Bmx Tyrosine Kinase Signaling in Cardiac Protection

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DESCRIPTION (provided by applicant): Ischemic preconditioning is the most potent and consistently reproducible method of preventing heart tissue from undergoing irreversible ischemic damage. Induction of this phenomenon can be recapitulated in lieu of ischemia by drugs such as nitric oxide (NO) donors, which significantly reduce myocardial infarct size. Cardiac cell death in ischemia/reperfusion injury is known to centrally involve malfunction of the mitochondria (mitochondrial permeability transition [MPT]), and protection thus requires prevention of MPT. Although the signaling mechanisms which prevent mitochondrial dysfunction in cardiac protection are unknown, tyrosine kinase signaling has been implicated based on the ability of tyrosine kinase inhibitors to block cardiac protection. The non-receptor tyrosine kinase, Bmx, which has an established role in cytoprotective signaling in non-cardiac cells, was recently identified in cardiac tissue. This protein has been shown to be activated by PI3-K and Src in non-cardiac cells, although how it is activated in the cardiomyocyte is completely unknown. Our preliminary results have shown that the hearts of Bmx knockout (KO) mice cannot be protected with NO donors, thereby implicating Bmx as a previously unrecognized necessary component of protective signaling. Because of these observations, we hypothesize that Bmx signaling is activated in cardiomyocytes in response to cardiac protection and promotes cell survival, at least in part, by regulating mitochondrial function. We propose two Aims that combine biochemistry, proteomic analysis, confocal microscopy and cell/organelle physiology to test this hypothesis: First, we will characterize Bmx activation in mouse cardiomyocytes during NO donor induced protection by determining the role of PI3-K and Src, identifying activators of Bmx and mapping the phosphorylation sites necessary for cardiac protective signaling. Second, we will evaluate the role of Bmx signaling in NO donor induced cardiac protection by measuring mitochondrial function in Bmx KO mice, identifying Bmx associated proteins at this organelle, and determining Bmx-dependent changes in mitochondrial protein expression. Determining Bmx signaling mechanisms will establish the first insights into a newly discovered family of tyrosine kinases in the heart and allow a better understanding of the protective phenotype with the goal of developing molecular approaches to treat ischemic heart disease.