The long-term objective of this proposal is to establish a unified theory to describe the mechanisms of crosstalk signaling between Ca2+ and reactive oxygen species (ROS) in cardiac muscle cells, and to translate these signaling mechanisms to the physiology and pathology of cardiac function. The pivotal role of mitochondrial Ca2+ and ROS in mediating the life and death of cardiac muscle cells is well recognized. The separation between mitochondria-mediated life versus death resides in the finest balance between concentrations of Ca2+ and ROS. The majority of existing research in the field focuses individually either on Ca2+ or ROS homeostasis. The interaction between these two signaling pathways, however, has just begun to gain attention by a small number of laboratories. Intriguingly, there is a growing library of literature suggesting that mitochondrial dynamics (fission, fusion, and trafficking) play an essential role in the physiological regulation of cellular ATP, Ca2+, and ROS homeostasis. In this proposal, we will study how these three important components (Ca2+, ROS, and mitochondrial fission machinery) communicate to regulate cardiac Ca2+ and ROS crosstalk signaling. We will use a multidisciplinary approach encompassing techniques of cell biology (e.g. confocal microscopy), molecular biology (e.g. gene transfer), biochemistry (e.g. western blots), and transgenic mouse models (e.g. cyclophilin D (CypD) knockout mice and mt-cpYFP transgenic mice) to elucidate the mechanisms of ROS and Ca2+ symbiosis with an unique emphasis on mitochondrial fission protein DLP1 and mitochondrial permeability transition (MPT). Our central hypothesis is: an increased mitochondrial Ca2+ concentration ([Ca2+]m) favors the balance of mitochondrial dynamics towards fission that in turn increases ROS generation. The resulting oxidized environment leads to additional mitochondrial Ca2+ influx. Both the increases in [Ca2+]m and ROS enhance the opening probability of MPT that further augments ROS generation. Eventually, this positive feedback loop is counter balanced by Ca2+ and ROS activated mitochondrial Ca2+ efflux mechanisms including Na/Ca exchange and MPT. The three specific aims are: 1) To determine whether an increased [Ca2+]m promotes mitochondrial fission processes, which then lead to increase ROS generation. 2) To assess the contribution of a CypD containing MPT pathway in [Ca2+]m-mediated ROS generation. 3) To determine the role of MPT as a rapid Ca2+ efflux mechanism of mitochondria.

Public Health Relevance

Oxidative stress can cause numerous human diseases including cardiomyopathy in chronic heart failure, ischemic heart disease, neurodegenerative diseases, diabetes, obesity, and aging. The proposed research focuses on the elucidation of cellular mechanisms of reactive oxygen species generation. It is our objective, not only to make a scientific contribution to oxidative stress-mediated disease mechanisms, but also to develop possible therapeutic means for treating these debilitating disorders.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
1R01HL093671-01A1
Application #
7805152
Study Section
Electrical Signaling, Ion Transport, and Arrhythmias Study Section (ESTA)
Program Officer
Przywara, Dennis
Project Start
2010-01-01
Project End
2011-01-31
Budget Start
2010-01-01
Budget End
2011-01-31
Support Year
1
Fiscal Year
2010
Total Cost
$379,083
Indirect Cost
Name
University of Rochester
Department
Pharmacology
Type
Schools of Dentistry
DUNS #
041294109
City
Rochester
State
NY
Country
United States
Zip Code
14627
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Mishra, Jyotsna; Jhun, Bong Sook; Hurst, Stephen et al. (2017) The Mitochondrial Ca2+ Uniporter: Structure, Function, and Pharmacology. Handb Exp Pharmacol 240:129-156
Zhang, Huiliang; Wang, Pei; Bisetto, Sara et al. (2017) A novel fission-independent role of dynamin-related protein 1 in cardiac mitochondrial respiration. Cardiovasc Res 113:160-170
Wang, Wang; Fernandez-Sanz, Celia; Sheu, Shey-Shing (2017) Regulation of mitochondrial bioenergetics by the non-canonical roles of mitochondrial dynamics proteins in the heart. Biochim Biophys Acta :
Wang, Wang; Gong, Guohua; Wang, Xianhua et al. (2016) Mitochondrial Flash: Integrative Reactive Oxygen Species and pH Signals in Cell and Organelle Biology. Antioxid Redox Signal 25:534-49
Jhun, Bong Sook; Mishra, Jyotsna; Monaco, Sarah et al. (2016) The mitochondrial Ca2+ uniporter: regulation by auxiliary subunits and signal transduction pathways. Am J Physiol Cell Physiol 311:C67-80
O-Uchi, Jin; Sorenson, Jaime; Jhun, Bong Sook et al. (2015) Isoform-specific dynamic translocation of PKC by ?1-adrenoceptor stimulation in live cells. Biochem Biophys Res Commun 465:464-70
Fujiwara, Keigi; Sheu, Shey-Shing (2015) Mitochondrial dynamics regulate neointima formation. Cardiovasc Res 106:175-7
Gomez, L; Thiebaut, P-A; Paillard, M et al. (2015) The SR/ER-mitochondria calcium crosstalk is regulated by GSK3? during reperfusion injury. Cell Death Differ 22:1890
Pan, Shi; Wang, Nadan; Bisetto, Sara et al. (2015) Downregulation of adenine nucleotide translocator 1 exacerbates tumor necrosis factor-?-mediated cardiac inflammatory responses. Am J Physiol Heart Circ Physiol 308:H39-48

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