RESUMEN
BACKGROUND: Recognition of the importance of conventional lipid measures and the advent of novel lipid-lowering medications have prompted the need for more comprehensive lipid panels to guide use of emerging treatments for the prevention of coronary heart disease (CHD). This report assessed the relevance of 13 apolipoproteins measured using a single mass-spectrometry assay for risk of CHD in the PROCARDIS case-control study of CHD (941 cases/975 controls). METHODS: The associations of apolipoproteins with CHD were assessed after adjustment for established risk factors and correction for statin use. Apolipoproteins were grouped into 4 lipid-related classes [lipoprotein(a), low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, and triglycerides] and their associations with CHD were adjusted for established CHD risk factors and conventional lipids. Analyses of these apolipoproteins in a subset of the ASCOT trial (Anglo-Scandinavian Cardiac Outcomes Trial) were used to assess their within-person variability and to estimate a correction for statin use. The findings in the PROCARDIS study were compared with those for incident cardiovascular disease in the Bruneck prospective study (n=688), including new measurements of Apo(a). RESULTS: Triglyceride-carrying apolipoproteins (ApoC1, ApoC3, and ApoE) were most strongly associated with the risk of CHD (2- to 3-fold higher odds ratios for top versus bottom quintile) independent of conventional lipid measures. Likewise, ApoB was independently associated with a 2-fold higher odds ratios of CHD. Lipoprotein(a) was measured using peptides from the Apo(a)-kringle repeat and Apo(a)-constant regions, but neither of these associations differed from the association with conventionally measured lipoprotein(a). Among HDL-related apolipoproteins, ApoA4 and ApoM were inversely related to CHD, independent of conventional lipid measures. The disease associations with all apolipoproteins were directionally consistent in the PROCARDIS and Bruneck studies, with the exception of ApoM. CONCLUSIONS: Apolipoproteins were associated with CHD independent of conventional risk factors and lipids, suggesting apolipoproteins could help to identify patients with residual lipid-related risk and guide personalized approaches to CHD risk reduction.
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Enfermedad Coronaria , Inhibidores de Hidroximetilglutaril-CoA Reductasas , Humanos , Estudios Prospectivos , Inhibidores de Hidroximetilglutaril-CoA Reductasas/uso terapéutico , Estudios de Casos y Controles , Proteómica , Apolipoproteínas , Factores de Riesgo , Enfermedad Coronaria/epidemiología , Enfermedad Coronaria/etiología , Triglicéridos , HDL-Colesterol , Lipoproteína(a) , Apolipoproteínas B/uso terapéutico , Apolipoproteína A-IRESUMEN
BACKGROUND: Using proteomics, we aimed to reveal molecular types of human atherosclerotic lesions and study their associations with histology, imaging, and cardiovascular outcomes. METHODS: Two hundred nineteen carotid endarterectomy samples were procured from 120 patients. A sequential protein extraction protocol was employed in conjunction with multiplexed, discovery proteomics. To focus on extracellular proteins, parallel reaction monitoring was employed for targeted proteomics. Proteomic signatures were integrated with bulk, single-cell, and spatial RNA-sequencing data, and validated in 200 patients from the Athero-Express Biobank study. RESULTS: This extensive proteomics analysis identified plaque inflammation and calcification signatures, which were inversely correlated and validated using targeted proteomics. The inflammation signature was characterized by the presence of neutrophil-derived proteins, such as S100A8/9 (calprotectin) and myeloperoxidase, whereas the calcification signature included fetuin-A, osteopontin, and gamma-carboxylated proteins. The proteomics data also revealed sex differences in atherosclerosis, with large-aggregating proteoglycans versican and aggrecan being more abundant in females and exhibiting an inverse correlation with estradiol levels. The integration of RNA-sequencing data attributed the inflammation signature predominantly to neutrophils and macrophages, and the calcification and sex signatures to smooth muscle cells, except for certain plasma proteins that were not expressed but retained in plaques, such as fetuin-A. Dimensionality reduction and machine learning techniques were applied to identify 4 distinct plaque phenotypes based on proteomics data. A protein signature of 4 key proteins (calponin, protein C, serpin H1, and versican) predicted future cardiovascular mortality with an area under the curve of 75% and 67.5% in the discovery and validation cohort, respectively, surpassing the prognostic performance of imaging and histology. CONCLUSIONS: Plaque proteomics redefined clinically relevant patient groups with distinct outcomes, identifying subgroups of male and female patients with elevated risk of future cardiovascular events.
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Aterosclerosis , Calcinosis , Femenino , Humanos , Masculino , Proteómica , Caracteres Sexuales , Versicanos , alfa-2-Glicoproteína-HSRESUMEN
The right ventricle (RV) differs developmentally, anatomically and functionally from the left ventricle (LV). Therefore, characteristics of LV adaptation to chronic pressure overload cannot easily be extrapolated to the RV. Mitochondrial abnormalities are considered a crucial contributor in heart failure (HF), but have never been compared directly between RV and LV tissues and cardiomyocytes. To identify ventricle-specific mitochondrial molecular and functional signatures, we established rat models with two slowly developing disease stages (compensated and decompensated) in response to pulmonary artery banding (PAB) or ascending aortic banding (AOB). Genome-wide transcriptomic and proteomic analyses were used to identify differentially expressed mitochondrial genes and proteins and were accompanied by a detailed characterization of mitochondrial function and morphology. Two clearly distinguishable disease stages, which culminated in a comparable systolic impairment of the respective ventricle, were observed. Mitochondrial respiration was similarly impaired at the decompensated stage, while respiratory chain activity or mitochondrial biogenesis were more severely deteriorated in the failing LV. Bioinformatics analyses of the RNA-seq. and proteomic data sets identified specifically deregulated mitochondrial components and pathways. Although the top regulated mitochondrial genes and proteins differed between the RV and LV, the overall changes in tissue and cardiomyocyte gene expression were highly similar. In conclusion, mitochondrial dysfuntion contributes to disease progression in right and left heart failure. Ventricle-specific differences in mitochondrial gene and protein expression are mostly related to the extent of observed changes, suggesting that despite developmental, anatomical and functional differences mitochondrial adaptations to chronic pressure overload are comparable in both ventricles.
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Modelos Animales de Enfermedad , Insuficiencia Cardíaca , Mitocondrias Cardíacas , Animales , Mitocondrias Cardíacas/metabolismo , Mitocondrias Cardíacas/patología , Masculino , Insuficiencia Cardíaca/metabolismo , Insuficiencia Cardíaca/fisiopatología , Insuficiencia Cardíaca/patología , Insuficiencia Cardíaca/genética , Proteómica , Disfunción Ventricular Derecha/fisiopatología , Disfunción Ventricular Derecha/metabolismo , Disfunción Ventricular Derecha/genética , Disfunción Ventricular Derecha/patología , Función Ventricular Derecha , Miocitos Cardíacos/metabolismo , Miocitos Cardíacos/patología , Ventrículos Cardíacos/metabolismo , Ventrículos Cardíacos/fisiopatología , Ventrículos Cardíacos/patología , Ratas , Función Ventricular Izquierda , Disfunción Ventricular Izquierda/fisiopatología , Disfunción Ventricular Izquierda/metabolismo , Disfunción Ventricular Izquierda/patología , Disfunción Ventricular Izquierda/genética , Transcriptoma , Ratas Sprague-Dawley , Proteínas Mitocondriales/metabolismo , Proteínas Mitocondriales/genéticaRESUMEN
Cladding-pumped multicore erbium-doped fiber is an important element for future spatial division multiplexing (SDM) amplification. We propose an M-type erbium-doped multicore fiber to achieve high-efficiency SDM amplification. The performance of cladding-pumped erbium-doped fiber with a central refractive index depression has been investigated, and the M-type fiber has better amplification performance than conventional fibers by reducing the signal mode overlap with the doped region. The experiment results show that the M-type 4-core erbium-doped fiber has a gain improvement of 2.8â dB compared with conventional 4-core fiber. The pump conversion efficiency (PCE) has been enhanced from 4.47% to 8.01%. For a 7.0 W pump power at 976â nm, the M-type fiber exhibits an average gain of 20.0â dB and an average noise fiber of 6.8â dB at the L-band. The core-to-core gain variation is less than 1.6â dB.
RESUMEN
Bismuth-doped germanosilicate fiber (BGSF), the active media of fiber amplifiers, has attracted widespread attention. Here, we report a BGSF with a high bismuth concentration of 0.075â wt. % and achieve high-efficiency E + S-band amplification, which was prepared by the modified chemical vapor deposition (MCVD) process. The small signal absorption (SSA) and unsaturated loss (UL) of BGSF at 1310â nm are 1.32 and 0.11â dB/m, respectively. The results show a record with only 45 m BGSF was created, to the best of our knowledge, which provides a maximum gain of 39.24â dB with an NF of 6.2â dB at 1430â nm under -20â dBm input signal power.
RESUMEN
Extending the gain bandwidth of L-band optical fiber amplifier has provoked a widespread interest. To date, achieving a high-efficiency extended L-band amplification remains a challenge. Here, we report a cladding-pumped Er/Yb co-doped alumino-phosphosilicate fiber, prepared by the modified chemical vapor deposition process. We demonstrate the efficiency of alumino-phosphosilicate glass for cladding-pumped Er/Yb co-doped fiber, with a gain per unit fiber length of 0.45â dB/m at 1625â nm and a gain ripple of â¼9.4â dB. For 0.8â W pump power, the fiber exhibits a 20â dB gain bandwidth covering 1575-1625â nm and 6.9â dB noise figure at 1625â nm. Additionally, the utilization of multi-mode laser diode enables further significant power savings and cost reduction. To the best of our knowledge, Er/Yb co-doped fiber in alumino-phosphosilicate glass is first proposed, with a cladding-pumped scheme for enhancing an extended L-band performance.
RESUMEN
The extended L-band 4-core Er/Yb co-doped fiber and amplifier (MC-EYDFA) is first proposed and demonstrated, to the best of our knowledge, for space division multiplexing combined with wavelength division multiplexing application. The fiber core co-doped with Er/Yb/P is adopted for bandwidth expansion, and the long wavelength extends to 1625â nm. Numerical simulations further show that efficient amplification and higher saturation power are achieved with the 1018â nm cladding pumping. Based on the integrated 4-core fiber amplifier, an average gain of â¼22â dB covering 1575-1625â nm is experimentally obtained with a 4 W pump power and a 3 dBm total signal power, and the max core-dependent gain (CDG) variation is measured to be 1.7â dB.
RESUMEN
The 1.5-µm fiber laser is widely used in the fields of laser lidar, remote sensing, and gas monitoring because of its advantages of being eye-safe and exhibiting low atmospheric transmission loss. However, due to the â¼1-µm amplified spontaneous emission (ASE) of the Er/Yb co-doped fiber (EYDF), it is difficult to improve the laser power. Here, we simulated the effect of the Er3+ concentration and the seed power on â¼1-µm ASE, and fabricated a large mode area EYDF by the modified chemical vapor deposition process. Additionally, a piece of ytterbium-doped fiber was introduced into the master oscillator power amplifier (MOPA) configuration to absorb the generated â¼1-µm ASE simultaneously. Experimental results show that an output power of 345 W with a slope efficiency of 43% at 1535â nm is obtained in an all-fiber configuration, profiting from effective suppression of â¼ 1-µm ASE. To the best of our knowledge, this is the highest output power available with an Er/Yb co-doped fiber from an all-fiber MOPA configuration.
RESUMEN
[Figure: see text].
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Fibrilación Atrial/metabolismo , Miocardio/metabolismo , Proteínas Serina-Treonina Quinasas/metabolismo , Anciano , Animales , Fibrilación Atrial/tratamiento farmacológico , Fibrilación Atrial/genética , Fibrilación Atrial/patología , Células Cultivadas , Femenino , Fibrosis , Humanos , Masculino , Mesalamina/farmacología , Mesalamina/uso terapéutico , Ratones , Ratones Endogámicos C57BL , Persona de Mediana Edad , Proteína Quinasa 1 Activada por Mitógenos/antagonistas & inhibidores , Proteína Quinasa 1 Activada por Mitógenos/metabolismo , Proteína Quinasa 3 Activada por Mitógenos/antagonistas & inhibidores , Proteína Quinasa 3 Activada por Mitógenos/metabolismo , Miocardio/patología , Miofibroblastos/efectos de los fármacos , Miofibroblastos/metabolismo , Miofibroblastos/patología , Osteopontina/metabolismo , Inhibidores de Proteínas Quinasas/farmacología , Inhibidores de Proteínas Quinasas/uso terapéutico , Proteínas Serina-Treonina Quinasas/genéticaRESUMEN
A cladding-pumped 4-core erbium-doped fiber (4C-EDF) with a pedestal structure has been firstly, to the best of our knowledge, proposed and fabricated for space division multiplexing (SDM) amplification. The numerical simulation shows that the index-raised pedestal around the fiber core can improve power conversion efficiency (PCE) by enhancing pump power usage. Compared with conventional 4C-EDF, the 4C-EDF with a pedestal has a gain improvement of 4.5 dB and a PCE enhancement of 91.8%, according to the experimental results (pedestal fiber: 9.55%, conventional fiber: 4.98%). For a 6 dBm total input signal power at L-band and a 7.8 W pump power at 976 nm, the pedestal 4C-EDF shows an average gain of 25 dB and an average noise figure (NF) of 6.5 dB over all cores in the wavelength range of 1570.41 nm to 1610.87 nm. The core-to-core gain variation is less than 2 dB.
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OBJECTIVE: The superoxide-generating Nox2 (NADPH oxidase-2) is expressed in multiple cell types. Previous studies demonstrated distinct roles for cardiomyocyte, endothelial cell, and leukocyte cell Nox2 in ANG II (angiotensin II)-induced cardiovascular remodeling. However, the in vivo role of fibroblast Nox2 remains unclear. Approach and Results: We developed a novel mouse model with inducible fibroblast-specific deficiency of Nox2 (fibroblast-specific Nox2 knockout or Fibro-Nox2KO mice) and investigated the responses to chronic ANG II stimulation. Fibro-Nox2KO mice showed no differences in basal blood pressure or vessel wall morphology, but the hypertensive response to ANG II infusion (1.1 mg/[kg·day] for 14 days) was substantially reduced as compared to control Nox2-Flox littermates. This was accompanied by a significant attenuation of aortic and resistance vessel remodeling. The conditioned medium of ANG II-stimulated primary fibroblasts induced a significant increase in vascular smooth muscle cell growth, which was inhibited by the short hairpin RNA (shRNA)-mediated knockdown of fibroblast Nox2. Mass spectrometric analysis of the secretome of ANG II-treated primary fibroblasts identified GDF6 (growth differentiation factor 6) as a potential growth factor that may be involved in these effects. Recombinant GDF6 induced a concentration-dependent increase in vascular smooth muscle cell growth while chronic ANG II infusion in vivo significantly increased aortic GDF6 protein levels in control mice but not Fibro-Nox2KO animals. Finally, silencing GDF6 in fibroblasts prevented the induction of vascular smooth muscle cell growth by fibroblast-conditioned media in vitro. CONCLUSIONS: These results indicate that fibroblast Nox2 plays a crucial role in the development of ANG II-induced vascular remodeling and hypertension in vivo. Mechanistically, fibroblast Nox2 may regulate paracrine signaling to medial vascular smooth muscle cells via factors, such as GDF6.
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Fibroblastos/enzimología , Hipertensión/enzimología , Músculo Liso Vascular/metabolismo , Miocitos del Músculo Liso/metabolismo , NADPH Oxidasa 2/metabolismo , Comunicación Paracrina , Remodelación Vascular , Angiotensina II , Animales , Aorta/metabolismo , Aorta/patología , Aorta/fisiopatología , Presión Sanguínea , Células Cultivadas , Modelos Animales de Enfermedad , Factor 6 de Diferenciación de Crecimiento/genética , Factor 6 de Diferenciación de Crecimiento/metabolismo , Hipertensión/inducido químicamente , Hipertensión/genética , Hipertensión/patología , Masculino , Ratones Endogámicos C57BL , Ratones Noqueados , Músculo Liso Vascular/patología , Músculo Liso Vascular/fisiopatología , Miocitos del Músculo Liso/patología , NADPH Oxidasa 2/genética , Transducción de SeñalRESUMEN
In the heart, the serine carboxypeptidase cathepsin A (CatA) is distributed between lysosomes and the extracellular matrix (ECM). CatA-mediated degradation of extracellular peptides may contribute to ECM remodeling and left ventricular (LV) dysfunction. Here, we aimed to evaluate the effects of CatA overexpression on LV remodeling. A proteomic analysis of the secretome of adult mouse cardiac fibroblasts upon digestion by CatA identified the extracellular antioxidant enzyme superoxide dismutase (EC-SOD) as a novel substrate of CatA, which decreased EC-SOD abundance 5-fold. In vitro, both cardiomyocytes and cardiac fibroblasts expressed and secreted CatA protein, and only cardiac fibroblasts expressed and secreted EC-SOD protein. Cardiomyocyte-specific CatA overexpression and increased CatA activity in the LV of transgenic mice (CatA-TG) reduced EC-SOD protein levels by 43%. Loss of EC-SOD-mediated antioxidative activity resulted in significant accumulation of superoxide radicals (WT, 4.54 µmol/mg tissue/min; CatA-TG, 8.62 µmol/mg tissue/min), increased inflammation, myocyte hypertrophy (WT, 19.8 µm; CatA-TG, 21.9 µm), cellular apoptosis, and elevated mRNA expression of hypertrophy-related and profibrotic marker genes, without affecting intracellular detoxifying proteins. In CatA-TG mice, LV interstitial fibrosis formation was enhanced by 19%, and the type I/type III collagen ratio was shifted toward higher abundance of collagen I fibers. Cardiac remodeling in CatA-TG was accompanied by an increased LV weight/body weight ratio and LV end diastolic volume (WT, 50.8 µl; CatA-TG, 61.9 µl). In conclusion, CatA-mediated EC-SOD reduction in the heart contributes to increased oxidative stress, myocyte hypertrophy, ECM remodeling, and inflammation, implicating CatA as a potential therapeutic target to prevent ventricular remodeling.
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Catepsina A/metabolismo , Miocitos Cardíacos/metabolismo , Proteolisis , Superóxido Dismutasa/metabolismo , Remodelación Ventricular , Animales , Catepsina A/genética , Masculino , Ratones , Ratones Transgénicos , Miocitos Cardíacos/patología , Superóxido Dismutasa/genéticaRESUMEN
The gain bandwidth of the erbium-doped fiber amplifier limits the enhancement of the transmission capacity in optical fiber communication systems. This Letter reports an erbium-ytterbium co-doped phosphosilicate fiber, which is expected to increase transmission capacity by extending the L-band gain bandwidth to 1623 nm. The fiber was fabricated by modified chemical vapor deposition combined with solution doping technology. The mechanism of bandwidth-expansion by inhibiting the signal excited-state absorption was investigated. When the signal power and pump power were maintained at -3.7dBm and â¼720mW at 1480 nm, the 20 dB gain range was extended out to 1623 nm. Additionally, the noise figure at 1623 nm decreased to 6.01 dB, with 23 dBm saturated output power. The results show that the erbium-ytterbium co-doped phosphosilicate fiber has a great potential for extending L-band amplification.
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BACKGROUND: Premature birth entails an adverse cardiovascular risk profile, but the underlying mechanisms are insufficiently understood. Here, we employed an unbiased cardiovascular proteomics approach to profile former very preterm-born preschoolers. METHODS: This observational study investigated differences in plasma concentrations of 79 proteins, including putative cardiovascular biomarkers between very preterm- and term-born children on average 5.5 years old (53.1% male) using multiple-reaction monitoring mass spectrometry. RESULTS: Very preterm-born (n = 38; median gestational age 29.6 weeks) compared to term-born (n = 26; 40.2 weeks) children featured lower plasma concentrations of platelet factor 4 (PLF4; -61.6%, P < 0.0001), platelet basic protein (CXCL7; -57.8%, P < 0.0001), and hemoglobin subunit beta (-48.3%, P < 0.0001). Results remained virtually unchanged when adjusting for complete blood count parameters, including platelet count. Conversely, whole blood hemoglobin was higher (+7.62%, P < 0.0001) in preterm-born children. CONCLUSIONS: Very preterm birth was associated with decreased markers of platelet activation among preschoolers. These findings are consistent with reduced platelet reactivity persisting from very preterm birth to a preschool age. IMPACT: Former very preterm-born preschoolers featured reduced levels of platelet activation markers. While lower platelet reactivity in very preterm-born compared to term-born infants in the first days of life was established, it was unknown when, if at all, reactivity normalizes. The current study suggests that platelet hyporeactivity due to very preterm birth persists at least up to a preschool age. "Immaturity of the hemostatic system" may be a persistent sequel of preterm birth, but larger studies are needed to investigate its potential clinical implications.
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Enfermedades del Prematuro/sangre , Activación Plaquetaria , Nacimiento Prematuro/sangre , Biomarcadores , Sistema Cardiovascular , Preescolar , Femenino , Edad Gestacional , Humanos , Recién Nacido , Recien Nacido Prematuro , Masculino , Análisis Multivariante , Proteómica/métodos , Riesgo , Factores de RiesgoRESUMEN
BACKGROUND: Extracellular matrix (ECM) remodeling contributes to in-stent restenosis and thrombosis. Despite its important clinical implications, little is known about ECM changes post-stent implantation. METHODS: Bare-metal and drug-eluting stents were implanted in pig coronary arteries with an overstretch under optical coherence tomography guidance. Stented segments were harvested 1, 3, 7, 14, and 28 days post-stenting for proteomics analysis of the media and neointima. RESULTS: A total of 151 ECM and ECM-associated proteins were identified by mass spectrometry. After stent implantation, proteins involved in regulating calcification were upregulated in the neointima of drug-eluting stents. The earliest changes in the media were proteins involved in inflammation and thrombosis, followed by changes in regulatory ECM proteins. By day 28, basement membrane proteins were reduced in drug-eluting stents in comparison with bare-metal stents. In contrast, the large aggregating proteoglycan aggrecan was increased. Aggrecanases of the ADAMTS (a disintegrin and metalloproteinase with thrombospondin motifs) family contribute to the catabolism of vascular proteoglycans. An increase in ADAMTS-specific aggrecan fragments was accompanied by a notable shift from ADAMTS1 and ADAMTS5 to ADAMTS4 gene expression after stent implantation. Immunostaining in human stented coronary arteries confirmed the presence of aggrecan and aggrecan fragments, in particular, at the contacts of the stent struts with the artery. Further investigation of aggrecan presence in the human vasculature revealed that aggrecan and aggrecan cleavage were more abundant in human arteries than in human veins. In addition, aggrecan synthesis was induced on grafting a vein into the arterial circulation, suggesting an important role for aggrecan in vascular plasticity. Finally, lack of ADAMTS-5 activity in mice resulted in an accumulation of aggrecan and a dilation of the thoracic aorta, confirming that aggrecanase activity regulates aggrecan abundance in the arterial wall and contributes to vascular remodeling. CONCLUSIONS: Significant differences were identified by proteomics in the ECM of coronary arteries after bare-metal and drug-eluting stent implantation, most notably an upregulation of aggrecan, a major ECM component of cartilaginous tissues that confers resistance to compression. The accumulation of aggrecan coincided with a shift in ADAMTS gene expression. This study provides the first evidence implicating aggrecan and aggrecanases in the vascular injury response after stenting.
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Proteínas ADAMTS/metabolismo , Agrecanos , Vasos Coronarios/cirugía , Endopeptidasas/metabolismo , Matriz Extracelular/enzimología , Intervención Coronaria Percutánea/instrumentación , Proteómica/métodos , Stents , Remodelación Vascular , Proteínas ADAMTS/genética , Proteína ADAMTS5/genética , Proteína ADAMTS5/metabolismo , Animales , Cromatografía Líquida de Alta Presión , Vasos Coronarios/enzimología , Vasos Coronarios/fisiopatología , Stents Liberadores de Fármacos , Endopeptidasas/genética , Femenino , Humanos , Masculino , Metales , Ratones Noqueados , Modelos Animales , Neointima , Diseño de Prótesis , Transducción de Señal , Sus scrofa , Espectrometría de Masas en Tándem , Factores de TiempoRESUMEN
OBJECTIVE: Thoracic aortic aneurysm (TAA), a degenerative disease of the aortic wall, is accompanied by changes in the structure and composition of the aortic ECM (extracellular matrix). The ADAMTS (a disintegrin and metalloproteinase with thrombospondin motifs) family of proteases has recently been implicated in TAA formation. This study aimed to investigate the contribution of ADAMTS-5 to TAA development. APPROACH AND RESULTS: A model of aortic dilatation by AngII (angiotensin II) infusion was adopted in mice lacking the catalytic domain of ADAMTS-5 (Adamts5Δcat). Adamts5Δcat mice showed an attenuated rise in blood pressure while displaying increased dilatation of the ascending aorta (AsAo). Interestingly, a proteomic comparison of the aortic ECM from AngII-treated wild-type and Adamts5Δcat mice revealed versican as the most upregulated ECM protein in Adamts5Δcat mice. This was accompanied by a marked reduction of ADAMTS-specific versican cleavage products (versikine) and a decrease of LRP1 (low-density lipoprotein-related protein 1). Silencing LRP1 expression in human aortic smooth muscle cells reduced the expression of ADAMTS5, attenuated the generation of versikine, but increased soluble ADAMTS-1. A similar increase in ADAMTS-1 was observed in aortas of AngII-treated Adamts5Δcat mice but was not sufficient to maintain versican processing and prevent aortic dilatation. CONCLUSIONS: Our results support the emerging role of ADAMTS proteases in TAA. ADAMTS-5 rather than ADAMTS-1 is the key protease for versican regulation in murine aortas. Further studies are needed to define the ECM substrates of the different ADAMTS proteases and their contribution to TAA formation.
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Proteína ADAMTS5/metabolismo , Aorta Torácica/enzimología , Aneurisma de la Aorta Torácica/enzimología , Matriz Extracelular/enzimología , Remodelación Vascular , Proteína ADAMTS1/metabolismo , Proteína ADAMTS5/deficiencia , Proteína ADAMTS5/genética , Angiotensina II , Animales , Aorta Torácica/patología , Aneurisma de la Aorta Torácica/inducido químicamente , Aneurisma de la Aorta Torácica/genética , Aneurisma de la Aorta Torácica/patología , Células Cultivadas , Dilatación Patológica , Modelos Animales de Enfermedad , Matriz Extracelular/patología , Humanos , Proteína 1 Relacionada con Receptor de Lipoproteína de Baja Densidad/genética , Proteína 1 Relacionada con Receptor de Lipoproteína de Baja Densidad/metabolismo , Masculino , Ratones Noqueados , Músculo Liso Vascular/enzimología , Miocitos del Músculo Liso , Receptores de LDL/metabolismo , Proteínas Supresoras de Tumor/metabolismo , Versicanos/metabolismoRESUMEN
Aims: Human and mouse cardiac beta3-adrenergic receptors (beta3AR) exert antipathetic effects to those of beta1-2AR stimulation. We examined their role in modulating myocardial remodelling, particularly fibrosis in response to haemodynamic stress. Methods and results: Mice with cardiac myocyte-specific expression of beta3AR (ADRB3-tg) or tamoxifen-inducible homozygous deletion (c-Adrb3-ko, with loxP-targeted Adrb3) were submitted to transaortic constriction. A superfusion assay was used for proteomic analysis of paracrine mediators between beta3AR-expressing cardiac myocytes and cardiac fibroblasts cultured separately. We show that cardiac beta3AR attenuate myocardial fibrosis in response to haemodynamic stress. Interstitial fibrosis and collagen content were reduced in ADRB3-tg, but augmented in c-Adrb3-ko. ADRB3 and collagen (COL1A1) expression were also inversely related in ventricular biopsies of patients with valve disease. Incubation of cardiac fibroblasts with media conditioned by hypertrophic myocytes induced fibroblast proliferation, myo-differentiation, and collagen production. These effects were abrogated upon ADRB3 expression in myocytes. Comparative shotgun proteomic analysis of the myocyte secretomes revealed a number of factors differentially regulated by beta3AR, among which connective tissue growth factor [CTGF (CCN2)] was prominently reduced. CTGF was similarly reduced in stressed hearts from ADRB3-tg, but increased in hearts from c-Adrb3-ko mice. CTGF expression was mediated by reactive oxygen species production which was reduced by ADRB3 expression in vitro and in vivo. This antioxidant and anti-fibrotic effect involved beta3AR coupling to the neuronal isoform of nitric oxide synthase (nNOS) in cardiac myocytes, as both were abrogated upon nNOS inhibition or Nos1 homozygous deletion. Conclusion: Cardiac beta3AR protect from fibrosis in response to haemodynamic stress by modulating nitric oxide and oxidant stress-dependent paracrine signaling to fibroblasts. Specific agonism at beta3AR may offer a new therapeutic modality to prevent cardiac fibrosis.
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Fibrosis , Cardiopatías , Miocitos Cardíacos , Estrés Oxidativo/fisiología , Comunicación Paracrina/fisiología , Receptores Adrenérgicos beta 3/metabolismo , Animales , Catecolaminas/metabolismo , Fibrosis/metabolismo , Fibrosis/prevención & control , Cardiopatías/metabolismo , Cardiopatías/prevención & control , Ratones , Miocitos Cardíacos/citología , Miocitos Cardíacos/metabolismo , Óxido Nítrico/metabolismoRESUMEN
RATIONALE: Platelets shed microRNAs (miRNAs). Plasma miRNAs change on platelet inhibition. It is unclear whether plasma miRNA levels correlate with platelet function. OBJECTIVE: To link small RNAs to platelet reactivity. METHODS AND RESULTS: Next-generation sequencing of small RNAs in plasma revealed 2 peaks at 22 to 23 and 32 to 33 nucleotides corresponding to miRNAs and YRNAs, respectively. Among YRNAs, predominantly, fragments of RNY4 and RNY5 were detected. Plasma miRNAs and YRNAs were measured in 125 patients with a history of acute coronary syndrome who had undergone detailed assessment of platelet function 30 days after the acute event. Using quantitative real-time polymerase chain reactions, 92 miRNAs were assessed in patients with acute coronary syndrome on different antiplatelet therapies. Key platelet-related miRNAs and YRNAs were correlated with platelet function tests. MiR-223 (rp=0.28; n=121; P=0.002), miR-126 (rp=0.22; n=121; P=0.016), and other abundant platelet miRNAs and YRNAs showed significant positive correlations with the vasodilator-stimulated phosphoprotein phosphorylation assay. YRNAs, miR-126, and miR-223 were also among the small RNAs showing the greatest dependency on platelets and strongly correlated with plasma levels of P-selectin, platelet factor 4, and platelet basic protein in the population-based Bruneck study (n=669). A single-nucleotide polymorphism that facilitates processing of pri-miR-126 to mature miR-126 accounted for a rise in circulating platelet activation markers. Inhibition of miR-126 in mice reduced platelet aggregation. MiR-126 directly and indirectly affects ADAM9 and P2Y12 receptor expression. CONCLUSIONS: Levels of platelet-related plasma miRNAs and YRNAs correlate with platelet function tests in patients with acute coronary syndrome and platelet activation markers in the general population. Alterations in miR-126 affect platelet reactivity.
Asunto(s)
Síndrome Coronario Agudo/sangre , Plaquetas/metabolismo , MicroARNs/sangre , Activación Plaquetaria , Síndrome Coronario Agudo/tratamiento farmacológico , Síndrome Coronario Agudo/genética , Animales , Plaquetas/efectos de los fármacos , Línea Celular Tumoral , Perfilación de la Expresión Génica/métodos , Humanos , Masculino , Ratones , Ratones Endogámicos C57BL , MicroARNs/genética , Oligonucleótidos/genética , Oligonucleótidos/metabolismo , Activación Plaquetaria/efectos de los fármacos , Activación Plaquetaria/genética , Inhibidores de Agregación Plaquetaria/uso terapéutico , Pruebas de Función Plaquetaria , Polimorfismo de Nucleótido Simple , Reacción en Cadena en Tiempo Real de la Polimerasa , TransfecciónRESUMEN
Aims: Post-operative atrial fibrillation (POAF) is a predictor of morbidity and mortality after cardiac surgery. Latent predisposing factors may reside in the epicardial adipose tissue (EAT) due to its anatomical position and high protein production rate. In order to explore a possible mechanistic link, we characterized proteins secreted by the EAT preceding the onset of POAF. Methods and results: Epicardial adipose tissue samples were collected from 76 consecutive patients with no history of AF undergoing coronary artery bypass surgery, 50 samples for proteomic analysis and 26 for gene expression studies, further divided according to development of POAF. Ten vs. 10 matched samples representing EAT secretome were analysed by two-dimensional difference in-gel electrophoresis (2D-DIGE) and high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) to identify differentially expressed proteins (P < 0.05, expression change >1.2 fold). Findings were validated by Western blotting on EAT protein extracts and by gene expression studies via quantitative polymerase chain reaction (qPCR). Proteomics returned 35 differentially expressed proteins. Amongst those, gelsolin was down regulated in POAF. Western blot analysis confirmed a significant reduction in gelsolin in the AF group. Gene expression for gelsolin was significantly reduced in the AF group confirming the proteomics findings. Conclusion: For the first time we describe EAT secretome as a possible substrate for POAF. It contains various proteins differentially expressed in patients who later develop POAF. Amongst those gelsolin, involved in inflammation and ion channel regulation, was associated with maintenance of sinus rhythm. Understanding the role of EAT may offer novel insights into prevention and treatment of AF.