 2 catenin is a functional mediator of RXR1 signaling. As a proof of principle for this hypothesis, we propose the rescue of the RXR1 phenotype by interbreeding the RXR1 mutant mice with the conditional 2 catenin/loxP (ex3) mouse, resulting in mice with constitutively active 2 catenin in the epicardium. Completion of this proposal will unravel the mechanisms of retinoid/2 catenin signaling in coronary formation and ventricular compaction and might offer the potential for therapeutic interventions to ameliorate or even treat coronary disorders that are leading causes of mortality.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL065484-10
Application #
8228081
Study Section
Cardiovascular Differentiation and Development Study Section (CDD)
Program Officer
Schramm, Charlene A
Project Start
2000-07-01
Project End
2013-12-31
Budget Start
2012-01-01
Budget End
2013-12-31
Support Year
10
Fiscal Year
2012
Total Cost
$393,512
Indirect Cost
$146,012
Name
Stanford University
Department
Pediatrics
Type
Schools of Medicine
DUNS #
009214214
City
Stanford
State
CA
Country
United States
Zip Code
94305
Mercola, Mark; Ruiz-Lozano, Pilar; Schneider, Michael D (2011) Cardiac muscle regeneration: lessons from development. Genes Dev 25:299-309
Prigozhina, Natalie L; Heisel, Andrew; Wei, Ke et al. (2011) Characterization of a novel angiogenic model based on stable, fluorescently labelled endothelial cell lines amenable to scale-up for high content screening. Biol Cell 103:467-81
Sridurongrit, Somyoth; Larsson, Jonas; Schwartz, Robert et al. (2008) Signaling via the Tgf-beta type I receptor Alk5 in heart development. Dev Biol 322:208-18
St Amand, Tara R; Lu, Jonathan T; Zamora, Monica et al. (2006) Distinct roles of HF-1b/Sp4 in ventricular and neural crest cells lineages affect cardiac conduction system development. Dev Biol 291:208-17
Kaliman, Perla; Catalucci, Daniele; Lam, Jason T et al. (2005) Myotonic dystrophy protein kinase phosphorylates phospholamban and regulates calcium uptake in cardiomyocyte sarcoplasmic reticulum. J Biol Chem 280:8016-21