Development and Characterization of Experimental Autoimmune Cystitis (EAC)

Interstitial Cystitis/Painful Bladder Syndrome(IC/PBS) is a chronic inflammation of the urinary bladder, consisting of irritative voiding symptoms frequent and urgent urination and pain referred to the pelvic region or the bladder upon filling, lack of other pathology. The pathophysiology of interstitial cystitis (IC) is enigmatic. The impaired urothelium of bladder and autoimmunity might lead the underlying pathology. One of the shortcomings in IC/PBS research has been the lack of an appropriate animal model. In the current study, we show that the bladder specific Uroplakin 3A (UPK3A)-derived immunogenic UPK3A 65-84 peptide is capable of targeting experimental autoimmune cystitis (EAC) in BALB/c mice. We determined an immunogenic peptide from the known sequence of UPK3A based on having the binding motif for IAd MHC class II molecules expressed in BALB/c mice. The highly antigen-specific proliferative response to UPK3A 65-84 was determined by proliferation assays. Active immunization with the UPK3A 65-84 peptide resulted in increased micturition frequency and decreased urine output per micturition by FVC along with the increased pelvic pain response to von Frey Filaments, and decrease intercontractile intervals, bladder compliance and bladder capacity in CMG after 5 weeks of immunization compared to control group.The recall responses of LNC to UPK3A 65-84 showed selectively activated CD4+ T-cells with a proinflammatory Th1-like phenotype. Immunocytochemical analysis showed that immunization with UPK3A 65-84 peptide resulted in T-cell infiltration of the bladder. The ratio of bladder weight to body weight was increased in EAC mice simply showing bladder inflammation. The elevated gene expression levels of TNF-a, IFN-y, IL-17A, and IL-1ß were confined to the bladder but not in other organs. T-cell induction of EAC was identified by showing significantly increased micturition frequency and decreased output per micturition by FVC along with the increased pelvic pain response to von Frey monofilament stimulus a

Development and Characterization of Experimental Autoimmune Cystitis (EAC)

Interstitial Cystitis/Painful Bladder Syndrome(IC/PBS) is a chronic inflammation of the urinary bladder, consisting of irritative voiding symptoms frequent and urgent urination and pain referred to the pelvic region or the bladder upon filling, lack of other pathology. The pathophysiology of interstitial cystitis (IC) is enigmatic. The impaired urothelium of bladder and autoimmunity might lead the underlying pathology. One of the shortcomings in IC/PBS research has been the lack of an appropriate animal model. In the current study, we show that the bladder specific Uroplakin 3A (UPK3A)-derived immunogenic UPK3A 65-84 peptide is capable of targeting experimental autoimmune cystitis (EAC) in BALB/c mice. We determined an immunogenic peptide from the known sequence of UPK3A based on having the binding motif for IAd MHC class II molecules expressed in BALB/c mice. The highly antigen-specific proliferative response to UPK3A 65-84 was determined by proliferation assays. Active immunization with the UPK3A 65-84 peptide resulted in increased micturition frequency and decreased urine output per micturition by FVC along with the increased pelvic pain response to von Frey Filaments, and decrease intercontractile intervals, bladder compliance and bladder capacity in CMG after 5 weeks of immunization compared to control group.The recall responses of LNC to UPK3A 65-84 showed selectively activated CD4+ T-cells with a proinflammatory Th1-like phenotype. Immunocytochemical analysis showed that immunization with UPK3A 65-84 peptide resulted in T-cell infiltration of the bladder. The ratio of bladder weight to body weight was increased in EAC mice simply showing bladder inflammation. The elevated gene expression levels of TNF-a, IFN-y, IL-17A, and IL-1ß were confined to the bladder but not in other organs. T-cell induction of EAC was identified by showing significantly increased micturition frequency and decreased output per micturition by FVC along with the increased pelvic pain response to von Frey monofilament stimulus a

Development and Characterization of Experimental Autoimmune Cystitis (EAC)

Interstitial Cystitis/Painful Bladder Syndrome(IC/PBS) is a chronic inflammation of the urinary bladder, consisting of irritative voiding symptoms frequent and urgent urination and pain referred to the pelvic region or the bladder upon filling, lack of other pathology. The pathophysiology of interstitial cystitis (IC) is enigmatic. The impaired urothelium of bladder and autoimmunity might lead the underlying pathology. One of the shortcomings in IC/PBS research has been the lack of an appropriate animal model. In the current study, we show that the bladder specific Uroplakin 3A (UPK3A)-derived immunogenic UPK3A 65-84 peptide is capable of targeting experimental autoimmune cystitis (EAC) in BALB/c mice. We determined an immunogenic peptide from the known sequence of UPK3A based on having the binding motif for IAd MHC class II molecules expressed in BALB/c mice. The highly antigen-specific proliferative response to UPK3A 65-84 was determined by proliferation assays. Active immunization with the UPK3A 65-84 peptide resulted in increased micturition frequency and decreased urine output per micturition by FVC along with the increased pelvic pain response to von Frey Filaments, and decrease intercontractile intervals, bladder compliance and bladder capacity in CMG after 5 weeks of immunization compared to control group.The recall responses of LNC to UPK3A 65-84 showed selectively activated CD4+ T-cells with a proinflammatory Th1-like phenotype. Immunocytochemical analysis showed that immunization with UPK3A 65-84 peptide resulted in T-cell infiltration of the bladder. The ratio of bladder weight to body weight was increased in EAC mice simply showing bladder inflammation. The elevated gene expression levels of TNF-a, IFN-y, IL-17A, and IL-1ß were confined to the bladder but not in other organs. T-cell induction of EAC was identified by showing significantly increased micturition frequency and decreased output per micturition by FVC along with the increased pelvic pain response to von Frey monofilament stimulus a

Current status in cancer cell reprogramming and its clinical implications

The technology of reprogramming a terminally differentiated cell to an embryonic-like state uncovered the possibility of reprogramming a malignant cell back to a more manageable stem cell-like state. Since the current cancer models suffer from reflecting heterogeneous tumour structure and limited to express the late-stage markers, the induced pluripotent stem cell (iPSC) technology could provide an alternative model to recapitulate the early stages of cancer. Generation of iPSCs from cancer cells could offer a tool for understanding the mechanisms of tumour initiation-progression in vitro, a platform for studying tumour heterogeneity and origin of cancer stem cells and a source for cancer type-specific drug discovery studies

Myrtucommulone-A treatment decreases pluripotency- and multipotency-associated marker expression in bladder cancer cell line HTB-9

Cancer and stem cells exhibit similar features, including self-renewal, differentiation and immortality. The expression of stem-cell-related genes in cancer cells is demonstrated to be potentially correlated with cancer cell behaviour, affecting both drug response and tumor recurrence. There is an emerging body of evidence that subpopulations of tumors carry a distinct molecular sign and are selectively resistant to chemotherapy. Therefore, it is important to find novel therapeutic agents that could suppress the stem-like features of cancer cells while inhibiting their proliferation. Myrtucommulone-A (MC-A) is an active compound of a nonprenylated acylphloroglucinol isolated from the leaves of myrtle. Here we have investigated the potential of MC-A in inhibiting the expression of self-renewal regulatory factors and cancer stem cell markers in a bladder cancer cell line HTB-9. We used RT-PCR, immunocytochemistry, flow cytometry and western blotting to examine the expression of pluripotency- and multipotency-associated markers with or without treatment with MC-A. Treatment with MC-A not only decreased cancer cell viability and proliferation but also resulted in a decrease in the expression of pluripotency- and multipotency-associated markers such as NANOG, OCT-4, SOX-2, SSEA-4, TRA-1-60, CD90, CD73 and CD44. MC-A treatment was also observed to decrease the sphere-forming ability of HTB-9 cells. In summary, this study provides valuable information on the presence of stem-cell marker expression in HTB-9 cells and our results imply that MC-A could be utilized to target cancer cells with stem-like characteristics