ABSTRACT: Transcription profiling of lung from a mouse model of cystic fibrosis fromB6 and Balb strains vs wild type littermates to determine strain specific differences
Project description:The murine pulmonary response is strain specific. Susceptible strains such as C57BL6/J experience an inflammatory response followed by progressive lung disease. Resistant strains such as Balb/c do not experience significant degrees of inflammation or fibrosis. This study aims to determine the genetic basis of sensitivity differences between strains Keywords: other
Project description:Cystic fibrosis (CF) intestinal disease is characterized by alterations in processes such as proliferation and apoptosis which are known to be regulated in part by microRNA’s. Herein, we completed microRNA expression profiling of the intestinal tissue from the cystic fibrosis mouse model of cystic fibrosis transmembrane conductance regulator (Cftr) deficient mice (BALBc/J Cftrtm1UNC), relative to that of wildtype littermates, to determine whether changes in microRNA expression level are part of this phenotype. We identified 24 microRNA's to be significantly differentially expressed in tissue from CF mice compared to wildtype, with the higher expression in tissue from CF mice. These data were confirmed with real time PCR measurements. A comparison of the list of genes previously reported to have decreased expression in the BALB x C57BL/6J F2 CF intestine to that of genes putatively targeted by the 24 microRNA’s, determined from target prediction software, revealed 20% of the gene expression profile to overlap with predicted targets. Pathway analysis identified these common genes to function in phosphatase and tensin homolog-, protein kinase A-, phosphoinositide-3 kinase/Akt- and peroxisome proliferator-activated receptor alpha/retinoid X receptor alpha signaling pathways, among others, and through real time PCR experiments genes of these pathways were demonstrated to have lower expression in the BALB CF intestine. We conclude that altered microRNA expression is a feature which putatively influences both metabolic abnormalities and the altered tissue homeostasis component of CF intestinal disease. Two condition experiment, Balbc/J Cftrtm1UNC -/- (Cystic Fibrosis (CF) Mice) and Balbc/J Cftrtm1UNC +/+ (Wild Type (WT) Mice). Biological Replicates: 7 WT, 8CF. Ileum Tissue.
Project description:The murine pulmonary response is strain specific. Susceptible strains such as C57BL6/J experience an inflammatory response followed by progressive lung disease. Resistant strains such as Balb/c do not experience significant degrees of inflammation or fibrosis. This study aims to determine the genetic basis of sensitivity differences between strains
Project description:Cystic fibrosis (CF) intestinal disease is characterized by alterations in processes such as proliferation and apoptosis which are known to be regulated in part by microRNA’s. Herein, we completed microRNA expression profiling of the intestinal tissue from the cystic fibrosis mouse model of cystic fibrosis transmembrane conductance regulator (Cftr) deficient mice (BALBc/J Cftrtm1UNC), relative to that of wildtype littermates, to determine whether changes in microRNA expression level are part of this phenotype. We identified 24 microRNA's to be significantly differentially expressed in tissue from CF mice compared to wildtype, with the higher expression in tissue from CF mice. These data were confirmed with real time PCR measurements. A comparison of the list of genes previously reported to have decreased expression in the BALB x C57BL/6J F2 CF intestine to that of genes putatively targeted by the 24 microRNA’s, determined from target prediction software, revealed 20% of the gene expression profile to overlap with predicted targets. Pathway analysis identified these common genes to function in phosphatase and tensin homolog-, protein kinase A-, phosphoinositide-3 kinase/Akt- and peroxisome proliferator-activated receptor alpha/retinoid X receptor alpha signaling pathways, among others, and through real time PCR experiments genes of these pathways were demonstrated to have lower expression in the BALB CF intestine. We conclude that altered microRNA expression is a feature which putatively influences both metabolic abnormalities and the altered tissue homeostasis component of CF intestinal disease.
Project description:The opportunistic pathogen Pseudomonas aeruginosa is among the main colonizers of the lungs of cystic fibrosis (CF) patients. We have isolated and sequenced several P. aeruginosa isolates from the sputum of CF patients and used phenotypic, genomic and proteomic analyses to compare these CF derived strains with each other and with the model strain PAO1.
Project description:To determine if differences in the severity of pulmonary infection in cystic fibrosis been seen in late isolates od Pseudomonas aeruginosa and Burkholderia cepacia are associated with differences in the initial repsonse of alveolar macrophages (AM) to these pathogens, we assessed gene expression changes in human AM in response to infection with a laboratoty strain, early and late clinical isolates of P. aeruginosa, and B. cepacia. Experiment Overall Design: Alveolar macrophages were obtained from bronchoalveolar lavage. Experiment Overall Design: Two clinical strains isolated from the sputum of an individual with CF, AD2A and AD15B (provided by J. Burns, University of Washington, Seattle). AD2A is an early clinical isolate, and AD15B is a late clinical isolate; both were derived from the same individual.
Project description:Rationale: Chagasic disease is associated with high morbidity in Latin America. Acute Chagasic myocarditis is consistently found in acute infections but little is known about its contribution to chronic cardiomyopathy. The aim of the study was to phenotypically characterize two strains of mice with differential Chagas infection susceptibility and correlate strain myocarditis phenotypes with heart tissue gene expression. Methods: C57BL/6J and Balb/c mice were injected intraperitoneally with 0 or 150-200 tissue-derived trypomastigotes (Tulahuen strain). Echocardiograms, brain natriuretic peptide and troponin were measured. Heart tissue was harvested for histopathological analysis and gene expression profiling on microarrays. Genes differently expressed between infected Balb/c and C57BL/6J were identified Results: Echocardiograms demonstrated differences in heart rate in Balb/c vs. C57BL/6J infected mice: 413 vs. 476 bpm, (p=0.0001), stroke volume: 31.9 ± 9.3 vs. 39.2 ± 5.5 µl (p=0.03); and cardiac output: 13.1 ± 3.5 vs. 18.7 ± 3.2 µl/min (p=0.002). Gene expression at 4 weeks analysis demonstrated 32 statistically significant (q-value < 0.05) differentially expressed genes between infected Balb/c and C57BL/6J which were enriched for genes related to protein kinase B (AKT) pathway
Project description:To determine if differences in the severity of pulmonary infection in cystic fibrosis been seen in late isolates od Pseudomonas aeruginosa and Burkholderia cepacia are associated with differences in the initial repsonse of alveolar macrophages (AM) to these pathogens, we assessed gene expression changes in human AM in response to infection with a laboratoty strain, early and late clinical isolates of P. aeruginosa, and B. cepacia. Keywords: Comparison of gene expression in alveolar macrophages of normal non-smokers and normal smokers.
Project description:<p>While bacterial metabolism is known to impact antibiotic efficacy and virulence, the metabolic capacities of individual microbes in cystic fibrosis lung infections are difficult to disentangle from sputum samples. Here, we show that untargeted metabolomic profiling of supernatants of multiple strains of<em> Pseudomonas aeruginosa</em> and <em>Staphylococcus aureus </em>grown in monoculture in synthetic cystic fibrosis media (SCFM) reveal distinct species-specific metabolic signatures with limited strain-to-strain variability. The majority of metabolites significantly consumed by <em>S. aureus </em>were also consumed by <em>P. aeruginosa</em>, indicating that <em>P. aeruginosa</em> has the flexibility to metabolically outcompete<em> S. aureus </em>in coculture even in the absence of other pathogen-pathogen interactions. Finally, metabolites that were uniquely produced by one species or the other were identified. Specifically, the virulence factor precursor anthranilic acid as well as the quinoline 2,4-Quinolinediol (DHQ) were robustly produced across all tested strains of <em>P. aeruginosa</em>. Through the direct comparison of the extracellular metabolism of <em>P. aeruginosa</em> and <em>S. aureus</em> in a physiologically relevant environment, this work provides insight towards the potential metabolic interactions in vivo and supports the development of species-specific diagnostic markers of infection.</p>
Project description:Distinct genotypic and pathogenicity differences exist between strains of the opportunistic protozoan Toxoplasma gondii. The label-free quantitative acetylomics approach was applied to investigate the differences in the level of acetylation between RH strain, PRU strain and PYS strain of T. gondii. The results showed that lysine acetylation in RH strain was the largest (458 acetylated proteins) among the three strains. The PYS strain had the second-largest acetylome (188 acetylated proteins), whereas lysine acetylation in PRU strain (115 acetylated proteins) was the smallest. Motif analysis revealed four, three and four motifs enriched from lysine acetylation sequences in RH strain, PRU strain and PYS strain respectively, suggesting specificity of acetyltransferases in these strains. Our analysis also identified 15 DAPs (differentially expressed acetylated proteins), 3 DAPs and 26 DAPs in RH strain vs PRU strain, PRU strain vs PYS strain and PYS strain vs RH strain, respectively. Compared with PYS strain and PRU strain, histone acetyltransferase and glycyl-tRNA synthase had higher expression levels in RH strain, reflecting genotype-specific differences in stress tolerance. These findings provide novel insight into the acetylomic profiles of major genotypes of T. gondii and provide an important resource for further analysis of the roles of the acetylated parasite proteins in the regulation of cellular functions.