Clin Res Cardiol (2022). https://doi.org/10.1007/s00392-022-02002-5 |
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Unbiased screen of the cardiac proteome in a rat HFpEF model reveals key pathway changes due to obesity | ||
F. Koser1, M. Abdellatif2, C. Türk3, A. J. Hobbach4, H. Reinecke4, M. Krüger3, S. Sedej2, W. A. Linke1 | ||
1Institute of Physiology II, University Hospital Münster, Münster; 2Division of Cardiology, Medical University of Graz, Graz, Österreich; 3Institute for Genetics, University of Cologne, Cologne; 4Department of Cardiology I, University Hospital Münster, Münster; | ||
Background: Despite the high public-health burden, the pathomechanism of heart failure with preserved ejection fraction (HFpEF) remains elusive and evidence-based therapies are lacking. Diastolic dysfunction is a key feature in HFpEF patients, caused in part by increased stiffness of the cardiomyocyte protein titin. The current paradigm of HFpEF pathophysiology postulates meta-inflammation and oxidative stress as crucial disease triggers, resulting from a combination of metabolic syndrome and hypertension. Good patient-mimicking animal models of HFpEF are sparse but one of the few established models is the Zucker diabetic fatty/Spontaneous hypertensive heart failure F1 hybrid (ZSF1) rat, which does have hypertension and metabolic syndrome. The ZSF1 rat is available as a lean (no HFpEF) and an obese variant (HFpEF), which have almost the same genetic background. Objective: To test how the presence of obesity alters protein expression and function in the hearts of ZSF1-obese versus ZSF1-lean rats, and to gain new insights into the HFpEF pathomechanism using unbiased mass spectrometry-based proteomic screens, as well as immunoblotting. Methods & Results: Eight-week-old female and male ZSF1-obese and ZSF1-lean rats were fed with a Purina diet until the onset of the HFpEF phenotype in the ZSF1-obese animals at around 20 weeks of age. Mass spectrometric analyses of heart tissue samples showed that the cardiac proteome and phosphoproteome in ZSF1-obese versus ZSF1-lean were less markedly altered than the acetylome. Many acetylation changes affected proteins involved in cardiac metabolism and energy production, suggesting obesity as a crucial comorbidity in the HFpEF pathogenesis. Some of the proposed key features of the HFpEF pathomechanism could be confirmed in ZSF1-obese rats by immunoblot-based quantitation, while others could not. Unexpectedly, both eNos activity and iNos expression (markers of oxidative/nitrosative stress) were unaltered in ZSF1-obese hearts, as was the phospho-Ire1α:Ire1α ratio (indicating Ire1α activity), a measure of the unfolded protein response pathway. In contrast, CD68 and P-selectin expression were increased in ZSF1-obese versus ZSF1-lean, indicating macrophage infiltration and microvascular endothelial inflammation. Moreover, PKG activity and PKG site-specific titin phosphorylation in the N2Bus segment were decreased in the hearts of ZSF1-obese rats, consistent with increased titin-based cardiomyocyte stiffness. In addition, titin showed several hypo-acetylated sites in ZSF1-obese versus ZSF1-lean hearts, which could also alter titin-based stiffness. Disturbed Ca2+-handling in ZSF1-obese hearts was evidenced by altered posttranslational modifications of the ryanodine receptor (hyper-phosphorylated) and SERCA (hyper-acetylated). Conclusion: Our findings suggest that obesity on the background of hypertension and additional co-morbidities triggers crucial proteomic and posttranslational alterations in HFpEF hearts, which lead to meta-inflammation and cardiomyocyte stiffening. Altered acetylation of multiple cardiac proteins caused by obesity could be a key posttranslational modification in HFpEF. |
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https://dgk.org/kongress_programme/jt2022/aP816.html |