المؤلفون: Harsha V. Renikunta, Katina Lazarow, Yiqi Gong, Praphulla Chandra Shukla, Vanasa Nageswaran, Hector Giral, Adelheid Kratzer, Lennart Opitz, Felix B. Engel, Arash Haghikia, Sarah Costantino, Francesco Paneni, Jens Peter von Kries, Katrin Streckfuss-Bömeke, Ulf Landmesser, Philipp Jakob

المصدر: iScience, Vol 26, Iss 5, Pp 106593- (2023)

مصطلحات موضوعية: Cardiovascular medicine, Genetics, Molecular biology, Science

الوصف: Summary: Ischemic cardiomyopathy, driven by loss of cardiomyocytes and inadequate proliferative response, persists to be a major global health problem. Using a functional high-throughput screening, we assessed differential proliferative potential of 2019 miRNAs after transient hypoxia by transfecting both miR-inhibitor and miR-mimic libraries in human iPSC-CM. Whereas miR-inhibitors failed to enhance EdU uptake, overexpression of 28 miRNAs substantially induced proliferative activity in hiPSC-CM, with an overrepresentation of miRNAs belonging to the primate-specific C19MC-cluster. Two of these miRNAs, miR-515-3p and miR-519e-3p, increased markers of early and late mitosis, indicative of cell division, and substantially alter signaling pathways relevant for cardiomyocyte proliferation in hiPSC-CM.

وصف الملف: electronic resource

العلاقة: http://www.sciencedirect.com/science/article/pii/S2589004223006703Test; https://doaj.org/toc/2589-0042Test

الوصول الحر: https://doaj.org/article/0397c180ca2842079b4aa0e59e2381c0Test

المؤلفون: Paul-Lennard Mendez, Leon Obendorf, Jerome Jatzlau, Wiktor Burdzinski, Maria Reichenbach, Vanasa Nageswaran, Arash Haghikia, Verena Stangl, Christian Hiepen, Petra Knaus

المصدر: BMC Biology, Vol 20, Iss 1, Pp 1-18 (2022)

مصطلحات موضوعية: Fluid shear stress, Endothelial cell, EndoMT, BMP, Endoglin, Caveolin, Biology (General), QH301-705.5

الوصف: Abstract Background Fluid shear stress enhances endothelial SMAD1/5 signaling via the BMP9-bound ALK1 receptor complex supported by the co-receptor Endoglin. While moderate SMAD1/5 activation is required to maintain endothelial quiescence, excessive SMAD1/5 signaling promotes endothelial dysfunction. Increased BMP signaling participates in endothelial-to-mesenchymal transition and inflammation culminating in vascular diseases such as atherosclerosis. While the function of Endoglin has so far been described under picomolar concentrations of BMP9 and short-term shear application, we investigated Endoglin under physiological BMP9 and long-term pathophysiological shear conditions. Results We report here that knock-down of Endoglin leads to exacerbated SMAD1/5 phosphorylation and atheroprone gene expression profile in HUVECs sheared for 24 h. Making use of the ligand-trap ALK1-Fc, we furthermore show that this increase is dependent on BMP9/10. Mechanistically, we reveal that long-term exposure of ECs to low laminar shear stress leads to enhanced Endoglin expression and endocytosis of Endoglin in Caveolin-1-positive early endosomes. In these endosomes, we could localize the ALK1-Endoglin complex, labeled BMP9 as well as SMAD1, highlighting Caveolin-1 vesicles as a SMAD signaling compartment in cells exposed to low atheroprone laminar shear stress. Conclusions We identified Endoglin to be essential in preventing excessive activation of SMAD1/5 under physiological flow conditions and Caveolin-1-positive early endosomes as a new flow-regulated signaling compartment for BMP9-ALK1-Endoglin signaling axis in atheroprone flow conditions.

وصف الملف: electronic resource

العلاقة: https://doaj.org/toc/1741-7007Test

الوصول الحر: https://doaj.org/article/f21e33dc96904caabcbae15c6de679caTest

المؤلفون: Martina Gast, Vanasa Nageswaran, Andreas W. Kuss, Ana Tzvetkova, Xiaomin Wang, Liliana H. Mochmann, Pegah Ramezani Rad, Stefan Weiss, Stefan Simm, Tanja Zeller, Henry Voelzke, Wolfgang Hoffmann, Uwe Völker, Stefan B. Felix, Marcus Dörr, Antje Beling, Carsten Skurk, David-Manuel Leistner, Bernhard H. Rauch, Tetsuro Hirose, Bettina Heidecker, Karin Klingel, Shinichi Nakagawa, Wolfram C. Poller, Filip K. Swirski, Arash Haghikia, Wolfgang Poller

المصدر: Cells, Vol 11, Iss 24, p 3970 (2022)

مصطلحات موضوعية: immunology, innate immunity, immunogenetics, noncoding genome, tRNA biology, evolutionary genetics, Cytology, QH573-671

الوصف: The evolutionary conserved NEAT1-MALAT1 gene cluster generates large noncoding transcripts remaining nuclear, while tRNA-like transcripts (mascRNA, menRNA) enzymatically generated from these precursors translocate to the cytosol. Whereas functions have been assigned to the nuclear transcripts, data on biological functions of the small cytosolic transcripts are sparse. We previously found NEAT1−/− and MALAT1−/− mice to display massive atherosclerosis and vascular inflammation. Here, employing selective targeted disruption of menRNA or mascRNA, we investigate the tRNA-like molecules as critical components of innate immunity. CRISPR-generated human ΔmascRNA and ΔmenRNA monocytes/macrophages display defective innate immune sensing, loss of cytokine control, imbalance of growth/angiogenic factor expression impacting upon angiogenesis, and altered cell–cell interaction systems. Antiviral response, foam cell formation/oxLDL uptake, and M1/M2 polarization are defective in ΔmascRNA/ΔmenRNA macrophages, defining first biological functions of menRNA and describing new functions of mascRNA. menRNA and mascRNA represent novel components of innate immunity arising from the noncoding genome. They appear as prototypes of a new class of noncoding RNAs distinct from others (miRNAs, siRNAs) by biosynthetic pathway and intracellular kinetics. Their NEAT1-MALAT1 region of origin appears as archetype of a functionally highly integrated RNA processing system.

وصف الملف: electronic resource

العلاقة: https://www.mdpi.com/2073-4409/11/24/3970Test; https://doaj.org/toc/2073-4409Test

الوصول الحر: https://doaj.org/article/ec8ff563749948efa86a683c898e23a4Test

المؤلفون: Martina Gast, Vanasa Nageswaran, Andreas Kuss, Ana Tzvetkova, Xiaomin Wang, Liliana Mochmann, Pegah Ramezani-Rad, Stefan Weiss, Stefan Simm, Tanja Zeller, Henry Voelzke, Wolfgang Hoffmann, Uwe Völker, Stefan Felix, Marcus Dörr, Antje Beling, Carsten Skurk, David-Manuel Leistner, Bernhard Rauch, Tetsuro Hirose, Bettina Heidecker, Karin Klingel, Shinichi Nakagawa, Wolfram Poller, Filip Swirski, Arash Haghikia, Wolfgang Poller

الوصف: The evolutionary conserved NEAT1-MALAT1 gene cluster generates large noncoding transcripts remaining nuclear, while tRNA-like transcripts (mascRNA, menRNA) enzymatically generated from these precursors translocate to the cytosol. NEAT1-/- and MALAT1-/- mice display massive atherosclerosis and vascular inflammation. Here, we identify the tRNA-like molecules as critical components of innate immunity. They appear as prototypes of a new class of noncoding RNAs distinct from others (miRNAs, siRNAs) by biosynthetic pathway and intracellular kinetics. CRISPR-generated human ΔmascRNA and ΔmenRNA monocytes/macrophages display defective innate immune sensing, loss of cytokine control, imbalance of growth/angiogenic factor expression impacting upon angiogenesis, and altered cell-cell interaction systems. Antiviral response, foam cell formation/oxLDL uptake, and M1/M2 polarization are defective in ΔmascRNA/ΔmenRNA macrophages, defining the tRNA-like molecules’ first described biological functions. menRNA and mascRNA represent novel components of innate immunity arising from the noncoding genome. Their NEAT1-MALAT1 region of origin appears as archetype of a functionally highly integrated RNA processing system.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::56a7a42e75f290709b2a3e0362feecb5Test
https://doi.org/10.21203/rs.3.rs-1540419/v2Test

المؤلفون: Chinmay Kumar Dwibedi, Ahmed N. Hegazy, Johannes Kaisler, David M. Leistner, Annette Aigner, Aiden Haghikia, Stanley L. Hazen, Yvonne Döring, Valentina Tremaroli, Arash Haghikia, Nicolle Kränkel, David Schmidt, Markus M. Heimesaat, Paul Schumann, Christopher M. Strauch, Andrzej Jasina, Stefan Bereswill, Philipp Heinze, Ute Seeland, Dominik N. Müller, Geraldine Rauch, Ina Nemet, Fredrik Bäckhed, Johann Roessler, Uta Ceglarek, Oliver Soehnlein, Ralf Gold, Emiel P. C. van der Vorst, Ulf Landmesser, Friederike Zimmermann, Roodline Cineus, Vanasa Nageswaran

المساهمون: Pathologie, RS: Carim - B07 The vulnerable plaque: makers and markers

المصدر: European Heart Journal, 43(6). Oxford University Press

مصطلحات موضوعية: Apolipoprotein E, medicine.medical_specialty, DIETARY FIBER, Metabolite, PROTEIN, Placebo, Lesion, Mice, chemistry.chemical_compound, Apolipoproteins E, Immune system, Internal medicine, PHOSPHATIDYLCHOLINE, medicine, Animals, Humans, Mice, Knockout, Gut microbiome, business.industry, Cholesterol, CARDIOVASCULAR RISK, MORTALITY, Interleukin, Cholesterol, LDL, Atherosclerosis, Propionic acid, MICROBIOTA, Mice, Inbred C57BL, Endocrinology, CHAIN FATTY-ACIDS, Intestinal Absorption, chemistry, Cardiovascular and Metabolic Diseases, CELLS, Intestinal cholesterol absorption, lipids (amino acids, peptides, and proteins), Propionates, medicine.symptom, Cardiology and Cardiovascular Medicine, business, 610 Medizin und Gesundheit, TRIMETHYLAMINE-N-OXIDE

الوصف: Aims Atherosclerotic cardiovascular disease (ACVD) is a major cause of mortality and morbidity worldwide, and increased low-density lipoproteins (LDLs) play a critical role in development and progression of atherosclerosis. Here, we examined for the first time gut immunomodulatory effects of the microbiota-derived metabolite propionic acid (PA) on intestinal cholesterol metabolism. Methods and results Using both human and animal model studies, we demonstrate that treatment with PA reduces blood total and LDL cholesterol levels. In apolipoprotein E−/− (Apoe −/−) mice fed a high-fat diet (HFD), PA reduced intestinal cholesterol absorption and aortic atherosclerotic lesion area. Further, PA increased regulatory T-cell numbers and interleukin (IL)-10 levels in the intestinal microenvironment, which in turn suppressed the expression of Niemann-Pick C1-like 1 (Npc1l1), a major intestinal cholesterol transporter. Blockade of IL-10 receptor signalling attenuated the PA-related reduction in total and LDL cholesterol and augmented atherosclerotic lesion severity in the HFD-fed Apoe −/− mice. To translate these preclinical findings to humans, we conducted a randomized, double-blinded, placebo-controlled human study (clinical trial no. NCT03590496). Oral supplementation with 500 mg of PA twice daily over the course of 8 weeks significantly reduced LDL [−15.9 mg/dL (−8.1%) vs. −1.6 mg/dL (−0.5%), P = 0.016], total [−19.6 mg/dL (−7.3%) vs. −5.3 mg/dL (−1.7%), P = 0.014] and non-high-density lipoprotein cholesterol levels [PA vs. placebo: −18.9 mg/dL (−9.1%) vs. −0.6 mg/dL (−0.5%), P = 0.002] in subjects with elevated baseline LDL cholesterol levels. Conclusion Our findings reveal a novel immune-mediated pathway linking the gut microbiota-derived metabolite PA with intestinal Npc1l1 expression and cholesterol homeostasis. The results highlight the gut immune system as a potential therapeutic target to control dyslipidaemia that may introduce a new avenue for prevention of ACVDs.

وصف الملف: application/pdf

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::ce082ae98013c4c937e3c28c224c445aTest
https://doi.org/10.1093/eurheartj/ehab644Test

المؤلفون: Fabrizio Montecucco, Thomas F. Lüscher, Luca Liberale, Daniel S. Gaul, Giacomo Giacalone, Daria Vdovenko, Sarah Costantino, Julien Weber, Jürgen Pahla, Urs Eriksson, Maria Sessa, Giovanni G. Camici, Vanasa Nageswaran, Vera Fehr, Gerd A. Kullak-Ublick, Alexander Akhmedov, Aurora Semerano, Frank Ruschitzka, Christian M. Matter, Jürg H. Beer, Nicole R. Bonetti, Francesco Paneni

المساهمون: University of Zurich, Camici, Giovanni G

المصدر: European Heart Journal. 41:1575-1587

مصطلحات موضوعية: Cell death, medicine.medical_specialty, Endothelium, Ischemia, 610 Medicine & health, 030204 cardiovascular system & hematology, Blood–brain barrier, Neuroprotection, 2705 Cardiology and Cardiovascular Medicine, Brain Ischemia, 11459 Center for Molecular Cardiology, Brain ischemia, Mice, 03 medical and health sciences, 0302 clinical medicine, Internal medicine, Sirtuin 6, SIRT6, Animals, Humans, Sirtuins, Medicine, Middle cerebral artery occlusion, Stroke, 030304 developmental biology, 0303 health sciences, business.industry, Endothelial Cells, Infarction, Middle Cerebral Artery, Human brain, Ischaemia/reperfusion, Hypoxia (medical), medicine.disease, Mice, Inbred C57BL, medicine.anatomical_structure, Endocrinology, Blood-Brain Barrier, 10199 Clinic for Clinical Pharmacology and Toxicology, 10209 Clinic for Cardiology, medicine.symptom, Cardiology and Cardiovascular Medicine, business

الوصف: Aims Aging is an established risk factor for stroke; genes regulating longevity are implicated in the pathogenesis of ischaemic stroke where to date, therapeutic options remain limited. The blood–brain barrier (BBB) is crucially involved in ischaemia/reperfusion (I/R) brain injury thus representing an attractive target for developing novel therapeutic agents. Given the role of endothelial cells in the BBB, we hypothesized that the endothelial-specific expression of the recently described longevity gene SIRT6 may exhibit protective properties in stroke. Methods and results SIRT6 endothelial expression was reduced following stroke. Endothelial-specific Sirt6 knockout (eSirt6−/−) mice, as well as animals in which Sirt6 overexpression was post-ischaemically induced, underwent transient middle cerebral artery occlusion (tMCAO). eSirt6−/− animals displayed increased infarct volumes, mortality, and neurological deficit after tMCAO, as compared to control littermates. Conversely, post-ischaemic Sirt6 overexpression decreased infarct size and neurological deficit. Analysis of ischaemic brain sections revealed increased BBB damage and endothelial expression of cleaved caspase-3 in eSIRT6−/− mice as compared to controls. In primary human brain microvascular endothelial cells (HBMVECs), hypoxia/reoxygenation (H/R) reduced SIRT6 expression and SIRT6 silencing impaired the barrier function (transendothelial resistance) similar to what was observed in mice exposed to I/R. Further, SIRT6-silenced HBMVECs exposed to H/R showed reduced viability, increased cleaved caspase-3 expression and reduced activation of the survival pathway Akt. In ischaemic stroke patients, SIRT6 expression was higher in those with short-term neurological improvement as assessed by NIHSS scale and correlated with stroke outcome. Conclusion Endothelial SIRT6 exerts a protective role in ischaemic stroke by blunting I/R-mediated BBB damage and thus, it may represent an interesting novel therapeutic target to be explored in future clinical investigation.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::9bdf56c4e49b9522a247eb1254f9b1e3Test
https://doi.org/10.1093/eurheartj/ehz712Test

المؤلفون: Antje Beling, Martina Gast, A Tzvetkova, L Mochmann, Bernhard H. Rauch, Arash Haghikia, Andreas W. Kuss, Karin Klingel, Xiaomin Wang, Ulf Landmesser, Vanasa Nageswaran, Stefan Weiss, C Poller, Tanja Zeller, Shinichi Nakagawa

المصدر: European Heart Journal. 42

مصطلحات موضوعية: Class (set theory), MALAT1, Innate immune system, business.industry, Angiogenesis, Transfer RNA, Medicine, Computational biology, Cardiology and Cardiovascular Medicine, business

الوصف: Background The evolutionary conserved NEAT1-MALAT1 gene cluster encounters high interest in cardiovascular medicine and oncology. The cluster generates large primary transcripts which remain nuclear, whereas novel tRNA-like transcripts (mascRNA, menRNA) enzymatically generated from these precursors translocate to the cytosol. We previously found that NEAT1 and MALAT1 deficient mice display accelerated atherosclerosis and vascular inflammation due to immune dysfunctions. Methods While the previously investigated mice were deficient in the entire NEAT1 or MALAT1 locus, here we aimed to selectively disrupt only tRNA-like transcripts “menRNA” arising from NEAT1, or “mascRNA” arising from MALAT1. To none of these a biological function has been assigned so far. Both lncRNAs give rise to transcripts of vastly different size (NEAT1: 23kb MENb, 3.7kb MENe, 59nt “menRNA”; MALAT1: 8.3 kb primary, 59nt “mascRNA”), and traditional knockout methods are unable to selectively inactivate one of the small transcripts only. Through CRISPR/Cas9 editing we therefore developed human monocyte-macrophage cell lines with short deletions in the respective tRNA-encoding sequences to disrupt normal menRNA or mascRNA formation, respectively. These editing procedures do not affect transcription of the respective lncRNA parent transcripts, and also not disturb regular formation of the triple-helix structures at their 3'-ends which support stabilization of the respective lncRNAs (Fig. 1). Results We found the tRNA-like transcripts menRNA and mascRNA critically influence innate immunity and angiogenesis. In addition to common anomalies resulting from their selective CRISPR-Cas9 mediated deletion (Fig. 1), there are specific disturbances associated with either Δmasc or Δmen cells (Fig. 2). Both ΔmascRNA and ΔmenRNA human monocytes show profoundly altered ribosomal RNA/protein and tRNA-modifying enzyme expression, display anomalous growth/ angiogenetic factor expression, fundamentally change angiogenetic patterns in co-cultures with human endothelial cells, and have gravely disturbed innate immune responses (LPS, DNA and RNA viruses) (Fig. 1). CRISPR-engineered ΔmenRNA cells share remakable similarities with human post-MI PBMCs, suggesting the NEAT1-menRNA system may significantly contribute to post-MI residual inflammatory risk despite optimal standard therapy (Fig. 2). Conclusions Beyond prior work in knockout mice documenting immune function of the NEAT1-MALAT1 cluster, the current study identifies menRNA and mascRNA as important novel components of human innate immunity with relevance for angiogenetic processes. These data provide a second mechanistic link for the apparent relevance of the NEAT1-MALAT1 gene cluster in cardiovascular and malignant diseases. As prototypes of a novel class of small noncoding RNAs (distinct from miRNAs and siRNAs) they may constitute cytosolic therapeutic targets. Funding Acknowledgement Type of funding sources: Other. Main funding source(s): DZHK Shared Expertise Project/B19-006_SE/FKZ 81X2100257/Transcriptome analysis of circulating immune cells to improve the assessment of prognosis and the response to novel anti-inflammatory treatments after myocardial infarction Figure 1. Common anomaliesFigure 2. Specific anomalies

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_________::31f2f0df7d99f339ba1e355ab6d5898eTest
https://doi.org/10.1093/eurheartj/ehab724.3357Test

المؤلفون: Frank Ruschitzka, Kimon Stamatelopoulos, Luca Liberale, Giovanni G. Camici, Melroy X. Miranda, Nicole R. Bonetti, Vanasa Nageswaran, Konstantinos Stellos, Francesco Paneni, Jürg H. Beer, Alexander Akhmedov, Nikolaos I. Vlachogiannis, Fabrizio Montecucco, Thomas F. Lüscher, Sarah Costantino, Yustina M. Puspitasari, Lena Schwarz

المساهمون: University of Zurich

مصطلحات موضوعية: Male, Physiology, medicine.medical_treatment, PAI-1, 030204 cardiovascular system & hematology, Pharmacology, arterial thrombosis, AMP-Activated Protein Kinases, chemistry.chemical_compound, 0302 clinical medicine, cardiovascular disease, Sirtuin 5, Sirtuins, Platelet, Phosphorylation, Extracellular Signal-Regulated MAP Kinases, Cells, Cultured, Mice, Knockout, 0303 health sciences, biology, Fibrinolysis, Middle Aged, Thrombosis, 3. Good health, medicine.anatomical_structure, Plasminogen activator inhibitor-1, Sirtuin, 10209 Clinic for Cardiology, Female, Cardiology and Cardiovascular Medicine, SIRT5, tissue factor, Adult, Endothelium, 610 Medicine & health, 03 medical and health sciences, Physiology (medical), Plasminogen Activator Inhibitor 1, medicine, Animals, Humans, Carotid Artery Thrombosis, Thrombus, Acute Coronary Syndrome, 030304 developmental biology, Aged, business.industry, Endothelial Cells, medicine.disease, Mice, Inbred C57BL, Disease Models, Animal, chemistry, Case-Control Studies, biology.protein, business, Carotid Artery Injuries, Plasminogen activator

الوصف: AIMS Arterial thrombosis as a result of plaque rupture or erosion is a key event in acute cardiovascular events. Sirtuin 5 (SIRT5) belongs to the lifespan-regulating sirtuin superfamily and has been implicated in acute ischemic stroke and cardiac hypertrophy. This project aims at investigating the role of SIRT5 in arterial thrombus formation. METHODS AND RESULTS Sirt5 transgenic (Sirt5Tg/0) as well as knock-out (Sirt5-/-) mice underwent photochemically-induced carotid endothelial injury to trigger arterial thrombosis. Primary human aortic endothelial cells (HAECs) treated with SIRT5 silencing-RNA (si-SIRT5) as well as peripheral blood mononuclear cells (PBMCs) from acute coronary syndrome (ACS) patients and non-ACS controls (case-control study, total n = 171) were used to increase the translational relevance of our data. Compared to WT controls, Sirt5Tg/0 mice displayed accelerated arterial thrombus formation following endothelial-specific damage. Conversely, in Sirt5-/-mice arterial thrombosis was blunted. Platelet function was unaltered, as assessed by ex vivo collagen-induced aggregometry. Similarly, activation of the coagulation cascade as assessed by vascular and plasma tissue factor (TF) and TF pathway inhibitor (TFPI) expression was unaltered. Increased thrombus embolization episodes and circulating D-dimer levels suggested augmented activation of the fibrinolytic system in Sirt5-/- mice. Accordingly, Sirt5-/- mice showed reduced plasma and vascular expression of the fibrinolysis inhibitor plasminogen activator inhibitor (PAI)-1. In HAECs, SIRT5-silencing inhibited PAI-1 gene and protein expression in response to TNF-α. This effect was mediated by increased AMPK activation and reduced phosphorylation of the MAP kinase ERK 1/2, but not JNK and p38 as shown both in vivo and in vitro. Lastly, both PAI-1 and SIRT5 gene expression are increased in ACS patients compared to non-ACS controls after adjustment for cardiovascular risk factors, while PAI-1 expression increased across tertiles of SIRT5. CONCLUSIONS SIRT5 promotes arterial thrombosis by modulating fibrinolysis through endothelial PAI-1 expression. Hence, SIRT5 may be an interesting therapeutic target in the context of atherothrombotic events. TRANSLATIONAL PERSPECTIVES This study illustrates a novel role for Sirtuin 5 in arterial thrombosis by regulating fibrinolysis through plasminogen activator inhibitor 1 (PAI-1). These results shed new light onto the pathophysiology of arterial thrombus formation which underlies most of the acute atherosclerotic complications. Also, they further affirm the intrinsic relationship between lifespan regulating genes, vascular dysfunction and age-related cardiovascular disease, thus indicating these genes as potential targets for cardiovascular prevention and therapy. Further studies will be needed to assess the predictive ability of SIRT5 in patients with acute cardiovascular or cerebrovascular events. Also, the design of specific SIRT5 inhibitors will allow trials aiming at investigating the efficacy of SIRT5 blockage in the clinical setting.

وصف الملف: cvaa268.pdf - application/pdf

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::4ebda8ee2d276530ca3818d24aab3c8bTest

المؤلفون: Patricia Wüst, Fabrizio Montecucco, Thomas F. Lüscher, Jürg H. Beer, Candela Diaz-Cañestro, Luca Liberale, Johannes Bohacek, Giovanni G. Camici, Nicole R. Bonetti, Vanasa Nageswaran

المساهمون: University of Zurich, Camici, Giovanni G

مصطلحات موضوعية: 1303 Biochemistry, Platelet Aggregation, Clinical Biochemistry, arterial thrombosis, 030204 cardiovascular system & hematology, 1308 Clinical Biochemistry, Biochemistry, 11459 Center for Molecular Cardiology, Mice, Phosphatidylinositol 3-Kinases, 0302 clinical medicine, Apold1, platelets, tissue factor, Platelet, 030212 general & internal medicine, Extracellular Signal-Regulated MAP Kinases, Mice, Knockout, Laser Coagulation, Kinase, Chemistry, General Medicine, Photochemical Processes, 10209 Clinic for Cardiology, Signal Transduction, Blood Platelets, medicine.medical_specialty, Platelet Function Tests, MAP Kinase Signaling System, Context (language use), 610 Medicine & health, Real-Time Polymerase Chain Reaction, Collagen Type I, Immediate-Early Proteins, Thromboplastin, 03 medical and health sciences, Tissue factor, In vivo, Internal medicine, medicine, Animals, Carotid Artery Thrombosis, RNA, Messenger, Thrombus, Protein kinase B, PI3K/AKT/mTOR pathway, Fluorescent Dyes, Rose Bengal, Endothelial Cells, medicine.disease, Endocrinology, Proto-Oncogene Proteins c-akt

الوصف: BACKGROUND Endothelial cells regulate the formation of blood clots thus, genes selectively expressed in these cells could primarily determine thrombus formation. Apold1(apolipoprotein L domain containing 1) is a gene expressed by endothelial cells; whether Apold1 directly contributes to arterial thrombosis has not yet been investigated. Here, we assessed the effect of Apold1 deletion on arterial thrombus formation using an in vivo model of carotid thrombosis induced by photochemical injury. MATERIAL AND METHODS Apold1 knockout (Apold1-/- ) mice and wild type (WT) littermates underwent carotid thrombosis induced by photochemical injury and time to occlusion was recorded. Tissue factor (TF) activity as well as activation of mitogen-activated protein kinases (MAPKs) and phosphatidyl-inositol-3 kinase (PI3K)/Akt pathways were analyzed by colorimetric assay and western blotting in both Apold1-/- and WT mice. Finally, platelet reactivity was assessed using light transmission aggregometry. RESULTS After photochemical injury, Apold1$^{-/-}$ mice exhibited shorter time to occlusion as compared to WT mice. Moreover, TF activity was increased in carotid arteries of Apold1$^{-/-}$ when compared to WT mice. Underlying mechanistic markers such as TF mRNA and MAPKs activation were unaffected in Apold1$^{-/-}$ mice. In contrast, phosphorylation of Akt was reduced in Apold1$^{-/-}$ as compared to WT mice. Additionally, Apold1$^{-/-}$ mice displayed increased platelet reactivity to stimulation with collagen compared to WT animals. CONCLUSIONS Deficiency of Apold1 results in a prothrombotic phenotype, accompanied by increased vascular TF activity,decreased PI3K/Akt activation and increased platelet reactivity. These findings suggest Apold1 as an interesting new therapeutic target in the context of arterial thrombosis.

وصف الملف: Revised_Manuscript_Apold1_final.pdf - application/pdf

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::e77594541e2c8e3787f24f05f6d298aaTest
https://doi.org/10.5167/uzh-180309Test

المؤلفون: J H Beer, Christian M. Matter, Nicole R. Bonetti, Thomas F Luescher, Fabrizio Montecucco, Vanasa Nageswaran, G G Camici, Luca Liberale, Francesco Paneni, S Costantino, Alexander Akhmedov, Jürgen Pahla

المصدر: European Heart Journal. 40

مصطلحات موضوعية: SIRT6, medicine.medical_specialty, medicine.anatomical_structure, business.industry, Internal medicine, Ischemia, medicine, Cardiology, Cardiology and Cardiovascular Medicine, medicine.disease, business, Blood–brain barrier, Reperfusion injury

الوصف: Introduction Stroke is a major cause of mortality and morbidity worldwide. Yet, therapeutic strategies are limited to the early reperfusion which can, on the other hand, worsen the brain damage trough ischemia/reperfusion (I/R) injury. Post-stroke blood-brain barrier (BBB) impairment is associated with worsened outcome. Aging is a major risk factor for stroke and genes regulating lifespan also contribute to the determination of cerebral damage during I/R injury. Purpose Given the pivotal role of endothelial cells in BBB, we hypothesized that the endothelial-specific expression of the longevity gene SIRT6 may protect the BBB from ischemia/reperfusion damage thus having a beneficial role on stroke outcome. Methods Endothelial-specific SIRT6 knockout (eSIRT6−/−) mice and control littermates (CTRL) underwent transient middle cerebral artery occlusion (tMCAO) for 45 min followed by 48 hours of reperfusion. Immunohistochemistry (IHC) was used to investigate BBB permeability by IgG extravasation and molecular mechanisms. Primary human brain microvascular endothelial cells (HBMVECs) transfected with either SIRT6 (siSIRT6) or scrambled (siSCR) small interfering RNA were subjected to hypoxia/reoxygenation (H/R). An in vitro BBB model consisting of a monolayer of siRNA-treated HBMVECs was established and barrier function was assessed by 48 h-lasting transendothelial electrical resistance measurement. SIRT6 expression in monocytes from stroke patients was correlated with the short-term neurological outcome [ΔNIHSS% = (NIHSS discharge-NIHSS admission)/ NIHSS admission*100]. Results eSIRT6−/− displayed higher infarct volumes and lower survival rate compared to WT mice 48 h after tMCAO. The increased infarct volume was functionally relevant as eSIRT6−/− also showed worse post-stroke neurological impairment. Analysis of brain sections revealed increased BBB damage and increased endothelial expression of cleaved caspase-3 in eSIRT6−/− as compared to control littermates. In vitro, H/R reduced SIRT6 expression in HBMVECs. Mirroring the animal results, SIRT6 silencing impaired the barrier function of HBMVECs 48 h after exposure to H/R. In line with this, SIRT6-silenced HBMVECs showed reduced viability, increased cleaved caspase-3 expression and reduced activation of the anti-apoptotic survival pathway Akt as compared to control cells after H/R. The direct interaction between SIRT6 and Akt was confirmed by co-immunoprecipitation. In ischemic stroke patients, SIRT6 expression was higher in those with short-term neurological improvement (ΔNIHSS% >0) and negatively correlated with ΔNIHSS%. Conclusion Endothelial SIRT6 exerts a beneficial role in ischemic stroke by blunting I/R-mediated BBB damage. Specifically, SIRT6 reduces endothelial I/R-induced apoptotic death through activation of the protective Akt pathway. The longevity gene SIRT6 may represent a novel therapeutic target for the treatment of ischemic stroke. Acknowledgement/Funding Swiss National Science Foundation

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_________::4e416f839924cc34b8f960653e493480Test
https://doi.org/10.1093/eurheartj/ehz748.0132Test