Nanostring profiling of urothelial cells intracellularly infected with UPEC
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ABSTRACT:
PROVIDER: PRJNA780933 | ENA |
REPOSITORIES: ENA
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Project description:Urinary tract infections (UTIs) typically evoke prompt and vigorous innate immune responses in bladder, but the specific mechanisms that resolute acute inflammation remain unclear. In this study, single-cell RNA sequencing and analysis of immune cells from the bladder at early stage of uropathogenic Escherichia coli (UPEC) infection was performed. The increased hyperactivated CD137L+macrophage response and decreased immunosuppressive Treg cells are elucidated in the bladder microenvironment of UPEC infection. Conditional depletion of CD137L in macrophage aggravates inflammatory reaction, bacterial load and urothelium destruction. Deletion of Treg cells also aggravated inflammation while promoting urothelial injury after UPEC infection. Additionally, Treg cells were increased in infected bladder which can be reversed by a myeloid-CD137L deletion. We thought activation of Treg cells by CD137L+macrophage is a key mechanism for suppressing acute inflammatory reaction after UPEC infection. The present study provided new insights into the functions of myeloid-CD137L dependent Treg cells in the immune response to UPEC infection.
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Project description:Obesity is a significant public health concern associated with increased infection risk, but the mechanisms remain unclear. Using a diet induced obesity mouse model, we investigate how obesity impacts urinary tract infection (UTI) susceptibility and bladder urothelial defenses. High fat diet-fed female and male C57BL/6 mice exhibit increased susceptibility to uropathogenic E. coli (UPEC) following experimental UTI. Transcriptomic analysis of bladder urothelial cells reveals sex-specific gene expression changes, but both sexes share activation of focal adhesion and extracellular matrix signaling. Western blot and immunostaining confirm activation of focal adhesion kinase (FAK), a central component of the focal adhesion pathway, in the bladders of obese female and male mice. Mechanistically, primary human urothelial cells overexpressing FAK exhibit increased UPEC invasion. These findings demonstrate that obesity enhances UTI susceptibility and identify FAK as a conserved pathway disrupted by obesity, contributing to increased UPEC vulnerability.
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Project description:Host responses to intracellular UPEC communities; We used laser capture microdissection and microarrays to identify urothelial transcripts differentially expressed in response to proximity to intracellular UPEC. Experiment Overall Design: Transcription within four distinct populations of urothelial cells was profiled in biological duplicate: (i) uninfected, (ii) mock-infected (sterile 1x PBS), (iii) IBC-distal (cells residing >10 cell diameters from IBCs in the section plane), and (iv) IBC-proximal (cells residing <10 cell diameters from IBCs in the section plane).
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Project description:Data defines for the first time a whole bladder transcriptome of UPEC cystitis in female CBA mice using genome-wide expression profiling to map early host response pathways stemming from UPEC colonization We used microarrays to detail the global programme of gene expression in UPEC UTI in mice
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Project description:Data defines for the first time a whole bladder transcriptome of UPEC cystitis in female C57BL/6 mice using genome-wide expression profiling to map early host response pathways stemming from UPEC colonization We used microarrays to detail the global programme of gene expression in UPEC UTI in mice
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Project description:Data defines for the first time a whole bladder transcriptome of UPEC cystitis in female CBA mice using genome-wide expression profiling to map early host response pathways stemming from UPEC colonization We used microarrays to detail the global programme of gene expression in UPEC UTI in mice Whole mouse bladder collected at 2h following transurethral infection for RNA extraction and hybridization on Affymetrix microarrays. Non-pooled samples were used, with one array per mouse to expand power of expression profiles. Control mice received only PBS as a mock infection. Group sizes were five mice per group, and therefore five arrays were used per group.
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Project description:Data defines for the first time a whole bladder transcriptome of UPEC cystitis in female C57BL/6 mice using genome-wide expression profiling to map early host response pathways stemming from UPEC colonization We used microarrays to detail the global programme of gene expression in UPEC UTI in mice Whole mouse bladder collected at 2h following transurethral infection for RNA extraction and hybridization on Affymetrix microarrays. Non-pooled samples were used, with one array per mouse to expand power of expression profiles. Control mice received only PBS as a mock infection. Group sizes were five mice per group, and therefore five arrays were used per group.
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