Project description:These experiments are designed to profile of how TcdB and XMU-MP-1 modify gene expression in pericytes. TcdB is a bacterial toxin and primary virulence factor produced by the enteric pathogen C. difficile, and pericytes are putative target cells for TcdB in the colon. XMU-MP-1 is an inhibitor of a core kinase (MST1/2) in the Hippos signaling pathway, and TcdB activates Hippo signaling, which is possibly critical for the pathogenesis of C. difficile disease. RNA-seq was carried out and the resulting gene expression profiles were analyzed focusing on CSPG4 (TcdB receptor) and a panel of genes that are regulated by YAP and TAZ (primary transcriptional regulator in Hippo pathway). We also focused on genes regulated by MTF1, which is another transcription regulator that can be controlled by Hippo signaling.
Project description:The Clostridioides difficile toxins TcdA and TcdB are responsible for diarrhea and colitis. The aim of this project was to explore the effects of the toxins on epithelial barrier function and the molecular mechanisms for diarrhea and inflammation. RNA-seq of toxin-treated intestinal cell monolayers was performed to describe the C. difficile-mediated effects. mRNA profiles from intestinale epithelial cells were generated by deep sequencing using Illumina NovaSeq 6000. This data provide the basis for subsequent upstream regulator analysis.
Project description:Gene expression level of Clostridioides difficile (C. difficile) strain R20291 comparing control C. difficile carring pMTL84151 as vector plasmid with C. difficile conjugated with a pMTL84151-03890 gene. Goal was to determine the effects of 03890 gene conjugation on C. difficile strain R20291 gene expression.
Project description:This experiment uses a CUT&RUN approach to determine if the CCCTC-binding factor (CTCF) is altered in its ability to bind the human genome following exposure to TcdB. TcdB is a bacterial toxin and primary virulence factor produced by the enteric pathogen C. difficile. CTCF is a DNA binding protein that regulates gene transcription in the host.
Project description:Clostridium difficile is an important nosocomial pathogen and the leading cause of hospital-acquired diarrhea. Antibiotic use is the primary risk factor for the development of C. difficile-associated disease because it disrupts normal protective gut flora and enables C. difficile to colonize the colon. C. difficile damages host tissue by secreting toxins and disseminates by forming spores. The toxin-encoding genes, tcdA and tcdB are part of a pathogenicity locus, which also encodes the gene tcdR that codes for the toxin genes positive regulator. TcdR is an alternate sigma factor that initiates transcription of tcdA and tcdB at their promoters. Alternative sigma factors are known to regulate virulence and virulence associated genes in many pathogenic bacteria. We created a tcdR mutant in the epidemic-type C. difficile R20291 strain in an attempt to identify the global role of tcdR. A site-directed mutation in tcdR affected both toxin production and sporulation in C. difficile R20291. Spores derived from the tcdR mutant were found to be mildly temperature sensitive. Moreover, nearly two fold more taurocholate was needed to germinate spores from the tcdR mutant than the spores prepared from the wild-type parent strain. Comparison of the tcdR mutant transcriptome with the parent strain revealed many differentially expressed late sporulation genes in the tcdR mutant. These data suggests that gene regulatory networks of toxin production and sporulation in Clostridium difficile are linked with each other.