Project description:The primary goals of this study are to: determine how intracellular infection of urothelial cells with uropathogenic Escherichia coli influences urothelial cell metabolism, and determine the influence of cytochrome bd on the urothelial cell response to infection
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.
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.
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).
Project description:The NLRP3 inflammasome, estrogen and antimicrobial peptides have all been emphasised to have a vital role in the protection of the bladder urothelium. However, the interdependence between these protective factors during a bladder infection is currently unknown. Our aim was to investigate the role of NLRP3 in regulation of antimicrobial peptides and estrogen signaling in bladder epithelial cells during a UPEC infection. Human bladder epithelial cells and CRISPR/Cas9 generated NLRP3-deficient cells were stimulated with the UPEC strain CFT073 and estradiol. The gene and protein expression were evaluated with microarray, qRT-PCR, western blot and ELISA. Microarray results showed that the expression of most antimicrobial peptides was reduced in CFT073-infected NLRP3-deficient cells compared to Cas9 control cells. Conditioned medium from NLRP3-deficient cells also lost the ability to suppress CFT073 growth. Moreover, NLRP3-deficient cells had lower basal release of Beta-defensin-1, Beta-defensin-2 and RNase7. The ability of estradiol to induce an increased expression of antimicrobial peptides was also abrogated in NLRP3-deficient cells. The decreased antimicrobial peptide expression might be linked to the observed reduced expression and activity of estradiol receptor beta in NLRP3-deficient cells. This study suggests that NLRP3 may regulate the release and expression of antimicrobial peptides and affect estrogen signaling in bladder epithelial cells.
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
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
Project description:In this study, we aimed to determine the overall effect of TNFα on transcriptomic profile of urothelial cells using RNA sequencing. Our study represents a reference database that could lead to a better understanding of the role of urothelial cells in IC/BPS pathology and more importantly, sets grounds for future studies exploring potential biomarkers and therapeutic targets for IC/PBS diagnosis and treatment.
Project description:Expression profiling of a panel of urothelial cancer cells. The goal of the study is to exam the genome wide expression profile in each of the 30 urothelial cancer cells tested in our laboratory
Project description:This study used the NanoString nCounter hybridization system and nCounter miRNA expression assays to identify and quantitate circulating cellular miRNAs during HIV-1 elite suppression, active HIV-1 replication, and uninfected status. Blood samples were from eight uninfected controls, seven HIV-1 elite suppressors with undetectable viral load, and six viremic HIV-1-infected patients.