AB317. SPR-44 Pharmacological activation of individual KCNQ channel subtypes in detrusor smooth muscle represents a promising novel approach for overactive bladder treatment

Objective: Our recent studies have demonstrated voltagegated KCNQ channels (KCNQ1-KCNQ5) as key regulators of detrusor smooth muscle (DSM) function. Despite emerging developments, the physiological role of individual KCNQ channel subtypes remains less clear. Here, we utilized the novel compound ML-213, a potent activator of KCNQ2, KCNQ4, and KCNQ5 channels, to elucidate their physiological roles in guinea pig DSM function. Methods: Using isometric DSM tension recordings, Ca imaging, and amphotericin-B perforated patch-clamp electrophysiology, we elucidated the role of ML-213sensitive KCNQ channels in regulating DSM excitability and contractility. Results: ML-213 concentration-dependently (100 nM– 30 μM) inhibited spontaneous phasic, pharmacologicallyinduced, and nerve-evoked contractions in DSM isolated strips. ML-213 (10 μM) decreased the global intracellular Ca concentrations in DSM isolated strips, effects blocked by the L-type voltage-gated Ca (CaV) channel inhibitor nifedipine (1 μM) and the KCNQ1-KCNQ5 channel inhibitor XE991 (10 μM). These data suggest that ML213 decreases the global intracellular Ca concentration by inhibiting L-type CaV channels through an indirect mechanism downstream from KCNQ channel activation. In addition, ML-213 hyperpolarized the cell membrane potential and inhibited spontaneous action potentials in DSM cells, effects reversible by washout. We next aimed to examine the effects of ML-213 on whole cell KCNQ currents. To isolate KCNQ currents, the bath solution contained the large conductance voltageand Ca-activated K channel inhibitor paxilline (1 μM) and gadolinium chloride (GdCl3, 50 μM), which blocks L-type CaV channels and non-selective cation channels. Under these experimental conditions, ML-213 (10 μM) enhanced whole cell KCNQ currents. These findings suggest that the modulation of K transport through ML-213-sensitive KCNQ channels underlies ML-213-induced cell membrane hyperpolarization to decrease the global intracellular Ca concentration and DSM contractility. Conclusions: These data using the novel compound ML-213 , sugges t tha t KCNQ2, KCNQ4, and KCNQ5-containing channels are essential regulators of the excitability, intracellular Ca concentration, and contractility of DSM by virtue of their control of the membrane potential. Moreover, these new findings provide a foundational basis for future investigations on KCNQ channel functional roles in human DSM excitability and contractility to confirm their potential as novel therapeutic targets for overactive bladder. Funding Source(s): NIH grant R01-DK106964 to GV Petkov and F31-DK104528 to A Provence.