Joseph S. Cefalu

Purinoceptors as therapeutic targets for lower urinary tract dysfunction

Lower urinary tract symptoms (LUTS) are present in many common urological syndromes. However, their current suboptimal management by muscarinic and α1‐adrenoceptor antagonists leaves a significant opportunity for the discovery and development of superior medicines. As potential targets for such therapeutics, purinoceptors have emerged over the last two decades from investigations that have established a prominent role for ATP in the regulation of urinary bladder function under normal and pathophysiological conditions. In particular, evidence suggests that ATP signaling via P2X1 receptors participates in the efferent control of detrusor smooth muscle excitability, and that this function may be heightened in disease and aging. ATP also appears to be involved in bladder sensation, via activation of P2X3 and P2X2/3 receptors on sensory afferent neurons, both within the bladder itself and possibly at central synapses. Such findings are based on results from classical pharmacological and localization studies in non‐human and human tissues, knockout mice, and studies using recently identified pharmacological antagonists – some of which possess attributes that offer the potential for optimization into candidate drug molecules. Based on recent advances in this field, it is clearly possible that the development of selective antagonists for these receptors will occur that could lead to therapies offering better relief of sensory and motor symptoms for patients, while minimizing the systemic side effects that limit current medicines.

Endogenous Purinergic Control of Bladder Activity via Presynaptic P2X3 and P2X2/3 Receptors in the Spinal Cord

P2X3 and P2X2/3 receptors are localized on sensory afferents both peripherally and centrally and have been implicated in various sensory functions. However, the physiological role of these receptors expressed presynaptically in the spinal cord in regulating sensory transmission remains to be elucidated. Here, a novel selective P2X3 and P2X2/3 antagonist, AF-792 [5-(5-ethynyl-2-isopropyl-4-methoxy-phenoxy)-pyrimidine-2,4-diamine, previously known as RO-5], in addition to less selective purinoceptor ligands, was applied intrathecally in vivo. Cystometry recordings were made to assess changes in the micturition reflex contractions after drug treatments. We found that AF-792 inhibited micturition reflex activity significantly (300 nmol; from baseline contraction intervals of 1.18 ± 0.07 to 9.33 ± 2.50 min). Furthermore, inhibition of P2X3 and P2X2/3 receptors in the spinal cord significantly attenuated spinal activation of extracellular-signal regulated kinases induced by acute peripheral stimulation of the bladder with 1% acetic acid by 46.4 ± 12.0% on average. Hence, the data suggest that afferent signals originating from the bladder are regulated by spinal P2X3 and P2X2/3 receptors and establish directly an endogenous central presynaptic purinergic mechanism to regulate visceral sensory transmission. Identification of this spinal purinergic control in visceral activities may help the development of P2X3 and P2X2/3 antagonist to treat urological dysfunction, such as overactive bladder, and possibly other debilitating sensory disorders, including chronic pain states.

Selective Pharmacological Blockade of the TRPV1 Receptor Suppresses Sensory Reflexes of the Rodent Bladder

PURPOSE We investigated the pharmacological effect of TRPV1 antagonists in anesthetized rodent models of bladder function. MATERIALS AND METHODS The TRPV1 antagonists JNJ17203212 and JYL1421 were evaluated in the anesthetized rat volume induced micturition reflex model. JNJ17203212 was further evaluated in this model in capsaicin (Sigma) desensitized rats, and in rat capsaicin and mouse citric acid models of irritant induced detrusor overactivity. RESULTS Systemic JNJ17203212 and JYL1421 administration in the anesthetized rat volume induced micturition reflex model resulted in an increased micturition threshold volume. JNJ17203212 also decreased bladder contraction amplitude but JYL1421 had no effect. Capsaicin desensitization significantly increased baseline micturition threshold volume and decreased bladder contraction amplitude in the volume induced micturition reflex model compared to those in sham treated controls and JNJ17203212 produced no further effect after capsaicin desensitization. JNJ17203212 was also effective in 2 models of irritant induced detrusor overactivity, preventing the decrease in micturition threshold volume and the increase in bladder contraction amplitude observed with intravesical instillation of 10 microM capsaicin, and the decreased voiding interval induced by intravesical citric acid. CONCLUSIONS The TRPV1 antagonists JNJ17203212 and JYL1421 increased the threshold for activation of the micturition reflex in the anesthetized rat volume induced micturition reflex model. This effect appeared to be mediated by capsaicin sensitive afferents. JNJ17203212 also inhibited detrusor overactivity induced by intravesical capsaicin and intravesical citric acid. These data extend our understanding of the role of TRPV1 in sensory modulation of the micturition reflex under nonirritant and inflammatory conditions.

Near-Field Ultrasound Imaging During Radiofrequency Catheter Ablation: Tissue Thickness and Epicardial Wall Visualization and Assessment of Radiofrequency Ablation Lesion Formation and Depth

Background Safe and successful radiofrequency catheter ablation depends on creation of transmural lesions without collateral injury to contiguous structures. Near-field ultrasound (NFUS) imaging through transducers in the tip of an ablation catheter may provide important information about catheter contact, wall thickness, and ablation lesion formation. Methods and Results NFUS imaging was performed using a specially designed open-irrigated radiofrequency ablation catheter incorporating 4 ultrasound transducers. Tissue/phantom thickness was measured in vitro with varying contact angles. In vivo testing was performed in 19 dogs with NFUS catheters positioned in 4 chambers. Wall thickness measurements were made at 222 sites (excluding the left ventricle) and compared with measurements from intracardiac echocardiography. Imaging was used to identify the epicardium with saline infusion into the pericardial space at 39 sites. In vitro, the measured exceeded actual tissue/phantom thickness by 13% to 20%. In vivo, NFUS reliably visualized electrode-tissue contact, but sensitivity of epicardial imaging was 92%. The chamber wall thickness measured by NFUS correlated well with intracardiac echocardiography (r=0.86; P<0.0001). Sensitivity of lesion identification by NFUS was 94% for atrial and 95% for ventricular ablations. NFUS was the best parameter to predict lesion depth in right and left ventricle (r=0.47; P<0.0001; multiple regression P=0.0025). Lesion transmurality was correctly identified in 87% of atrial lesions. Conclusions NFUS catheter imaging reliably assesses electrode-tissue contact and wall thickness. Its use during radiofrequency catheter ablation may allow the operator to assess the depth of ablation required for transmural lesion formation to optimize power delivery.