Although recent studies demonstrate that humans can generate new heart cells, the rate of renewal dramatically decreases with age. Since heart failure often strikes in the second half of life, understanding how heart cells regenerate and why this process declines with age is a critical goal. The inducible cre-lox approach is a widely-used method for fate-mapping of cells in mammals. In this proposal, we show preliminary data revealing that an inducible cre-lox approach to genetically label cardiomyocytes in mice is not suitable for studying the aging process in the myocardium. Our preliminary data reveal age-related recombination in cardiomyocytes in the absence of induction of the cre activity. Thus, we have developed an entirely new approach to fate-mapping using isotopes in a cell-specific metabolic precursor. This metabolic fate-mapping approach utilizes an isotope-enriched metabolic tracer, specifically creatine, which is taken up by cells and utilized in the cytoplasmic phosphocreatine shuttle. The intracellular creatine pool is known to turn over at a rate of approximately 2-3% per day, making it an ideal cardiomyocyte-specific metabolic label. Cells that are creatine positive can be identified via use of Multi-Isotope Imaging Mass Spectrometry (MIMS), a high resolution quantitative approach. Using this new metabolic fate-mapping technique, the specific aims for this study are: 1. To test the hypothesis that, together with MIMS, metabolic fate-mapping using isotope-labeled creatine will enable the detection of cardiomyocyte regeneration during normal aging for mice of all age groups. We anticipate that the rate of regeneration will be low, and this basal cell division rate will decrease as a function of age. 2. To test the hypothesis that, together with MIMS, metabolic fate-mapping using isotope-labeled creatine will enable the detection of cardiomyocyte regeneration after injury for mice of all age groups. We anticipate that regeneration will primarily be from stem cell differentiation and not by pre-existing cardiomyocyte cell division, and this rate of regeneration will decrease as a function of age. This metabolic fate-mapping approach will enable the study of aging-related regeneration of cardiomyocytes. Furthermore, this new approach can be used not only for the myocardium but in other types of tissues such as skeletal muscle, brain and adipocytes. Finally, because this approach can be applied with stable, non-radioactive isotopes, long-term clinical studies of regenerative activity in human subjects may be enabled by this method.

Public Health Relevance

Heart Failure, a leading cause of death and disability in the United States, is often due to the loss of heart muscle cells, with inadequate replacement by new heart cells. Understanding why heart cells don't regenerate in older age is a critical goal of cardiovascular science. This project will develop a new approach to measuring birth of new heart cells, allowing us to understand the effect of aging on the heart.

Agency
National Institute of Health (NIH)
Institute
National Institute on Aging (NIA)
Type
Research Project (R01)
Project #
5R01AG040019-03
Application #
8464620
Study Section
Special Emphasis Panel (ZAG1-ZIJ-2 (M1))
Program Officer
Kohanski, Ronald A
Project Start
2011-08-15
Project End
2016-07-31
Budget Start
2013-08-01
Budget End
2014-07-31
Support Year
3
Fiscal Year
2013
Total Cost
$359,506
Indirect Cost
$146,881
Name
Brigham and Women's Hospital
Department
Type
DUNS #
030811269
City
Boston
State
MA
Country
United States
Zip Code
02115
Mahmoud, Ahmed I; Lee, Richard T (2016) Adrenergic function restoration in the transplanted heart: a role for neural crest-derived cells. Cardiovasc Res 109:348-9
Walker, Ryan G; Poggioli, Tommaso; Katsimpardi, Lida et al. (2016) Biochemistry and Biology of GDF11 and Myostatin: Similarities, Differences, and Questions for Future Investigation. Circ Res 118:1125-41; discussion 1142
Poggioli, Tommaso; Vujic, Ana; Yang, Peiguo et al. (2016) Circulating Growth Differentiation Factor 11/8 Levels Decline With Age. Circ Res 118:29-37
Uygur, Aysu; Lee, Richard T (2016) Mechanisms of Cardiac Regeneration. Dev Cell 36:362-74
Dotimas, James R; Lee, Austin W; Schmider, Angela B et al. (2016) Diabetes regulates fructose absorption through thioredoxin-interacting protein. Elife 5:
O'Meara, Caitlin C; Lee, Richard T (2015) Peering Into the Cardiomyocyte Nuclear Epigenetic State. Circ Res 117:392-4
O'Meara, Caitlin C; Wamstad, Joseph A; Gladstone, Rachel A et al. (2015) Transcriptional reversion of cardiac myocyte fate during mammalian cardiac regeneration. Circ Res 116:804-15
Mahmoud, Ahmed I; O'Meara, Caitlin C; Gemberling, Matthew et al. (2015) Nerves Regulate Cardiomyocyte Proliferation and Heart Regeneration. Dev Cell 34:387-99
Jiang, Jianming; Burgon, Patrick G; Wakimoto, Hiroko et al. (2015) Cardiac myosin binding protein C regulates postnatal myocyte cytokinesis. Proc Natl Acad Sci U S A 112:9046-51
Bryant, Donald Marion; O'Meara, Caitlin Claire; Ho, Nhi Ngoc et al. (2015) A systematic analysis of neonatal mouse heart regeneration after apical resection. J Mol Cell Cardiol 79:315-8

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