Project description:Bacteroidales strain CF is the most abundant nondechlorinating organism in a Dehalobacter-containing enrichment culture that consistently reductively dechlorinates >50 mg/liter chloroform or 1,1,1-trichloroethane (methyl chloroform). We assembled and closed the complete genome sequence of this organism from the metagenomic sequencing data for enrichment cultures. This organism is predicted to ferment l-lactate and ethanol.
Project description:We did bulk RNA sequencing in newborn cystic fibrosis (CF) and non-CF pig kidney. We compared kidney gene expression profiling between non-CF and CF pigs. RNA sequencing results showed that there is not significant difference between non-CF and CF in terms of gene expression, suggesting that CFTR knockout does not affect kidney development in newborn pigs.
Project description:A small-scale whole genome microarray study of gene expression in human native nasal epithelial cells from F508del-CFTR homozygous CF patients and non-CF controls. We used the custom designed Affymetrix HsAirwaya520108F Arrays to compare gene expression in 5 CF and 5 non CF nasal epithelial cell samples. We analysed a total of 10 samples (5 CF and 5 non CF). The CF group contained 2 males and 3 females, with an average age of 14 years and an average of 6% inflammatory cells per sample, and the non CF group contained 3 males and 2 females with an average age of 14.8 years and an average of 4.7% inflammatory cells.
Project description:A small-scale whole genome microarray study of gene expression in human native nasal epithelial cells from F508del-CFTR homozygous CF patients and non-CF controls. We used the custom designed Affymetrix HsAirwaya520108F Arrays to compare gene expression in 5 CF and 5 non CF nasal epithelial cell samples.
Project description:Mutations in CFTR have been shown to alter the immune response of macrophages, for example, by reducing the ability of macrophages to phagocytose and kill bacteria. This contributes to chronic bacterial infection and inflammation in the lungs, which leads to significant morbidity and mortality in cystic fibrosis (CF). Extracellular vesicles (EVs) are secreted by a variety of cell types in the lungs and participate in the host immune response to bacterial infection. However, nothing is known about the effect of EVs secreted by CF airway epithelial cells (AEC) on CF macrophages. Therefore, we examined the effect of EVs secreted by primary CF AEC on CF monocyte derived macrophages (MDM) and compared it with the effect of EVs secreted by wild type (WT) AEC on WT MDM. EVs increased pro-inflammatory cytokine secretion and enhanced the expression of numerous innate immune genes in WT MDM. However, the response of CF MDM to EVs was significantly attenuated compared to WT MDM, a difference that was also observed when EVs were isolated from WT and CF AEC exposed to Pseudomonas aeruginosa. Attenuated responses by CF MDM can be attributed to defects in the CF macrophages themselves rather than differences between CF and WT EVs, because EVs secreted by CF AEC or WT AEC elicited similar cytokine secretion by CF MDM. EVs secreted by P. aeruginosa exposed AEC resulted in the upregulation of immune response genes and increased secretion of pro-inflammatory cytokines, chemoattractants and chemokines involved in tissue repair by WT MDM, whereas the response of CF MDM was attenuated by comparison. To our knowledge, this is the first study examining the effect of EVs secreted by CF AEC on CF MDM, and it demonstrates that the Phe508del mutation in CFTR attenuates the innate immune response of MDM to EVs.
Project description:Cystic Fibrosis (CF) is associated with pathology in multiple tissues including the lung, digestive tract and reproductive system. Lung disease is primarily a post-natal event but other organs are affected before birth. Here we use the CF sheep model to investigate the initiation and progression of CF disease through gestation.
Project description:The goal of this study was to compare cell composition and gene expression patterns for different cell types in large and small airways of CFTR+/+ (non-CF) and CFTR-/- (CF) pigs.
Project description:Cystic fibrosis (CF), a genetic disorder, is characterized by chronic lung disease. Small non-coding RNAs are key regulators of gene expression and participate in various processes, which are dysregulated in CF; however, they remain poorly studied. Here, we determined the complete microRNAs (miRNAs) expression pattern in three CF ex-vivo models. The miRNA profiles of air-liquid interface cultures of airway epithelia (bronchi, nasal cells, and nasal polyps) samples from patients with CF and non-CF controls were obtained by deep sequencing. Compared with non-CF controls, several miRNAs were deregulated in CF samples, for instance miR-181a-5p and the miR-449 family were upregulated. Moreover, mature miRNAs often showed variations (i.e., isomiRs) relative to their reference sequence, such as miR-101, suggesting that miRNAs consist of heterogeneous repertoires of multiple isoforms with different effects on gene expression. Analysis of miR-181a-5p and miR-101-3p roles indicated that they regulate the expression of WISP1, a key component of cell proliferation/migration programs. We showed that miR-101 and miR-181a-5p participated in aberrant recapitulation of wound healing programs by controlling WISP1 mRNA and protein level. Our miRNA expression data bring new insights into CF physiopathology and define new potential therapeutic targets in CF
Project description:Response of A549 cells treated with Aspergillus fumigatus wild type culture filtrate (WT-CF) or PrtT protease deficient mutant culture filtrate (PrtT-CF) for 8h Aspergillus fumigatus is the most commonly encountered mold pathogen of humans, predominantly infecting the respiratory system. Colonization and penetration of the lung alveolar epithelium is a key but poorly understood step in the infection process. This study focused on identifying the transcriptional and cell-signaling responses activated in A549 alveolar carcinoma cells incubated in the presence of A. fumigatus wild-type and ΔPrtT protease-deficient germinating conidia and culture filtrates (CF). Microarray analysis of exposed A549 cells identified distinct classes of genes whose expression is altered in the presence of germinating conidia and CF and suggested the involvement of both NFkB and MAPK signaling pathways in mediating the cellular response. Phosphoprotein analysis of A549 cells confirmed that JNK and ERK1/2 are phosphorylated in response to CF treatment in a protease-dependent manner. Inhibition of JNK or ERK1/2 kinase activity substantially decreased CF-induced cell damage, including cell peeling, actin-cytoskeleton damage, and reduction in metabolic activity and necrotic death. These results suggest that inhibition of MAPK-mediated host responses to treatment with A. fumigatus CF decreases cellular damage, a finding with possible clinical implications.