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• W4377029208 abstract "<b>Abstract ID 15021</b> <b>Poster Board 127</b> Bladder inflammation, overactivity, and pain are adverse side effects of Interstitial Cystitis/Bladder Pain Syndrome (IC/BPS), a pathological bladder condition characterized by urinary urgency, bladder pain, and inflammation that affects up to 17% of American women. IC/BPS has proven to be resistant to standard anti-inflammatory treatments and analgesics, therefore research has focused on developing new treatments for IC/BPS-associated chronic pain. One such potential avenue for treatment lies in the expression of non-native, engineered ion channels in bladder afferent nerves that can selectively reduce nerve excitability in response to inflammatory conditions or after activation with a designer ligand. To this end, we have created an engineered chloride channel, EG3RF, that is activated by both the acidic pH that is a hallmark of inflammation as well as bioactive amines, such as cysteamine or propylamine. We have previously shown that expression/activation of this channel in the bladder can inhibit bladder hyperactivity in cyclophosphamide (CYP) treated rats (a model of IC/BPS), likely by hyperpolarizing bladder afferent nerves. Surprisingly, preliminary data suggested that EG3RF activity also reduced CYP-induced bladder inflammation, suggesting secondary actions on the urothelium itself. Bladder inflammation occurs downstream of urothelial ATP release by lysosomal exocytosis, which is associated with increased lysosomal calcium release. Thus, we hypothesized that EG3RF activation may inhibit bladder inflammation by interfering with this signaling in urothelial cells. Fura-2 based calcium imaging demonstrated that EG3RF activation in urothelial cells decreases the calcium response to the TRPML1 agonist ML-SA1. Furthermore, while expression of EG3RF alone slightly decreases calcium release (-22.15%), cotreatment of EG3RF-transfected cells with ML-SA1 and a biogenic amine increases EG3RF’s efficacy. Experimental results show that cysteamine is less harmful to cells and produces a greater decrease in calcium release (-52.77%) for EG3RF-transfected cells than propylamine (-21.90%). To further explore EG3RF’s role in reducing bladder inflammation, next steps include identifying its intracellular location in transfected cells and confirming the effect of EG3RF activation on urothelial ATP release. <b>Funding:</b> R01DK117884 (Beckel), R01DK117383 (Xu)." @default.
• W4377029208 created "2023-05-19" @default.
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• W4377029208 date "2023-05-18" @default.
• W4377029208 modified "2023-09-29" @default.
• W4377029208 title "Urothelial Knock-in of the Engineered Chloride Channel EG3RF as a Potential Treatment for Cystitis-Induced Bladder Inflammation" @default.
• W4377029208 doi "https://doi.org/10.1124/jpet.122.150210" @default.
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