? Development and application of non-invasive imaging techniques to physically and functionally assess the potential of novel cardiomyocyte stem cells to regenerate damaged myocardium have clear clinical significance. Repair and replacement of damaged cardiac muscle tissue is a key limitation in the recovery from ischemic and other forms of heart disease, the leading cause of death in the United States. New strategies for regenerating damaged cardiac muscle are urgently needed and could be readily translated to the clinical setting if first shown to be effective in animal models. The proposal outlined here will demonstrate the feasibility of using molecular imaging techniques in vivo to evaluate our novel stem cell-based strategy for cardiac regenerative medicine. This research proposal directly addresses both of the stated goals of the initiative outlined in RFA-HL-04-003 (Cellular and Molecular Imaging of the Cardiovascular, Pulmonary, and Hematopoetic Systems): (1) Detect and quantify molecular and cellular pathways that regulate heart function...(we describe and will further characterize a novel cardiomyocyte differentiation pathway), and (2) Develop new methods for cell tracking in vivo for applications in cell-based therapeutics...(we will transplant novel cardiomyocyte stem cells into damaged hearts and track them in vivo using advanced magnetic resonance and bioluminescent imaging methods). Our data indicate that cardiac cells transiently expressing the enzyme, Phenylethanolamine n-methyltransferase (Pnmt), ultimately become myocytes that contribute substantially to pacemaking, conduction, and working myocardium. Thus, Pnmt serves as a novel marker of cardiomyocyte stem cells (Pnmt+ cells) in the developing heart. We will test the idea that Pnmt+ cells can be selectively isolated and transplanted into damaged cardiac regions where they will be monitored both physically (location) and physiologically (function) using non-invasive molecular and cellular imaging techniques to evaluate the potential of these cells to regenerate cardiac muscle tissue in vivo. ? Three specific aims are proposed to accomplish this goal: ? Aim 1: Isolate and characterize Pnmt-nEGFP+ cells from cardiac-differentiated embryonic stem cells.
Aim 2 : In vivo magnetic resonance imaging (MRI) of transplanted cardiomyocyte stem (Pnmt+) cells.
Aim 3 : In vivo bioluminescence imaging (BLI) of transplanted cardiomyocyte stem (Pnmt+) cells. ? (End of Abstract) ? ?

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
3R01HL078716-03S1
Application #
7287063
Study Section
Special Emphasis Panel (ZHL1)
Program Officer
Buxton, Denis B
Project Start
2004-09-22
Project End
2008-08-31
Budget Start
2006-09-01
Budget End
2007-08-31
Support Year
3
Fiscal Year
2006
Total Cost
$36,920
Indirect Cost
Name
University of Central Florida
Department
Other Basic Sciences
Type
Schools of Medicine
DUNS #
150805653
City
Orlando
State
FL
Country
United States
Zip Code
32826
Baker, Candice N; Gidus, Sarah A; Price, George F et al. (2015) Impaired cardiac energy metabolism in embryos lacking adrenergic stimulation. Am J Physiol Endocrinol Metab 308:E402-13
Osuala, Kingsley; Baker, Candice N; Nguyen, Ha-Long et al. (2012) Physiological and genomic consequences of adrenergic deficiency during embryonic/fetal development in mice: impact on retinoic acid metabolism. Physiol Genomics 44:934-47
Baker, Candice; Taylor, David G; Osuala, Kingsley et al. (2012) Adrenergic deficiency leads to impaired electrical conduction and increased arrhythmic potential in the embryonic mouse heart. Biochem Biophys Res Commun 423:536-41
Xia, Jixiang; Martinez, Angela; Daniell, Henry et al. (2011) Evaluation of biolistic gene transfer methods in vivo using non-invasive bioluminescent imaging techniques. BMC Biotechnol 11:62
Osuala, Kingsley; Telusma, Kathleen; Khan, Saad M et al. (2011) Distinctive left-sided distribution of adrenergic-derived cells in the adult mouse heart. PLoS One 6:e22811
Kammili, Ramana K; Taylor, David G; Xia, Jixiang et al. (2010) Generation of novel reporter stem cells and their application for molecular imaging of cardiac-differentiated stem cells in vivo. Stem Cells Dev 19:1437-48
Ebert, Steven N; Rong, Qi; Boe, Steve et al. (2008) Catecholamine-synthesizing cells in the embryonic mouse heart. Ann N Y Acad Sci 1148:317-24
Ebert, Steven N; Taylor, David G; Nguyen, Ha-Long et al. (2007) Noninvasive tracking of cardiac embryonic stem cells in vivo using magnetic resonance imaging techniques. Stem Cells 25:2936-44
Knollmann, Bjorn C; Sirenko, Syevda; Rong, Qi et al. (2007) Kcnq1 contributes to an adrenergic-sensitive steady-state K+ current in mouse heart. Biochem Biophys Res Commun 360:212-8
Ebert, Steven N; Taylor, David G (2006) Catecholamines and development of cardiac pacemaking: an intrinsically intimate relationship. Cardiovasc Res 72:364-74