10 research outputs found
A Protective Role of FAM13A in Human Airway Epithelial Cells Upon Exposure to Cigarette Smoke Extract
Get PDFBACKGROUND: Chronic Obstructive Pulmonary Disease (COPD) is a progressive lung disease characterized by chronic inflammation upon inhalation of noxious particles, e.g., cigarette smoke. FAM13A is one of the genes often found to be associated with COPD, however its function in the pathophysiology of COPD is incompletely understood. We studied its role in airway epithelial barrier integrity and cigarette smoke-induced epithelial responses. MATERIALS AND METHODS: Protein level and localization of FAM13A was assessed with immunohistochemistry in lung tissue from COPD patients and non-COPD controls. In vitro, FAM13A expression was determined in the absence or presence of cigarette smoke extract (CSE) in primary airway epithelial cells (AECs) from COPD patients and controls by western blotting. FAM13A was overexpressed in cell line 16HBE14o- and its effect on barrier function was monitored real-time by electrical resistance. Expression of junctional protein E-cadherin and β-catenin was assessed by western blotting. The secretion of neutrophil attractant CXCL8 upon CSE exposure was measured by ELISA. RESULTS: FAM13A was strongly expressed in airway epithelium, but significantly weaker in airways of COPD patients compared to non-COPD controls. In COPD-derived AECs, but not those of controls, FAM13A was significantly downregulated by CSE. 16HBE14o- cells overexpressing FAM13A built up epithelial resistance significantly more rapidly, which was accompanied by higher E-cadherin expression and reduced CSE-induced CXCL8 levels. CONCLUSION: Our data indicate that the expression of FAM13A is lower in airway epithelium of COPD patients compared to non-COPD controls. In addition, cigarette smoking selectively downregulates airway epithelial expression of FAM13A in COPD patients. This may have important consequences for the pathophysiology of COPD, as the more rapid build-up of epithelial resistance upon FAM13A overexpression suggests improved (re)constitution of barrier function. The reduced epithelial secretion of CXCL8 upon CSE-induced damage suggests that lower FAM13A expression upon cigarette smoking may facilitate epithelial-driven neutrophilia
From Differential DNA Methylation in COPD to Mitochondria:Regulation of AHRR Expression Affects Airway Epithelial Response to Cigarette Smoke
Get PDFCigarette smoking causes hypomethylation of the gene Aryl Hydrocarbon Receptor Repressor (AHRR), which regulates detoxification and oxidative stress-responses. We investigated whether AHRR DNA methylation is related to chronic obstructive pulmonary disease (COPD) and studied its function in airway epithelial cells (AECs). The association with COPD was assessed in blood from never and current smokers with/without COPD, and in AECs from ex-smoking non-COPD controls and GOLD stage II-IV COPD patients cultured with/without cigarette smoke extract (CSE). The effect of CRISPR/Cas9-induced AHRR knockout on proliferation, CSE-induced mitochondrial membrane potential and apoptosis/necrosis in human bronchial epithelial 16HBE cells was studied. In blood, DNA methylation of AHRR at cg05575921 and cg21161138 was lower in smoking COPD subjects than smoking controls. In vitro, AHRR DNA methylation at these CpG-sites was lower in COPD-derived than control-derived AECs only upon CSE exposure. Upon AHRR knockout, we found a lower proliferation rate at baseline, stronger CSE-induced decrease in mitochondrial membrane potential, and higher CSE-induced late apoptosis/necroptosis. Together, our results show lower DNA methylation of AHRR upon smoking in COPD patients compared to non-COPD controls. Our data suggest that higher airway epithelial AHRR expression may lead to impaired cigarette smoke-induced mitochondrial dysfunction and apoptosis/necroptosis, potentially promoting unprogrammed/immunogenic cell death
Study of miRNA expression in invasive aspergillosis
No full textIntroduction: miRNAs are key regulators of various physiological and pathophysiological processes and their altered expression levels have been associated to many diseases. Increasing number of evidences have demonstrated that specific miRNAs can serve as novel biomarkers for clinical diagnosis of diseases. Invasive aspergillosis is life-threatening systemic fungal infection. Regrettably, the standard diagnostic methods for this condition take days to produce a reliable result.
Objectives: The aim of this study was to probe the expression profiles of 15 selected miRNAs in patients, with fevered neutropenia. We hypothesized that invasive aspergillosis could change the miRNA profiles in the blood. Our goal was to make a rapid test supporting diagnosis of invasive mycoses.
Methods: We obtained whole blood samples from 16 patients suffering neutropenic fever and from 5 healthy volunteers without neutropenic fever. Total RNA was isolated from the whole blood and were reverse transcribed to cDNA. TaqMan® quantitative real-time PCR (miRNA assay) was used for detecting miRNA expression profiles. Relative mRNA expressions were calculated with the ΔΔCt method.
Results: 7 out of the 19 miRNAs showed significant upregulation in the expression levels at p<0.05.
Conclusion: The miRNAs with significantly different expression between the test group and the control can be used as informative biomarkers to assess and monitor the body’s physio-pathological status in invasive aspergillosis.MSc/MAmolekuláris biológiaangolBiokémia-genomik
Accelerated ageing in COPD : epithelial barrier dysfunction as a driver of abnormal lung tissue repair and remodelling
No full textThe full abstract for this thesis is available in the body of the thesis, and will be available when the embargo expires.Medicine, Faculty ofAnesthesiology, Pharmacology and Therapeutics, Department ofGraduat
Palynomorph assemblage biozonation of Paleogene strata in Bende–Umuahia Area, Niger Delta Basin, southeastern Nigeria
No full textAbstractCenozoic sediments form extensive outcrops in the Niger Delta Basin. Detailed palynostratigraphic study was undertaken across Paleogene sequences exposed in Bende–Umuahia Area in up-dip sectors of the Niger Delta Basin, southeastern Nigeria, to establish different palynomorphs assemblage zones, with their corresponding ages. Palynological analysis was carried out on 27 selected outcrop samples, using the conventional maceration technique for recovering acid-insoluble organic-walled microfossils from sediments. Three Cenozoic lithostratigraphic units, including Imo Formation, Ameki Formation, and Ogwashi Formation, are exposed in the study area. Lithologies are sandstone, carbonaceous shale, mudstone, limestone, and the lignite. A total of 65 species of sporomorphs and 51 dinoflagellate cysts were identified. The recovered spores and pollen grains were used to establish six informal palynomorph assemblage zones, labeled as zone A – zone F, based on the first and the last occurrences of two or more species. These palynomorph assemblage zones include: (1) zone A — middle Paleocene Scabratriporites simpliformis–Bombacidites annae zone; (2) zone B — late Paleocene Foveotricolporites crassiexinus–Mauritidiites crassiexinus zone; (3) zone C — early Eocene Striatopollis catatumbus–Momipites africanus zone; (4) zone D — middle Eocene Margocolporites umuahiaensis–Gemmastephanocolporites brevicolpites zone; (5) zone E — late Eocene Cicatricosisporites dorogensis–Perfotricolpites nigerianus zone; and, (6) zone F — Oligocene–early Miocene Verrucatosporites usmensis–Magnastriatites howardii zone. The erected palynozones were correlated and compared with existing biozones in subsurface, down-dip sectors of the Niger Delta Basin, with pantropical palynological zones in tropical areas of Africa, and with palynofloral provinces of northern South America. A comparison of palynozones studied in southeastern Nigeria with other international palynozones, in this study, will assist in establishing the correlation of sediments for these areas.</jats:p
Free circulating mircoRNAs support the diagnosis of invasive aspergillosis in patients with hematologic malignancies and neutropenia
No full textAbstractFungal infections represent a worrisome complication in hematologic cancer patients and in the absence of disease specific symptoms, it is important to establish new biological indicators, which can be used during mould-active prophylaxis. Recently, miRNAs have appeared as candidate diagnostic and prognostic markers of several diseases. A pilot clinical study was performed to evaluate the diagnostic utility of 14 microRNAs which can be related to invasive fungal infections. Based on our data miR-142-3p, miR-142-5p, miR-26b-5p and miR-21-5p showed significant overexpression (p < 0.005) due to invasive aspergillosis in hemato-oncology patients with profound neutropenia. A tetramiR assay was designed to monitor peripheral blood specimens. Optimal cut-off was estimated by using the median value (fold change 1.1) of the log10 transformed gene expressions. The biomarker panel was evaluated on two independent sample cohorts implementing different antimicrobial prophylactic strategies. The receiver operating characteristic analysis with area under the curve proved to be 0.97. Three miRNAs (miR-142-5p, miR-142-3p, miR-16-5p) showed significant expression alterations in episodes with sepsis. In summary, the tetramiR assay proved to be a promising diagnostic adjunct with sufficient accuracy and sensitivity to trace invasive aspergillosis in hemato-oncology patients.</jats:p
Free circulating mircoRNAs support the diagnosis of invasive aspergillosis in patients with hematologic malignancies and neutropenia
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FAM13A regulates cellular senescence marker p21 and mitochondrial reactive oxygen species production in airway epithelial cells
Get PDFInhalation of noxious gasses induces oxidative stress in airway epithelial cells (AECs), which may lead to cellular senescence and contribute to the development of chronic obstructive pulmonary disease (COPD). FAM13A, a well-known COPD susceptibility gene, is highly expressed in airway epithelium. We studied whether its expression is associated with aging and cellular senescence and affects airway epithelial responses to paraquat, a cellular senescence inducer. The association between age and FAM13A expression was investigated in two datasets of human lung tissue and bronchial brushings from current/ex-smokers with/without COPD. Protein levels of FAM13A and cellular senescence marker p21 were investigated using immunohistochemistry in lung tissue from patients with COPD. In vitro, FAM13A and P21 expression was assessed using qPCR in air-liquid-interface (ALI)-differentiated AECs in absence/presence of paraquat. In addition, FAM13A was overexpressed in human bronchial epithelial 16HBE cells and the effect on P21 expression (qPCR) and mitochondrial reactive oxygen species (ROS) production (MitoSOX staining) was assessed. Lower FAM13A expression was significantly associated with increasing age in lung tissue and bronchial epithelium. In airway epithelium of patients with COPD, we found a negative correlation between FAM13A and p21 protein levels. In ALI-differentiated AECs, the paraquat-induced decrease in FAM13A expression was accompanied by increased P21 expression. In 16HBE cells, the overexpression of FAM13A significantly reduced paraquat-induced P21 expression and mitochondrial ROS production. Our data suggest that FAM13A expression decreases with aging, resulting in higher P21 expression and mitochondrial ROS production in the airway epithelium, thus facilitating cellular senescence and as such potentially contributing to accelerated lung aging in COPD.NEW & NOTEWORTHY To our knowledge, this is the first study investigating the role of the COPD susceptibility gene FAM13A in aging and cellular senescence. We found that FAM13A negatively regulates the expression of the cellular senescence marker P21 and mitochondrial ROS production in the airway epithelium. In this way, the lower expression of FAM13A observed upon aging may facilitate cellular senescence and potentially contribute to accelerated lung aging in COPD.</p
A Protective Role of FAM13A in Human Airway Epithelial Cells Upon Exposure to Cigarette Smoke Extract
No full textBackgroundChronic Obstructive Pulmonary Disease (COPD) is a progressive lung disease characterized by chronic inflammation upon inhalation of noxious particles, e.g., cigarette smoke. FAM13A is one of the genes often found to be associated with COPD, however its function in the pathophysiology of COPD is incompletely understood. We studied its role in airway epithelial barrier integrity and cigarette smoke-induced epithelial responses.Materials and MethodsProtein level and localization of FAM13A was assessed with immunohistochemistry in lung tissue from COPD patients and non-COPD controls. In vitro, FAM13A expression was determined in the absence or presence of cigarette smoke extract (CSE) in primary airway epithelial cells (AECs) from COPD patients and controls by western blotting. FAM13A was overexpressed in cell line 16HBE14o- and its effect on barrier function was monitored real-time by electrical resistance. Expression of junctional protein E-cadherin and β-catenin was assessed by western blotting. The secretion of neutrophil attractant CXCL8 upon CSE exposure was measured by ELISA.ResultsFAM13A was strongly expressed in airway epithelium, but significantly weaker in airways of COPD patients compared to non-COPD controls. In COPD-derived AECs, but not those of controls, FAM13A was significantly downregulated by CSE. 16HBE14o- cells overexpressing FAM13A built up epithelial resistance significantly more rapidly, which was accompanied by higher E-cadherin expression and reduced CSE-induced CXCL8 levels.ConclusionOur data indicate that the expression of FAM13A is lower in airway epithelium of COPD patients compared to non-COPD controls. In addition, cigarette smoking selectively downregulates airway epithelial expression of FAM13A in COPD patients. This may have important consequences for the pathophysiology of COPD, as the more rapid build-up of epithelial resistance upon FAM13A overexpression suggests improved (re)constitution of barrier function. The reduced epithelial secretion of CXCL8 upon CSE-induced damage suggests that lower FAM13A expression upon cigarette smoking may facilitate epithelial-driven neutrophilia.</jats:sec
