Ischemic cardiomyopathy (ICM) is the most prevalent type of chronic heart diseases in the United States and a common underlying etiology of congestive heart failure. Nonetheless the underlying molecular mechanism of ICM remains unclear. Moreover, the prognosis of chronic ICM is very poor if left untreated. Therefore it is highly important to gain a better understanding of the disease mechanism and to identify specific biomarkers for early detection and treatment of ICM. Myofilament proteins in the sarcomeres not only play essential roles in cardiac contractility but also are critical elements in signal reception and transduction during the onset and progression to heart failure. The hypothesis is that post-myocardial infarction (MI) maladaptive cardiac remodeling can result in altered protein modifications in myofilaments that are associated with cardiac dysfunction and offer potential diagnostic and prognostic biomarkers of ICM. Although multiple myofilament post-translational modifications (PTMs) are believed to act in concert in regulating cardiac function, a comprehensive analysis of myofilament proteins simultaneously and assess all synergistic PTM changes related to cardiac function is lacking. Herein, we will employ an innovative integrated """"""""top-down"""""""" protein mass spectrometry (MS)-based disease proteomics platform to simultaneously examine myofilament proteins extracted from healthy and diseased swine and human myocardium and identify all disease-related changes in myofilament modifications.
Aim 1 will identify altered protein modifications in myofilaments from diseased swine myocardium of post-MI ICM model and determine their functional consequences.
Aim 2 will determine myofilament alterations in end-stage failing human myocardium from ICM patients and their functional consequences.
Aim 3 will assess the functional effects of novel myofilament modifications in Ca2+-mediated cardiac muscle contraction and relaxation. Mechanical properties (the isometric force and the Ca2+-sensitivity of the force) will be measured in skinned myocardial preparations in parallel to proteomics experiments. Furthermore, we will assess regional systolic and diastolic function to establish mechanistic links between specific proteomic changes and in vivo and ex vivo cardiac function. With the completion of this project, we expect to identify multiple functionally significant alterations in myofilaments and understand how these alterations act in concert to modulate maladaptive signaling during post-MI left ventricular remodeling to failure. The success of our research will provide new insights into the mechanisms underlying ICM and will identify new candidate biomarkers for early detection of the presence and progression of ICM.

Public Health Relevance

Heart failure remains a leading cause of mortality and morbidity in the United States. Whereas the treatments for acute coronary syndromes have dramatically improved in the past decade, chronic heart disease is approaching epidemic levels in western countries. The success of our research will provide new insights into the mechanisms underlying chronic heart disease and will also identify new candidate biomarkers for early detection of the presence and progression of Ischemic cardiomyopathy.

National Institute of Health (NIH)
National Heart, Lung, and Blood Institute (NHLBI)
Research Project (R01)
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Myocardial Ischemia and Metabolism Study Section (MIM)
Program Officer
Schwartz, Lisa
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University of Wisconsin Madison
Anatomy/Cell Biology
Schools of Medicine
United States
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Chen, Bifan; Brown, Kyle A; Lin, Ziqing et al. (2018) Top-Down Proteomics: Ready for Prime Time? Anal Chem 90:110-127
Smelter, Dan F; de Lange, Willem J; Cai, Wenxuan et al. (2018) The HCM-linked W792R mutation in cardiac myosin-binding protein C reduces C6 FnIII domain stability. Am J Physiol Heart Circ Physiol 314:H1179-H1191
Cai, Wenxuan; Hite, Zachary L; Lyu, Beini et al. (2018) Temperature-sensitive sarcomeric protein post-translational modifications revealed by top-down proteomics. J Mol Cell Cardiol 122:11-22
Lin, Ziqing; Guo, Fang; Gregorich, Zachery R et al. (2018) Comprehensive Characterization of Swine Cardiac Troponin T Proteoforms by Top-Down Mass Spectrometry. J Am Soc Mass Spectrom 29:1284-1294
Wu, Zhijie; Tiambeng, Timothy N; Cai, Wenxuan et al. (2018) Impact of Phosphorylation on the Mass Spectrometry Quantification of Intact Phosphoproteins. Anal Chem 90:4935-4939
Gao, Ling; Gregorich, Zachery R; Zhu, Wuqiang et al. (2018) Large Cardiac Muscle Patches Engineered From Human Induced-Pluripotent Stem Cell-Derived Cardiac Cells Improve Recovery From Myocardial Infarction in Swine. Circulation 137:1712-1730
Lam, Maggie P Y; Ge, Ying (2018) Harnessing the Power of Proteomics to Assess Drug Safety and Guide Clinical Trials. Circulation 137:1011-1014
Martin-Garrido, Abel; Biesiadecki, Brandon J; Salhi, Hussam E et al. (2018) Monophosphorylation of cardiac troponin-I at Ser-23/24 is sufficient to regulate cardiac myofibrillar Ca2+ sensitivity and calpain-induced proteolysis. J Biol Chem 293:8588-8599
Chen, Zhilong; Song, Jiangping; Chen, Liang et al. (2018) Characterization of TTN Novex Splicing Variants across Species and the Role of RBM20 in Novex-Specific Exon Splicing. Genes (Basel) 9:
Chen, Bifan; Lin, Ziqing; Alpert, Andrew J et al. (2018) Online Hydrophobic Interaction Chromatography-Mass Spectrometry for the Analysis of Intact Monoclonal Antibodies. Anal Chem 90:7135-7138

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