RATIONALE: Cardiac ECM (extracellular matrix) comprises a dynamic molecular network providing structural support to heart tissue function. Understanding the impact of ECM remodeling on cardiac cells during heart failure (HF) is essential to prevent adverse ventricular remodeling and restore organ functionality in affected patients. OBJECTIVES: We aimed to (1) identify consistent modifications to cardiac ECM structure and mechanics that contribute to HF and (2) determine the underlying molecular mechanisms. METHODS AND RESULTS: We first performed decellularization of human and murine ECM (decellularized ECM) and then analyzed the pathological changes occurring in decellularized ECM during HF by atomic force microscopy, 2-photon microscopy, high-resolution 3-dimensional image analysis, and computational fluid dynamics simulation. We then performed molecular and functional assays in patient-derived cardiac fibroblasts based on YAP (yes-associated protein)-transcriptional enhanced associate domain (TEAD) mechanosensing activity and collagen contraction assays. The analysis of HF decellularized ECM resulting from ischemic or dilated cardiomyopathy, as well as from mouse infarcted tissue, identified a common pattern of modifications in their 3-dimensional topography. As compared with healthy heart, HF ECM exhibited aligned, flat, and compact fiber bundles, with reduced elasticity and organizational complexity. At the molecular level, RNA sequencing of HF cardiac fibroblasts highlighted the overrepresentation of dysregulated genes involved in ECM organization, or being connected to TGFβ1 (transforming growth factor β1), interleukin-1, TNF-α, and BDNF signaling pathways. Functional tests performed on HF cardiac fibroblasts pointed at mechanosensor YAP as a key player in ECM remodeling in the diseased heart via transcriptional activation of focal adhesion assembly. Finally, in vitro experiments clarified pathological cardiac ECM prevents cell homing, thus providing further hints to identify a possible window of action for cell therapy in cardiac diseases. CONCLUSIONS: Our multiparametric approach has highlighted repercussions of ECM remodeling on cell homing, cardiac fibroblast activation, and focal adhesion protein expression via hyperactivated YAP signaling during HF.
- MeSH
- adaptorové proteiny signální transdukční genetika metabolismus MeSH
- buněčný převod mechanických signálů MeSH
- dilatační kardiomyopatie genetika metabolismus patologie patofyziologie MeSH
- extracelulární matrix genetika metabolismus ultrastruktura MeSH
- fibroblasty metabolismus ultrastruktura MeSH
- funkce levé komory srdeční * MeSH
- infarkt myokardu genetika metabolismus patologie patofyziologie MeSH
- kultivované buňky MeSH
- lidé MeSH
- modely nemocí na zvířatech MeSH
- myokard metabolismus ultrastruktura MeSH
- myši inbrední C57BL MeSH
- pohyb buněk MeSH
- remodelace komor * MeSH
- srdeční selhání genetika metabolismus patologie patofyziologie MeSH
- studie případů a kontrol MeSH
- transkripční faktory genetika metabolismus MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- audiovizuální média MeSH
- časopisecké články MeSH
- práce podpořená grantem MeSH
This study is focused on the analysis of extracellular DNA (eDNA) from a biofilm matrix formed by Staphylococcus aureus, Listeria monocytogenes, and Salmonella enterica. The presence of eDNA in the biofilm of all the studied strains was confirmed by confocal laser scanning microscopy using fluorescent dyes with high affinity to nucleic acid. The protocol for eDNA isolation from the biofilm matrix was established, and subsequent characterization of the eDNA was performed. The purified eDNA obtained from the biofilm matrix of all three microorganisms was compared to the genomic DNA (gDNA) isolated from relevant planktonic grown cells. The process of eDNA isolation consisted of biofilm cultivation, its collection, sonication, membrane filtration, dialysis, lyophilisation, and extraction of DNA separated from the biofilm matrix with cetyltrimethylammonium bromide. An amplified fragment length polymorphism (AFLP) was used for comparing eDNA and gDNA. AFLP profiles showed a significant similarity between eDNA and gDNA at the strain level. The highest similarity, with a profile concordance rate of 94.7% per strain, was observed for S. aureus, L. monocytogenes, and S. enterica exhibited lower profiles similarity (78% and 60%, respectively). The obtained results support the hypothesis that the eDNA of studied bacterial species has its origin in the gDNA.
The lysyl oxidases (LOXs) are a family of enzymes deputed to cross-link collagen and elastin, shaping the structure and strength of the extracellular matrix (ECM). However, many novel "non-canonical" functions, alternative substrates, and regulatory mechanisms have been described and are being continuously elucidated. The activity of LOXs, therefore, appears to be integrated into a complex network of signals regulating many cell functions, including survival/proliferation/differentiation. Among these signaling pathways, TGF-β and PI3K/Akt/mTOR, in particular, cross-talk extensively with each other and with LOXs also initiating complex feedback loops which modulate the activity of LOXs and direct the remodeling of the ECM. A growing body of evidence indicates that LOXs are not only important in the homeostasis of the normal structure of the ECM, but are also implicated in the establishment and maturation of the tumor microenvironment. LOXs' association with advanced and metastatic cancer is well established; however, there is enough evidence to support a significant role of LOXs in the transformation of normal epithelial cells, in the accelerated tumor development and the induction of invasion of the premalignant epithelium. A better understanding of LOXs and their interactions with the different elements of the tumor immune microenvironment will prove invaluable in the design of novel anti-tumor strategies.
- MeSH
- biologické markery MeSH
- extracelulární matrix genetika metabolismus MeSH
- imunita * MeSH
- lidé MeSH
- lysyloxidasa chemie imunologie MeSH
- multigenová rodina MeSH
- nádorové mikroprostředí imunologie MeSH
- nádory etiologie patologie MeSH
- regulace genové exprese u nádorů MeSH
- signální transdukce MeSH
- vztahy mezi strukturou a aktivitou MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
This study presents a toxicological evaluation of two types of carbon dots (CD), similar in size (<10 nm) but differing in surface charge. Whole-genome mRNA and miRNA expression (RNAseq), as well as gene-specific DNA methylation changes, were analyzed in human embryonic lung fibroblasts (HEL 12469) after 4 h and 24 h exposure to concentrations of 10 and 50 µg/mL (for positive charged CD; pCD) or 10 and 100 µg/mL (for negative charged CD, nCD). The results showed a distinct response for the tested nanomaterials (NMs). The exposure to pCD induced the expression of a substantially lower number of mRNAs than those to nCD, with few commonly differentially expressed genes between the two CDs. For both CDs, the number of deregulated mRNAs increased with the dose and exposure time. The pathway analysis revealed a deregulation of processes associated with immune response, tumorigenesis and cell cycle regulation, after exposure to pCD. For nCD treatment, pathways relating to cell proliferation, apoptosis, oxidative stress, gene expression, and cycle regulation were detected. The expression of miRNAs followed a similar pattern: more pronounced changes after nCD exposure and few commonly differentially expressed miRNAs between the two CDs. For both CDs the pathway analysis based on miRNA-mRNA interactions, showed a deregulation of cancer-related pathways, immune processes and processes involved in extracellular matrix interactions. DNA methylation was not affected by exposure to any of the two CDs. In summary, although the tested CDs induced distinct responses on the level of mRNA and miRNA expression, pathway analyses revealed a potential common biological impact of both NMs independent of their surface charge.
- MeSH
- apoptóza účinky léků genetika MeSH
- exprese genu účinky léků genetika MeSH
- extracelulární matrix genetika MeSH
- fibroblasty účinky léků MeSH
- kultivované buňky MeSH
- lidé MeSH
- messenger RNA genetika MeSH
- metylace DNA účinky léků genetika MeSH
- mikro RNA genetika MeSH
- nádory genetika MeSH
- oxidační stres účinky léků genetika MeSH
- plíce účinky léků MeSH
- proliferace buněk účinky léků genetika MeSH
- signální transdukce účinky léků genetika MeSH
- stanovení celkové genové exprese metody MeSH
- uhlík farmakologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
In the adult brain, the extracellular matrix (ECM) influences recovery after injury, susceptibility to mental disorders, and is in general a strong regulator of neuronal plasticity. The proteoglycan aggrecan is a core component of the condensed ECM structures termed perineuronal nets (PNNs), and the specific role of PNNs on neural plasticity remains elusive. Here, we genetically targeted the Acan gene encoding for aggrecan using a novel animal model. This allowed for conditional and targeted loss of aggrecan in vivo, which ablated the PNN structure and caused a shift in the population of parvalbumin-expressing inhibitory interneurons toward a high plasticity state. Selective deletion of the Acan gene in the visual cortex of male adult mice reinstated juvenile ocular dominance plasticity, which was mechanistically identical to critical period plasticity. Brain-wide targeting improved object recognition memory.SIGNIFICANCE STATEMENT The study provides the first direct evidence of aggrecan as the main functional constituent and orchestrator of perineuronal nets (PNNs), and that loss of PNNs by aggrecan removal induces a permanent state of critical period-like plasticity. Loss of aggrecan ablates the PNN structure, resulting in invoked juvenile plasticity in the visual cortex and enhanced object recognition memory.
- MeSH
- agrekany analýza nedostatek genetika MeSH
- buněčné linie MeSH
- extracelulární matrix chemie genetika metabolismus MeSH
- myši inbrední C57BL MeSH
- myši knockoutované MeSH
- myši transgenní MeSH
- myši MeSH
- nervová síť chemie metabolismus MeSH
- neuroplasticita fyziologie MeSH
- světelná stimulace metody MeSH
- zrakové korové centrum chemie metabolismus MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
BACKGROUND: MicroRNAs are short non-coding regulators of gene expression. The human miR-29 family consists of three members: miR-29a, miR-29b and miR-29c. Members of this family were found to be aberrantly expressed in various types of tumors, including hematological malignancies. This family was described to have both oncogenic and tumor suppressor features influencing various pathological processes, such as tumor growth and apoptosis. This review summarizes current knowledge about the miR-29 family in selected hematological malignancies. CONCLUSION: Recent research of miR-29 family in hematological malignancies has proven its oncogenic as well as tumor suppressive potential. Nevertheless, the level of current evidence is not sufficient, and data remain inconclusive.
- MeSH
- apoptóza genetika MeSH
- extracelulární matrix genetika MeSH
- hematologické nádory genetika MeSH
- lidé MeSH
- mikro RNA genetika fyziologie MeSH
- nádorová transformace buněk genetika MeSH
- proliferace buněk genetika MeSH
- stárnutí buněk genetika MeSH
- tumor supresorové geny fyziologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
Paxillin (PXN) is a focal adhesion protein that has been implicated in signal transduction from the extracellular matrix. Recently, it has been shown to shuttle between the cytoplasm and the nucleus. When inside the nucleus, paxillin promotes cell proliferation. Here, we introduce paxillin as a transcriptional regulator of IGF2 and H19 genes. It does not affect the allelic expression of the two genes; rather, it regulates long-range chromosomal interactions between the IGF2 or H19 promoter and a shared distal enhancer on an active allele. Specifically, paxillin stimulates the interaction between the enhancer and the IGF2 promoter, thus activating IGF2 gene transcription, whereas it restrains the interaction between the enhancer and the H19 promoter, downregulating the H19 gene. We found that paxillin interacts with cohesin and the mediator complex, which have been shown to mediate long-range chromosomal looping. We propose that these interactions occur at the IGF2 and H19 gene cluster and are involved in the formation of loops between the IGF2 and H19 promoters and the enhancer, and thus the expression of the corresponding genes. These observations contribute to a mechanistic explanation of the role of paxillin in proliferation and fetal development.
- MeSH
- buňky Hep G2 MeSH
- chromozomální proteiny, nehistonové genetika MeSH
- extracelulární matrix genetika MeSH
- fokální adheze genetika MeSH
- genomový imprinting genetika MeSH
- insulinu podobný růstový faktor II biosyntéza genetika MeSH
- lidé MeSH
- metylace DNA genetika MeSH
- paxilin aplikace a dávkování MeSH
- proliferace buněk účinky léků genetika MeSH
- promotorové oblasti (genetika) MeSH
- proteiny buněčného cyklu genetika MeSH
- RNA dlouhá nekódující biosyntéza genetika MeSH
- signální transdukce účinky léků MeSH
- vývoj plodu genetika MeSH
- vývojová regulace genové exprese MeSH
- zesilovače transkripce MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
One of the generally recognized factors contributing to the initiation and maintenance of atrial fibrillation (AF) is structural remodeling of the myocardium that affects both atrial cardiomyocytes as well as interstitium. The goal of this study was to characterize morphologically and functionally interstitium of atria in patients with AF or in sinus rhythm (SR) who were indicated to heart surgery. Patient population consisted of 46 subjects (19 with long-term persistent AF, and 27 in SR) undergoing coronary bypass or valve surgery. Peroperative bioptic samples of the left and the right atria were examined using immunohistochemistry to visualize and quantify collagen I, collagen III, elastin, desmin, smooth muscle actin, endothelium and Vascular Endothelial Growth Factor (VEGF). The content of interstitial elastin, collagen I, and collagen III in atrial tissue was similar in AF and SR groups. However, the right atrium was more than twofold more abundant in elastin as compared with the left atrium and similar difference was found for collagen I and III. The right atrium showed also higher VEGF expression and lower microvascular density as compared to the left atrium. No significant changes in atrial extracellular matrix fiber content, microvascular density and angiogenic signaling, attributable to AF, were found in this cohort of patients with structural heart disease. This finding suggests that interstitial fibrosis and other morphological changes in atrial tissue are rather linked to structural heart disease than to AF per se. Significant regional differences in interstitial structure between right and left atrium is a novel observation that deserves further investigation.
- MeSH
- elastin genetika metabolismus MeSH
- extracelulární matrix genetika metabolismus MeSH
- fibrilace síní metabolismus patologie patofyziologie MeSH
- kolagen typ III genetika metabolismus MeSH
- kolagen typu I genetika metabolismus MeSH
- lidé středního věku MeSH
- lidé MeSH
- orgánová specificita MeSH
- senioři MeSH
- srdce - funkce síní MeSH
- srdeční frekvence * MeSH
- srdeční síně metabolismus patologie MeSH
- studie případů a kontrol MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The genetic cause of GAPO syndrome, a condition characterized by growth retardation, alopecia, pseudoanodontia, and progressive visual impairment, has not previously been identified. We studied four ethnically unrelated affected individuals and identified homozygous nonsense mutations (c.262C>T [p.Arg88*] and c.505C>T [p.Arg169*]) or splicing mutations (c.1435-12A>G [p.Gly479Phefs*119]) in ANTXR1, which encodes anthrax toxin receptor 1. The nonsense mutations predictably trigger nonsense-mediated mRNA decay, resulting in the loss of ANTXR1. The transcript with the splicing mutation theoretically encodes a truncated ANTXR1 containing a neopeptide composed of 118 unique amino acids in its C terminus. GAPO syndrome's major phenotypic features, which include dental abnormalities and the accumulation of extracellular matrix, recapitulate those found in Antxr1-mutant mice and point toward an underlying defect in extracellular-matrix regulation. Thus, we propose that mutations affecting ANTXR1 function are responsible for this disease's characteristic generalized defect in extracellular-matrix homeostasis.
- MeSH
- alopecie genetika patologie MeSH
- alternativní sestřih genetika MeSH
- anodoncie genetika patologie MeSH
- dědičné atrofie optického nervu genetika patologie MeSH
- DNA primery genetika MeSH
- extracelulární matrix genetika metabolismus MeSH
- fibroblasty MeSH
- fluorescenční protilátková technika MeSH
- frekvence genu MeSH
- genetická predispozice k nemoci genetika MeSH
- homeostáza genetika MeSH
- lidé MeSH
- lidské chromozomy, pár 2 genetika MeSH
- místa sestřihu RNA genetika MeSH
- molekulární sekvence - údaje MeSH
- nádorové proteiny genetika MeSH
- nesmyslný kodon genetika MeSH
- polymerázová řetězová reakce s reverzní transkripcí MeSH
- poruchy růstu genetika patologie MeSH
- receptory buněčného povrchu genetika MeSH
- rodokmen MeSH
- sekvence nukleotidů MeSH
- sekvenční analýza DNA MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- kazuistiky MeSH
- práce podpořená grantem MeSH
- Research Support, N.I.H., Intramural MeSH
- MeSH
- aneuploidie MeSH
- chromozomální aberace klasifikace MeSH
- diagnostické techniky molekulární metody MeSH
- DNA krev MeSH
- extracelulární matrix genetika MeSH
- fetální krev fyziologie metabolismus MeSH
- financování organizované MeSH
- komplikace těhotenství diagnóza krev MeSH
- lidé MeSH
- maternofetální výměna látek genetika MeSH
- novorozenecká aloimunitní trombocytopenie genetika MeSH
- prenatální diagnóza metody MeSH
- procesy určující pohlaví MeSH
- těhotenství MeSH
- Check Tag
- lidé MeSH
- těhotenství MeSH
- ženské pohlaví MeSH
