Project description:Purpose: The goals of this study are to compare 1. The transcription profile in KDM6A wildtype and KDM6A mutated urothelial bladder carcinoma. 2. The transcriptional changes in KDM6A mutated urothelial bladder carcinoma upon EZH2 inhibitor treatment.
Project description:Hunner-type interstitial cystitis/bladder pain syndrome (HIC) is a complex and clinically challenging chronic disorder characterized by bladder-centered symptoms, including persistent pelvic pain and urinary urgency frequency, with elusive pathogenesis involving poorly defined multi-system interactions and systemic implications. To investigate upstream drivers of urothelial injury, we employed an integrative multi-omics approach, incorporating metagenomic sequencing, targeted metabolomics of urine and serum, and single-cell RNA sequencing. Our analysis identified Enterococcus avium, Megasphaera micronuciformis, and Fusobacterium nucleatum as the significantly enriched species in HIC patients, with bile acid metabolism appearing as a prominently altered pathway. Metabolomic profiling revealed the elevated levels of taurochenodeoxycholic acid (TCDCA) and tauroursodeoxycholic acid, while single-cell transcriptomic data pointed to TLR3 signaling as a key mediator of urothelial injury. Cross-omics correlation proposed a mechanistic axis involving E. avium, TCDCA, and TLR3 activation. Functional validation using fecal microbiota transplantation, E. avium transplantation, and TCDCA administration demonstrated that this axis contributes to the urothelial barrier disruption and inflammatory responses via TLR3. Intervention with a TLR3 inhibitor demonstrated therapeutic potential. These findings shed light on the pathogenesis of HIC, highlighting the potential role of the gut-bladder axis in disease development. This may contribute not only to the discovery of novel biomarkers but also to the development of personalized treatment strategies targeting specific molecular and microbial pathways.
Project description:Characterization of the gene expression profiles of specific cell populations of the human urinary bladder provides an important set of research tools for the study of cellular differentiation and the cancer process. The transcriptome is a definitive identifier of each individual cell types. Surgically resected tissue was digested by collagenase and the different cell types were sorted by antibodies to cluster designation (CD) cell surface antigens. The sorted cells were analyzed by DNA microarrays. The transcriptome datasets were analyzed for differentially expressed genes and plotted on a principal components analysis space for cell lineage relationship. The following bladder cell types were analyzed: CD9+ urothelial, CD104+ basal, CD13+ stromal of lamina propria, CD9+ urothelial carcinoma cancer, and CD13+ urothelial carcinoma-associated stromal. Gene expression differences between the cell types of tumor and their respective non-cancer counterpart provide biomarker candidates. Basal cells of the bladder and prostate, although sharing CD cell surface markers, are quite different in overall gene expression. Furthermore, these cells lack transcript features of stem cell signature of embryonic stem or embryonal carcinoma cells. Cell type-specific transcriptomes are more informative than bulk tissue transcriptomes. The relatedness of different cell types can be determined by transcriptome dataset comparison. Bladder cell types were sorted from tissue specimens, and analyzed by DNA microarrays. The various transcriptomes were compared by principal components analysis for cell lineage relationship.
Project description:<p>Hunner-type interstitial cystitis/bladder pain syndrome (HIC) is a complex and clinically challenging chronic disorder characterized by bladder-centered symptoms, including persistent pelvic pain and urinary urgency frequency, with elusive pathogenesis involving poorly defined multi-system interactions and systemic implications. To investigate upstream drivers of urothelial injury, we employed an integrative multi-omics approach, incorporating metagenomic sequencing, targeted metabolomics of urine and serum, and single-cell RNA sequencing. Our analysis identified Enterococcus avium, Megasphaera micronuciformis, and Fusobacterium nucleatum as the significantly enriched species in HIC patients, with bile acid metabolism appearing as a prominently altered pathway. Metabolomic profiling revealed the elevated levels of taurochenodeoxycholic acid (TCDCA) and tauroursodeoxycholic acid, while single-cell transcriptomic data pointed to TLR3 signaling as a key mediator of urothelial injury. Cross-omics correlation proposed a mechanistic axis involving E. avium, TCDCA, and TLR3 activation. Functional validation using fecal microbiota transplantation, E. avium transplantation, and TCDCA administration demonstrated that this axis contributes to the urothelial barrier disruption and inflammatory responses via TLR3. Intervention with a TLR3 inhibitor demonstrated therapeutic potential. These findings shed light on the pathogenesis of HIC, highlighting the potential role of the gut-bladder axis in disease development. This may contribute not only to the discovery of novel biomarkers but also to the development of personalized treatment strategies targeting specific molecular and microbial pathways.</p>
Project description:mRNAseq data for differentiated and mitotically-quiescent Normal Human Urothelial (NHU) cells at 14 days post infection (dpi) with BKPyV