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Expression data profile of A498 cells treated with DRB or E. coli 83972

ABSTRACT: Urinary tract infections (UTIs) constitute a highly relevant model of microbial adaptation, in which the contrasting effects of pathogens and commensals on host tissues are clearly displayed. While virulent Escherichia coli cause severe, potentially life-threatening disease by breaking the inertia of the mucosal barrier and infecting the kidneys, the most common outcome of bacteriuria is an asymptomatic carrier state resembling commensalism at other mucosal sites. It remains unclear if the lack of destructive inflammation merely reflects low virulence or if carrier strains actively inhibit disease associated responses in the host. To address this question, we examined the effects of asymptomatic bacterial carriage on host gene expression. The asymptomatic strain E. coli 83972 caused reduction in Pol II phosphorylation in the nuclei of human kidney epithelial A498 cells. To specifically address if Pol II inhibition alters the response to infection, A498 cells were pretreated with 5,6-dichloro-1-b-D-ribofuranosylbenzimidazole (DRB). This adenosine analogue has been proposed to specifically and reversibly inhibit Pol II transcription without directly affecting other cellular functions. A498 cultered cells were infected with E. coli 83972 or DRB for 4 hours. The culture medium with DMSO was used as a background control.

ORGANISM(S): Homo sapiens

PROVIDER: GSE43886 | GEO | 2013/02/23

SECONDARY ACCESSION(S): PRJNA190084

REPOSITORIES: GEO

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Expression data profile of A498 cells treated with DRB or E. coli 83972

Project description:Urinary tract infections (UTIs) constitute a highly relevant model of microbial adaptation, in which the contrasting effects of pathogens and commensals on host tissues are clearly displayed. While virulent Escherichia coli cause severe, potentially life-threatening disease by breaking the inertia of the mucosal barrier and infecting the kidneys, the most common outcome of bacteriuria is an asymptomatic carrier state resembling commensalism at other mucosal sites. It remains unclear if the lack of destructive inflammation merely reflects low virulence or if carrier strains actively inhibit disease associated responses in the host. To address this question, we examined the effects of asymptomatic bacterial carriage on host gene expression. The asymptomatic strain E. coli 83972 caused reduction in Pol II phosphorylation in the nuclei of human kidney epithelial A498 cells. To specifically address if Pol II inhibition alters the response to infection, A498 cells were pretreated with 5,6-dichloro-1-b-D-ribofuranosylbenzimidazole (DRB). This adenosine analogue has been proposed to specifically and reversibly inhibit Pol II transcription without directly affecting other cellular functions. A498 cultered cells were infected with E. coli 83972 or DRB for 4 hours. The culture medium with DMSO was used as a background control. A498 cells were infected with E. coli 83972 or DRB for 4 h. Isolated RNA was subjected to whole genome transcriptome analysis.

2013-02-23 | E-GEOD-43886 | biostudies-arrayexpress

Expression data profile of A498 cells inoculated with E. coli 83972 and E. coli CFT073

Project description:Urinary tract infections (UTIs) constitute a highly relevant model of microbial adaptation, in which the contrasting effects of pathogens and commensals on host tissues are clearly displayed. While virulent Escherichia coli cause severe, potentially life-threatening disease by breaking the inertia of the mucosal barrier and infecting the kidneys, the most common outcome of bacteriuria is an asymptomatic carrier state resembling commensalism at other mucosal sites. It remains unclear if the lack of destructive inflammation merely reflects low virulence or if carrier strains actively inhibit disease associated responses in the host. To address this question, we examined the effects of asymptomatic bacterial carriage on host gene expression. A498 cell line has been validated as a model of uropathogenic E. coli infection; the cells express functional receptors for bacterial virulence ligands and the response to virulent strains reflects human UTI. The cells were infected with asymptomatic and pathogenic E. coli in vitro, and harvested RNA was subjected to whole genome transcriptome analysis. A498 human kidney epithelial cells were infected with the asymptomatic (E. coli 83972) or virulent strains (E. coli CFT073) for 4 hours. The cells with culture medium alone were used as a control. The experiment was performed in biological duplicates or triplicates.

2013-02-23 | E-GEOD-43790 | biostudies-arrayexpress

Expression data profile of A498 cells inoculated with E. coli 83972 and E. coli CFT073

Project description:Urinary tract infections (UTIs) constitute a highly relevant model of microbial adaptation, in which the contrasting effects of pathogens and commensals on host tissues are clearly displayed. While virulent Escherichia coli cause severe, potentially life-threatening disease by breaking the inertia of the mucosal barrier and infecting the kidneys, the most common outcome of bacteriuria is an asymptomatic carrier state resembling commensalism at other mucosal sites. It remains unclear if the lack of destructive inflammation merely reflects low virulence or if carrier strains actively inhibit disease associated responses in the host. To address this question, we examined the effects of asymptomatic bacterial carriage on host gene expression. A498 cell line has been validated as a model of uropathogenic E. coli infection; the cells express functional receptors for bacterial virulence ligands and the response to virulent strains reflects human UTI. The cells were infected with asymptomatic and pathogenic E. coli in vitro, and harvested RNA was subjected to whole genome transcriptome analysis.

2013-02-23 | GSE43790 | GEO

Gene expression profile in patients inoculated with E. coli 83972

Project description:Urinary tract infections (UTIs) constitute a highly relevant model of microbial adaptation, in which the contrasting effects of pathogens and commensals on host tissues are clearly displayed. While virulent Escherichia coli cause severe, potentially life-threatening disease by breaking the inertia of the mucosal barrier and infecting the kidneys, the most common outcome of bacteriuria is an asymptomatic carrier state resembling commensalism at other mucosal sites. It remains unclear if the lack of destructive inflammation merely reflects low virulence or if carrier strains actively inhibit disease associated responses in the host. To address this question, we examined the effects of asymptomatic bacterial carriage on host gene expression. Therapeutic urinary tract inoculation with the prototype ABU strain E. coli 83972 is a safe alternative approach in patients with therapy-resistant recurrent UTI. The strain establishes persistent bacteriuria, protecting patients against super-infection with more virulent strains. Using this protocol, we examined if the establishment of asymptomatic bacterial carriage alters host gene expression.

2013-02-23 | GSE43838 | GEO

Gene expression profile in patients inoculated with E. coli 83972

Project description:Urinary tract infections (UTIs) constitute a highly relevant model of microbial adaptation, in which the contrasting effects of pathogens and commensals on host tissues are clearly displayed. While virulent Escherichia coli cause severe, potentially life-threatening disease by breaking the inertia of the mucosal barrier and infecting the kidneys, the most common outcome of bacteriuria is an asymptomatic carrier state resembling commensalism at other mucosal sites. It remains unclear if the lack of destructive inflammation merely reflects low virulence or if carrier strains actively inhibit disease associated responses in the host. To address this question, we examined the effects of asymptomatic bacterial carriage on host gene expression. Therapeutic urinary tract inoculation with the prototype ABU strain E. coli 83972 is a safe alternative approach in patients with therapy-resistant recurrent UTI. The strain establishes persistent bacteriuria, protecting patients against super-infection with more virulent strains. Using this protocol, we examined if the establishment of asymptomatic bacterial carriage alters host gene expression. After antibiotic treatment to remove prior infection, patients were inoculated with E. coli 83972 through a catheter. Blood samples were obtained before and 24 h after inoculation.

2013-02-23 | E-GEOD-43838 | biostudies-arrayexpress

Expression data profile of A498 cells infected with E. coli 83972, mutants or stimualted with rNlpD

Project description:Unlike pathogens, which attack the host, commensal bacteria create a state of friendly coexistence. Here, we identify a new mechanism of bacterial adaptation to the host niche, where they reside. Asymptomatic carrier strains are shown to inhibit RNA Polymerase II (Pol II) in host cells by targeting Ser2 phosphorylation; a step required for productive mRNA elongation. Assisted by a rare, spontaneous loss-of-function mutant from a human carrier, the bacterial NlpD protein is identified as a Pol II inhibitor. After internalization by host cells, NlpD is shown to target constituents of the Pol II phosphorylation complex (RPB1 and PAF1C), attenuating host gene expression. Therapeutic efficacy of the rNlpD protein is demonstrated in a urinary tract infection model, by reduced tissue pathology, accelerated bacterial clearance and attenuated Pol II-dependent gene expression. The findings suggest an intriguing, evolutionarily conserved mechanism for bacterial modulation of host gene expression, with a significant therapeutic potential.

2020-12-11 | GSE162190 | GEO

MicroRNAs regulate differential innate immune response to uropathogenic and commensal-like Escherichia coli infection in the surrogate insect model Galleria mellonella

Project description:Identification and expression analysis of microRNAs in infected larvae of the insect model Galleria mellonella with uropathogenic (UPEC) and commensal E. coli strains that are known to cause symptomatic and asymptomatic bacteriuria (ABU) in humans, respectively.

2018-12-18 | GSE123965 | GEO

Host imprinting on bacterial genomes-rapid, divergent evolution in individual hosts

Project description:Transcriptional profiling of Asymptomatic Bacteriuria (ABU) Escherichia coli strain 83972 comparing the progenitor wild type strain ABU83972 with its re-isolates from human bladder colonization (PI-2, PII-4, PIII-4) and in vitro cultivation experiment (4.9).

2011-01-15 | GSE19917 | GEO

DRB-PRO-Seq for measuring Pol II transcriptional elongation rate alterations upon inhibition of analog sensitive CDK12 and/or CDK13

Project description:We performed PRO-Seq to determine Pol II elongation rates by measruing the distance of Pol II travels during 25 minutes after Pol II being relased from DRB induced pausing. We allowed Pol II to release in the presence of 1-NM-PP1 or DMSO after DRB being wahsed out in order to assess the effects of CDK12 and/or CDK13 inhibtion on transcriptional elongation rates.

2020-04-29 | GSE141373 | GEO

DRB-PRO-Seq for measuring Pol II transcriptional elongation rate alteration up on CDK12 and CDK13 inhibition by THZ531

Project description:We performed PRO-Seq to measure the Pol II elongation rate by measruing the distance of Pol II travels at viarous time points after relased from DRB induced pausing. We compared DMSO/DRB or THZ531/DRB co-treated sample to assess the effect of CDK12 and CDK13 inhibtion by THZ531 on transcriptional elongation rate.

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