In this application we propose to develop a preventive gene therapy strategy for myocardium protection from future ischemia/reperfusion injury (I/R) involving a single administration of a therapeutic gene with a vector system capable of efficient and long term myocyte specific and inducible expression of the therapeutic gene. We will employ recombinant adeno-associated viral vectors (rAAV) to achieve long-term and stable expression of transduced genes in the myocardium. We will select cis-acting promoter elements that are capable of conferring inducible and cell-specific gene expression and to use these elements as molecular switches to achieve optimal temporal and spatial control of expression of heme oxygenase-1 (HO-1) as the therapeutic transgene. We have reported that transgenic mice with cardiac-directed overexpression of HO-1 develop resistance to I/R-induced myocardial injury; furthermore, a single intramyocardial delivery of HO-1 gene by rAAV in rats, eight weeks in advance of I/R-induced myocardial injury, resulted in dramatic reduction in myocardial infarction, demonstrating that the combination of rAAV vector and HO-1 gene is an efficacious and feasible therapeutic strategy for myocardial protection. Accordingly, in this proposal we will (1) first document the impact of rAAV-mediated intramyocardial delivery of HO-1 on long-term survival following acute myocardial infarction induced by I/R injury and to determine the duration of expression and sustainability of the therapeutic effect of the transgene; (2) we will develop inducible, hypoxia and reoxygenation-sensitive expression of HO-1 as an endogenously regulated therapeutic approach for myocardial protection; (3) We will establish cardiomyocyte specific transgene expression of HO-1, and finally (4) we will develop a combined approach of cell-specific and inducible expression as an ideal strategy for transgene delivery with maximal tissue specificity and safety.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
7R01HL072010-03
Application #
6986621
Study Section
Cardiovascular and Renal Study Section (CVB)
Project Start
2003-05-02
Project End
2007-03-31
Budget Start
2004-10-01
Budget End
2005-03-31
Support Year
3
Fiscal Year
2004
Total Cost
$17,300
Indirect Cost
Name
Duke University
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
044387793
City
Durham
State
NC
Country
United States
Zip Code
27705
Hodgkinson, Conrad P; Gomez, José A; Baksh, Syeda Samara et al. (2018) Insights from molecular signature of in vivo cardiac c-Kit(+) cells following cardiac injury and ?-catenin inhibition. J Mol Cell Cardiol 123:64-74
Dal-Pra, Sophie; Hodgkinson, Conrad P; Mirotsou, Maria et al. (2017) Demethylation of H3K27 Is Essential for the Induction of Direct Cardiac Reprogramming by miR Combo. Circ Res 120:1403-1413
Matsushita, Kenichi; Wu, Yaojiong; Pratt, Richard E et al. (2016) Deletion of angiotensin II type 2 receptor accelerates adipogenesis in murine mesenchymal stem cells via Wnt10b/beta-catenin signaling. Lab Invest 96:909-17
Li, Yanzhen; Dal-Pra, Sophie; Mirotsou, Maria et al. (2016) Tissue-engineered 3-dimensional (3D) microenvironment enhances the direct reprogramming of fibroblasts into cardiomyocytes by microRNAs. Sci Rep 6:38815
Matsushita, Kenichi; Morello, Fulvio; Zhang, Zhiping et al. (2016) Nuclear hormone receptor LXR? inhibits adipocyte differentiation of mesenchymal stem cells with Wnt/beta-catenin signaling. Lab Invest 96:230-8
Hodgkinson, Conrad P; Bareja, Akshay; Gomez, José A et al. (2016) Emerging Concepts in Paracrine Mechanisms in Regenerative Cardiovascular Medicine and Biology. Circ Res 118:95-107
Yuan, Hsiangkuo; Gomez, Jose A; Chien, Jennifer S et al. (2016) Tracking mesenchymal stromal cells using an ultra-bright TAT-functionalized plasmonic-active nanoplatform. J Biophotonics 9:406-13
Yang, Yanqiang; Gomez, Jose A; Herrera, Marcela et al. (2015) Salt restriction leads to activation of adult renal mesenchymal stromal cell-like cells via prostaglandin E2 and E-prostanoid receptor 4. Hypertension 65:1047-54
Jayawardena, Tilanthi M; Finch, Elizabeth A; Zhang, Lunan et al. (2015) MicroRNA induced cardiac reprogramming in vivo: evidence for mature cardiac myocytes and improved cardiac function. Circ Res 116:418-24
Hodgkinson, Conrad P; Kang, Martin H; Dal-Pra, Sophie et al. (2015) MicroRNAs and Cardiac Regeneration. Circ Res 116:1700-11

Showing the most recent 10 out of 50 publications