Secretome Analysis of Cardiomyocytes Identifies PCSK6 (Proprotein Convertase Subtilisin/Kexin Type 6) as a Novel Player in Cardiac Remodeling After Myocardial Infarction
| العنوان: | Secretome Analysis of Cardiomyocytes Identifies PCSK6 (Proprotein Convertase Subtilisin/Kexin Type 6) as a Novel Player in Cardiac Remodeling After Myocardial Infarction |
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| المؤلفون: | Xue Li, Johannes Backs, Felix Lasitschka, Hugo A. Katus, Florian Sicklinger, Tim Christian Kuhn, Andreas Jungmann, Florian Leuschner, Jeroen Krijgsveld, Oliver J. Müller, Ingmar Sören Meyer, Sonja Burkert-Rettenmaier, Johannes Knobel |
| المصدر: | Circulation |
| بيانات النشر: | Ovid Technologies (Wolters Kluwer Health), 2020. |
| سنة النشر: | 2020 |
| مصطلحات موضوعية: | Male, medicine.medical_specialty, Proteome, Myocardial Infarction, Acute occlusion, 030204 cardiovascular system & hematology, Ventricular Function, Left, Article, 03 medical and health sciences, 0302 clinical medicine, Physiology (medical), Internal medicine, Animals, Humans, Medicine, Myocytes, Cardiac, Myocardial infarction, Rats, Wistar, Cells, Cultured, 030304 developmental biology, 0303 health sciences, Secretory Pathway, Ventricular Remodeling, business.industry, Subtilisin, medicine.disease, Proprotein convertase, Mice, Inbred C57BL, Disease Models, Animal, medicine.anatomical_structure, Animals, Newborn, Cardiology, Kexin, Tissue necrosis, Blood supply, Proprotein Convertase 9, Cardiology and Cardiovascular Medicine, business, Signal Transduction, Artery |
| الوصف: | Background: Acute occlusion of a coronary artery results in swift tissue necrosis. Bordering areas of the infarcted myocardium can also experience impaired blood supply and reduced oxygen delivery, leading to altered metabolic and mechanical processes. Although transcriptional changes in hypoxic cardiomyocytes are well studied, little is known about the proteins that are actively secreted from these cells. Methods: We established a novel secretome analysis of cardiomyocytes by combining stable isotope labeling and click chemistry with subsequent mass spectrometry analysis. Further functional validation experiments included ELISA measurement of human samples, murine left anterior descending coronary artery ligation, and adeno-associated virus 9–mediated in vivo overexpression in mice. Results: The presented approach is feasible for analysis of the secretome of primary cardiomyocytes without serum starvation. A total of 1026 proteins were identified to be secreted within 24 hours, indicating a 5-fold increase in detection compared with former approaches. Among them, a variety of proteins have not yet been explored in the context of cardiovascular pathologies. One of the secreted factors most strongly upregulated upon hypoxia was PCSK6 (proprotein convertase subtilisin/kexin type 6). Validation experiments revealed an increase of PCSK6 on mRNA and protein level in hypoxic cardiomyocytes. PCSK6 expression was elevated in hearts of mice after 3 days of ligation of the left anterior descending artery, a finding confirmed by immunohistochemistry. ELISA measurements in human serum also indicate distinct kinetics for PCSK6 in patients with acute myocardial infarction, with a peak on postinfarction day 3. Transfer of PCSK6-depleted cardiomyocyte secretome resulted in decreased expression of collagen I and III in fibroblasts compared with control treated cells, and small interfering RNA–mediated knockdown of PCSK6 in cardiomyocytes impacted transforming growth factor-β activation and SMAD3 (mothers against decapentaplegic homolog 3) translocation in fibroblasts. An adeno-associated virus 9–mediated, cardiomyocyte-specific overexpression of PCSK6 in mice resulted in increased collagen expression and cardiac fibrosis, as well as decreased left ventricular function, after myocardial infarction. Conclusions: A novel mass spectrometry–based approach allows investigation of the secretome of primary cardiomyocytes. Analysis of hypoxia-induced secretion led to the identification of PCSK6 as being crucially involved in cardiac remodeling after acute myocardial infarction. |
| تدمد: | 1524-4539 0009-7322 |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::a8a54680ab13c25a0eecffd0f99832a9Test https://doi.org/10.1161/circulationaha.119.044914Test |
| حقوق: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....a8a54680ab13c25a0eecffd0f99832a9 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 15244539 00097322 |
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