Majima Tsuyoshi, Funahashi Yasuhito, Takai Shun, Goins William F, Gotoh Momokazu, Tyagi Pradeep, Glorioso Joseph C, Yoshimura Naoki
1 Department of Urology, University of Pittsburgh School of Medicine , Pittsburgh, Pennsylvania.
2 Department of Urology, Nagoya University Graduate School of Medicine , Nagoya, Japan .
Hum Gene Ther. 2015 Nov;26(11):734-42. doi: 10.1089/hum.2015.026. Epub 2015 Aug 31.
Increased afferent excitability has been proposed as an important pathophysiology of interstitial cystitis/bladder pain syndrome (IC/BPS) and overactive bladder (OAB). In this study, we investigated whether herpes simplex virus (HSV) vectors encoding poreless TRPV1, in which the segment in C terminus of TRPV1 receptor is deleted, suppress bladder overactivity and pain behavior using a rat model of chemical cystitis. Replication-defective HSV vectors encoding poreless TRPV1 were injected into the bladder wall of adult female Sprague-Dawley rats. Additionally, recombinant HSV virus (vHG) vectors were injected as control. Cystometry (CMG) under urethane anesthesia was performed 1 week after viral injection to evaluate bladder overactivity induced by resiniferatoxin (RTx, a TRPV1 agonist). RTx-induced nociceptive behavior such as licking (lower abdominal licking) and freezing (motionless head-turning) was observed 2 weeks after viral injection. GFP expression in L4/L6/S1 dorsal root ganglia and the bladder as well as c-Fos-positive cells in the L6 spinal cord dorsal horn were also evaluated 2 weeks after viral injection. In CMG, the poreless TRPV1 vector-treated group showed a significantly smaller reduction in intercontraction intervals and voided volume after RTx infusion than the vHG-treated control group. The number of the RTx-induced freezing events was significantly decreased in the poreless TRPV1 group than in the vHG group, whereas there was no significant difference of the number of RTx-induced licking events between groups. The number of c-Fos-positive cells in the DCM and SPN regions of the L6 spinal dorsal horn was significantly smaller in the poreless TRPV1 group than in the vHG group. Our results indicated that HSV vector-mediated gene delivery of poreless TRPV1 had a therapeutic effect on TRPV1-mediated bladder overactivity and pain behavior. Thus, the HSV vector-mediated gene therapy targeting TRPV1 receptors could be a novel modality for the treatment of OAB and/or hypersensitive bladder disorders such as IC/BPS.
传入神经兴奋性增加被认为是间质性膀胱炎/膀胱疼痛综合征(IC/BPS)和膀胱过度活动症(OAB)的重要病理生理学特征。在本研究中,我们使用化学性膀胱炎大鼠模型,研究编码无孔TRPV1(TRPV1受体C末端片段缺失)的单纯疱疹病毒(HSV)载体是否能抑制膀胱过度活动和疼痛行为。将编码无孔TRPV1的复制缺陷型HSV载体注射到成年雌性Sprague-Dawley大鼠的膀胱壁中。此外,注射重组HSV病毒(vHG)载体作为对照。病毒注射1周后,在乌拉坦麻醉下进行膀胱测压(CMG),以评估树脂毒素(RTx,一种TRPV1激动剂)诱导的膀胱过度活动。病毒注射2周后,观察RTx诱导的伤害性行为,如舔舐(下腹部舔舐)和僵住(静止转头)。病毒注射2周后,还评估了L4/L6/S1背根神经节和膀胱中的绿色荧光蛋白(GFP)表达以及L6脊髓背角中的c-Fos阳性细胞。在CMG中,无孔TRPV1载体治疗组在RTx注入后收缩间期和排尿量的减少明显小于vHG治疗的对照组。无孔TRPV1组中RTx诱导的僵住事件数量明显少于vHG组,而两组之间RTx诱导的舔舐事件数量没有显著差异。无孔TRPV1组L6脊髓背角DCM和SPN区域的c-Fos阳性细胞数量明显少于vHG组。我们的结果表明,HSV载体介导的无孔TRPV1基因递送对TRPV1介导的膀胱过度活动和疼痛行为具有治疗作用。因此,靶向TRPV1受体的HSV载体介导的基因治疗可能是治疗OAB和/或IC/BPS等膀胱过敏疾病的一种新方法。