AMP-activated protein kinase (AMPK) is serine/threonine kinase that serves as an intracellular energy sensor and a key regulator of cell metabolism, growth and death, hence a promising drug target for obesity, diabetes, cancer and cardiovascular diseases. AMPK is a heterotrimeric complex composed of catalytic 1-subunit and regulatory 2- and 3-subunits with multiple isoforms for each subunit. Previous studies have shown that activation of AMPK in the heart during stress is an important cardioprotective mechanism. However, point mutations in the regulatory 32-subunit (encoded by prkag2 gene) of AMPK cause human cardiomyopathy characterized by cardiac hypertrophy, arrhythmias and glycogen storage. This raises concerns of future drug development targeting AMPK cascade in the heart. It is thus critical to understand the disease mechanisms responsible for the prkag2 cardiomyopathy. Using transgenic mice overexpressing mutant prkag2 (N488I) in the heart (TG32N488I) that faithfully recapitulated prkag2 cardiomyopathy, we in the previous funding period demonstrated that the mutation caused aberrant activation of AMPK in the absence of energetic deficit, and the cardiomyopathy phenotype could be rescued by inhibition of AMPK activity. We have further demonstrated that the simultaneous increases in glucose uptake and fatty acid oxidation in the absence of energy stress resulted in metabolic re-routing of exogenous glucose preferentially into glycogen pool in the TG32N488I hearts. To determine whether the cardiomyopathy phenotype is entirely attributable to glycogen storage, we sought to rescue the glycogen storage without changing the mutant gene expression by specifically targeting the activity of muscle glycogen synthase. Our preliminary data showed that this strategy successfully normalized cardiac glycogen content but not the cardiac hypertrophy phenotype in TG32N488I mice. Thus, we hypothesize that 32- AMPK mutation causes cardiac hypertrophy independent of glycogen storage. Moreover, since the prkag2 mutation causes predominantly cardiac phenotype, it raises the question of a unique role of 32-AMPK in the heart. This led us to generate mouse models deficient of 32-AMPK in order to determine the isoform-specific function of 3-AMPK. Using these models we propose 1) to test the hypothesis that the prkag2 mutation causes cardiac hypertrophy independent of glycogen storage and, to determine the mechanisms by which aberrant activation of 32-AMPK under nutrient saturated conditions stimulate cardiac hypertrophy;2) to define the composition and the function of the 32-AMPK signaling cascade in the heart during cardiac development and diseases.

Public Health Relevance

This project investigates disease mechanisms of human cardiomyopathy caused by mutations of the gene encoding g2-subunit of the AMP-activated protein kinase (AMPK). Because AMPK is a promising drug target for metabolic disorders, cardiovascular diseases and cancer, this project will also define the signaling mechanisms of g2-AMPK pathway in the heart in order to provide a basis for novel therapeutic strategies.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL088634-06
Application #
8389882
Study Section
Myocardial Ischemia and Metabolism Study Section (MIM)
Program Officer
Wong, Renee P
Project Start
2007-05-15
Project End
2015-11-30
Budget Start
2012-12-01
Budget End
2013-11-30
Support Year
6
Fiscal Year
2013
Total Cost
$501,198
Indirect Cost
$218,203
Name
University of Washington
Department
Anesthesiology
Type
Schools of Medicine
DUNS #
605799469
City
Seattle
State
WA
Country
United States
Zip Code
98195
Kim, Maengjo; Hunter, Roger W; Garcia-Menendez, Lorena et al. (2014) Mutation in the ?2-subunit of AMP-activated protein kinase stimulates cardiomyocyte proliferation and hypertrophy independent of glycogen storage. Circ Res 114:966-75
Yu, Qiujun; Lee, Chi Fung; Wang, Wang et al. (2014) Elimination of NADPH oxidase activity promotes reductive stress and sensitizes the heart to ischemic injury. J Am Heart Assoc 3:e000555
Kolwicz Jr, Stephen C; Purohit, Suneet; Tian, Rong (2013) Cardiac metabolism and its interactions with contraction, growth, and survival of cardiomyocytes. Circ Res 113:603-16
Garcia-Menendez, Lorena; Karamanlidis, Georgios; Kolwicz, Stephen et al. (2013) Substrain specific response to cardiac pressure overload in C57BL/6 mice. Am J Physiol Heart Circ Physiol 305:H397-402
Karamanlidis, Georgios; Lee, Chi Fung; Garcia-Menendez, Lorena et al. (2013) Mitochondrial complex I deficiency increases protein acetylation and accelerates heart failure. Cell Metab 18:239-50
Kim, Maengjo; Shen, Mei; Ngoy, Soeun et al. (2012) AMPK isoform expression in the normal and failing hearts. J Mol Cell Cardiol 52:1066-73
Kim, Maengjo; Tian, Rong (2011) Targeting AMPK for cardiac protection: opportunities and challenges. J Mol Cell Cardiol 51:548-53
Kolwicz Jr, Stephen C; Tian, Rong (2011) Glucose metabolism and cardiac hypertrophy. Cardiovasc Res 90:194-201
Karamanlidis, Georgios; Nascimben, Luigino; Couper, Gregory S et al. (2010) Defective DNA replication impairs mitochondrial biogenesis in human failing hearts. Circ Res 106:1541-8
Kolwicz Jr, Stephen C; Tian, Rong (2010) Assessment of cardiac function and energetics in isolated mouse hearts using 31P NMR spectroscopy. J Vis Exp :

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