Editorial: Myocardial fibrosis: What we know now.

Worldwide, cardiovascular diseases affect millions, cause serious economic burdens, and represent the number one cause of death. A broad range of pathological cardiac conditions is associated with myocardial tissue remodeling and fibrosis. Cardiac fibrosis reflects the exaggerated accumulation of extracellular matrix components, and activation of stromal cell compartments in the tissue, followed by acute or chronic inflammatory responses (see the Figure 1). Progressive cardiac fibrosis has been recognized to cause life-threatening arrhythmias. The development of life-saving therapeutic strategies and new medications requires extensive scientific efforts to understand the pathophysiology of pro-arrhythmogenic fibrosis, which is currently poorly understood. Understanding the cellular and molecular roots of cardiac fibrogenesis is crucial for identifying potential diagnostic and therapeutic targets in cardiovascular diseases. The aim of this Research Topic is a wide-ranging overview of the current understanding of the mechanisms of myocardial fibrosis across diverse cardiovascular disorders and its evaluation in patients

Stem Cells in Heart Failure

link_to_subscribed_fulltex

Nanokapsuły typu rdzeń-otoczka na bazie chitozanu

N-dodecyl derivative of cationically modified chitosan was used to prepare core-shell nanocapsules templated on liquid cores. Surfactant-free method based on ultrasound-assisted direct emulsification of aqueous solution of polysaccharide with oleic acid was applied. Formation of spherical capsules was confirmed by scanning and transmission electron microscopies. Dynamic light scattering measurements were used to determine physicochemical parameters of the obtained particles as well as to follow the process of multilayer shell formation. Confocal microscopy was applied to examine the ability of encapsulation of hydrophobic compounds inside the cores of the nanocapsules. Performed studies confirmed that hydrophobically modified cationic chitosan provides long-term stabilization of oil-in-water emulsion for biomedical applications as no toxic effect was observed in acute oral toxicity studies.Do przygotowania nanokapsuł na ciekłych rdzeniach stabilizowanych bez użycia małocząsteczkowych surfaktantów użyto N-dodecylowej pochodnej zmodyfikowanego kationowo chitozanu. Kapsuły otrzymano w procesie wspomaganej ultradźwiękami bezpośredniej emulsyfikacji fazy wodnej zawierającej modyfikowany polisacharyd oraz kwas oleinowy. Powstawanie sferycznych kapsuł potwierdzono za pomocą skaningowej oraz transmisyjnej mikroskopii elektronowej. Obrazowanie z użyciem mikroskopii konfokalnej posłużyło natomiast do zbadania zdolności do enkapsulacji hydrofobowych barwników w rdzeniach chitozanowych nanokapsuł. Stosując technikę dynamicznego rozpraszania światła wyznaczono fizykochemiczne parametry nanoemulsji oraz stwierdzono powstawanie wielowarstwowych otoczek. Przeprowadzone badania dowiodły, że zastosowanie hydrofobowo zmodyfikowanej kationowej pochodnej chitozanu pozwala na uzyskanie stabilnych w czasie emulsji typu olej w wodzie. Wykazany brak toksyczności układów w warunkach in vivo pozwala na ich zastosowanie do celów biomedycznych

Stable aqueous dispersion of superparamagnetic iron oxide nanoparticles protected by charged chitosan derivatives

This article presents the synthesis and characterization of biocompatible superparamagnetic iron oxide nanoparticles (SPIONs) coated with ultrathin layer of anionic derivative of chitosan. The water-based fabrication involved a two-step procedure. In the first step, the nanoparticles were obtained by co-precipita- tion of ferrous and ferric aqueous salt solutions with ammonia in the presence of cationic derivative of chitosan. In the second step, such prepared materials were subjected to adsorption of oppositely charged chitosan derivative which resulted in the preparation of negatively charged SPIONs. They were found to develop highly stable dispersion in water. The core size of the nanocoated SPIONs, determined using transmis- sion electron microscopy, was measured to be slightly above 10 nm. The coated nanoparticles form aggregates with majority of them having hydrodynamic diameter below 100 nm, as measured by dynamic light scattering. Their composition and properties were studied using FTIR and thermogravimetric analyses. They exhibit magnetic properties typical for superparamagnetic material with a high saturation magnetization value of 123 ± 12 emu g - 1 Fe. Very high value of the measured r 2 relaxivity, 369 ± 3mM - 1 s - 1 , is conducive for the potential application of the obtained SPIONs as prom- ising contrast agents in magnetic resonance imaging

T Lymphocyte-Derived Exosomes Transport MEK1/2 and ERK1/2 and Induce NOX4-Dependent Oxidative Stress in Cardiac Microvascular Endothelial Cells

Background: Activation of endothelial cells by inflammatory mediators secreted by CD4+ T lymphocytes plays a key role in the inflammatory response. Exosomes represent a specific class of signaling cues transporting a mixture of proteins, nucleic acids, and other biomolecules. So far, the impact of exosomes shed by T lymphocytes on cardiac endothelial cells remained unknown. Methods and results: Supernatants of CD4+ T cells activated with anti-CD3/CD28 beads were used to isolate exosomes by differential centrifugation. Activation of CD4+ T cells enhanced exosome production, and these exosomes (CD4-exosomes) induced oxidative stress in cardiac microvascular endothelial cells (cMVECs) without affecting their adhesive properties. Furthermore, CD4-exosome treatment aggravated the generation of mitochondrial reactive oxygen species (ROS), reduced nitric oxide (NO) levels, and enhanced the proliferation of cMVECs. These effects were reversed by adding the antioxidant apocynin. On the molecular level, CD4-exosomes increased NOX2, NOX4, ERK1/2, and MEK1/2 in cMVECs, and ERK1/2 and MEK1/2 proteins were found in CD4-exosomes. Inhibition of either MEK/ERK with U0126 or ERK with FR180204 successfully protected cMVECs from increased ROS levels and reduced NO bioavailability. Treatment with NOX1/4 inhibitor GKT136901 effectively blocked excessive ROS and superoxide production, reversed impaired NO levels, and reversed enhanced cMVEC proliferation triggered by CD4-exosomes. The siRNA-mediated silencing of Nox4 in cMVECs confirmed the key role of NOX4 in CD4-exosome-induced oxidative stress. To address the properties of exosomes under inflammatory conditions, we used the mouse model of CD4+ T cell-dependent experimental autoimmune myocarditis. In contrast to exosomes obtained from control hearts, exosomes obtained from inflamed hearts upregulated NOX2, NOX4, ERK1/2, MEK1/2, increased ROS and superoxide levels, and reduced NO bioavailability in treated cMVECs, and these changes were reversed by apocynin. Conclusion: Our results point to exosomes as a novel class of bioactive factors secreted by CD4+ T cells in immune response and represent potential important triggers of NOX4-dependent endothelial dysfunction. Neutralization of the prooxidative aspect of CD4-exosomes could open perspectives for the development of new therapeutic strategies in inflammatory cardiovascular diseases

The AP-1 transcription factor Fosl-2 regulates autophagy in cardiac fibroblasts during myocardial fibrogenesis

Background: Pathological activation of cardiac fibroblasts is a key step in development and progression of cardiac fibrosis and heart failure. This process has been associated with enhanced autophagocytosis, but molecular mechanisms remain largely unknown. Methods and Results: Immunohistochemical analysis of endomyocardial biopsies showed increased activation of autophagy in fibrotic hearts of patients with inflammatory cardiomyopathy. In vitro experiments using mouse and human cardiac fibroblasts confirmed that blockade of autophagy with Bafilomycin A1 inhibited fibroblast-to-myofibroblast transition induced by transforming growth factor (TGF)-β. Next, we observed that cardiac fibroblasts obtained from mice overexpressing transcription factor Fos-related antigen 2 (Fosl-2tg) expressed elevated protein levels of autophagy markers: the lipid modified form of microtubule-associated protein 1A/1B-light chain 3B (LC3BII), Beclin-1 and autophagy related 5 (Atg5). In complementary experiments, silencing of Fosl-2 with antisense GapmeR oligonucleotides suppressed production of type I collagen, myofibroblast marker alpha smooth muscle actin and autophagy marker Beclin-1 in cardiac fibroblasts. On the other hand, silencing of either LC3B or Beclin-1 reduced Fosl-2 levels in TGF-β-activated, but not in unstimulated cells. Using a cardiac hypertrophy model induced by continuous infusion of angiotensin II with osmotic minipumps, we confirmed that mice lacking either Fosl-2 (Ccl19CreFosl2flox/flox) or Atg5 (Ccl19CreAtg5flox/flox) in stromal cells were protected from cardiac fibrosis. Conclusion: Our findings demonstrate that Fosl-2 regulates autophagocytosis and the TGF-β-Fosl-2-autophagy axis controls differentiation of cardiac fibroblasts. These data provide a new insight for the development of pharmaceutical targets in cardiac fibrosis

TNF-α protects from exacerbated myocarditis and cardiac death by suppressing expansion of activated heart-reactive CD4+ T cells

BACKGROUND AND AIMS Tumour necrosis factor α (TNF-α) represents a classical proinflammatory cytokine and its increased levels positively correlate with the severity of many cardiovascular diseases. Surprisingly, some heart failure patients receiving high doses of anti- TNF-α antibodies showed serious health worsening. This work aimed to examine the role of TNF-α signalling on the development and progression of myocarditis and heart-specific autoimmunity. METHODS AND RESULTS Mice with genetic deletion of TNF-α (Tnf+/- and Tnf-/-) and littermate controls (Tnf+/+) were used to study myocarditis in the inducible and the transgenic T cell receptor (TCR-M) models. Tnf+/- and Tnf-/- mice immunized with α-myosin heavy chain peptide (αMyHC) showed reduced myocarditis incidence but the susceptible animals developed extensive inflammation in the heart. In the TCR-M model, defective TNF-α production was associated with increased mortality at a young age due to cardiomyopathy and cardiac fibrosis. We could confirm that TNF-α as well as the secretome of antigen-activated heart-reactive effector CD4+ T (Teff) cells effectively activated the adhesive properties of cardiac microvascular endothelial cells (cMVECs). Our data suggested that TNF-α produced by endothelial in addition to Teff cells promoted leucocyte adhesion to activated cMVECs. Analysis of CD4+ T lymphocytes from both models of myocarditis showed a strongly increased fraction of Teff cells in hearts, spleens, and in the blood of Tnf+/- and Tnf-/- mice. Indeed, antigen-activated Tnf-/- Teff cells showed prolonged long-term survival and TNF-α cytokine-induced cell death of heart-reactive Teff. CONCLUSIONS TNF-α signalling promotes myocarditis development by activating cardiac endothelial cells. However, in the case of established disease, TNF-α protects from exacerbating cardiac inflammation by inducing activation-induced cell death of heart-reactive Teff. These data might explain the lack of success of standard anti-TNF-α therapy in heart failure patients and open perspectives for T cell-targeted approaches

Nitric oxide synthase 2 is required for conversion of pro-fibrogenic inflammatory CD133+ progenitors into F4/80+ macrophages in experimental autoimmune myocarditis

Aims Experimental autoimmune myocarditis (EAM) model mirrors important mechanisms of inflammatory dilated cardiomyopathy (iDCM). In EAM, inflammatory CD133+ progenitors are a major cellular source of cardiac myofibroblasts in the post-inflammatory myocardium. We hypothesized that exogenous delivery of macrophage-colony-stimulating factor (M-CSF) can stimulate macrophage lineage differentiation of inflammatory progenitors and, therefore, prevent their naturally occurring myofibroblast fate in EAM. Methods and results EAM was induced in wild-type (BALB/c) and nitric oxide synthase 2-deficient (Nos2−/−) mice and CD133+ progenitors were isolated from inflamed hearts. In vitro, M-CSF converted inflammatory CD133+ progenitors into nitric oxide-producing F4/80+ macrophages and prevented transforming growth factor-β-mediated myofibroblast differentiation. Importantly, only a subset of heart-infiltrating CD133+ progenitors expresses macrophage-specific antigen F4/80 in EAM. These CD133+/F4/80hi cells show impaired myofibrogenic potential compared with CD133+/F4/80− cells. M-CSF treatment of wild-type mice with EAM at the peak of disease markedly increased CD133+/F4/80hi cells in the myocardium, and CD133+ progenitors isolated from M-CSF-treated mice failed to differentiate into myofibroblasts. In contrast, M-CSF was not effective in converting CD133+ progenitors from inflamed hearts of Nos2−/− mice into macrophages, and M-CSF treatment did not result in increased CD133+/F4/80hi cell population in hearts of Nos2−/− mice. Accordingly, M-CSF prevented post-inflammatory fibrosis and left ventricular dysfunction in wild-type but not in Nos2−/− mice. Conclusion Active and NOS2-dependent induction of macrophage lineage differentiation abrogates the myofibrogenic potential of heart-infiltrating CD133+ progenitors. Modulating the in vivo differentiation fate of specific progenitors might become a novel approach for the treatment of inflammatory heart disease