Cystitis-induced bladder dysfunction and pain

项目摘要/摘要 膀胱痛综合征(BPS)/间质性膀胱炎(IC)是一种慢性盆腔疼痛疾病，至少有一个尿路 症状和疼痛起源于膀胱的感觉。压力会加剧 Bps/IC。尽管进行了密集的研究，但我们缺乏对心脏结构和功能变化的了解 排尿反射与BPS/IC有关，以及压力如何加剧症状，从而阻碍疗效 治疗。应对这些挑战部分需要：(1)涉及NGF/TrkA/MAPK的新假说 下游瞬时受体电位阳离子通道亚家族香草素成员4的信号转导 排尿反射感觉成分中的(TRPV4)/钙激活；(2)创新和多学科 方法；和(3)总结BPS/IC临床症状/体征的动物模型，包括症状 由心理压力引起的恶化(耀斑)。我们的实验室在互补用途上是独一无二的 几个相关的模型来加强我们的研究，包括(例如，环磷酰胺(CYP)诱导的膀胱炎， 慢性尿路上皮NGF高表达转基因小鼠(NGF-OE)，重复低剂量CYP(单独) 不足以产生显著症状)与重复变量应激(RVS)相结合来评估压力如何 会加重疾病。我们的总体假设是尿频和盆腔感觉的增加 伴发BPS/IC反映神经化学介质表达、功能和相互作用增强 感觉转导，TRPV4，在排尿反射的感觉成分中，有利于前兴奋性 州政府。在我们以前工作的基础上，NGF/TrkA/p75NTR信号和下游的适应不良角色(S) 在排尿反射的感觉成分中，TRPV4/钙离子的激活将被评估为有贡献的 BPS/IC的机制。目标1：确定阻断NGF/TrkA/p75NTR信号通路是否减少排尿 频率和盆腔疼痛：通过减少(1)尿上皮钙离子事件；(2)尿上皮ATP释放和(3)膀胱 传入活动。进一步中断NGF/p75NTR信号通过：(1)减少排尿频率 尿路上皮细胞凋亡；(2)促进尿路上皮细胞恢复；(3)维持跨上皮抵抗。 目的2：确定排尿途径中NGF信号的中断是否具有短期和长期的影响 BPS/IC样症状对TRPV4/钙离子功能的影响急性NGF介导的TRPV4/钙离子 与BPS/IC相关的反应包括尿路上皮细胞钙信号增强、尿路上皮细胞ATP分泌和膀胱 传入神经活动。适应不良的长期NGF信号通过增加TRPV4转录促进BPS/IC 和蛋白质的表达。使用三种具有BPS/IC样症状的模型和多学科、细胞-组织- 系统实验方法，我们将确定：(1)潜在的结构和功能变化 导致BPS/IC样症状；(2)心理应激对膀胱功能和疼痛的影响 (3)新的治疗方法。

Project Summary/Abstract Bladder Pain Syndrome (BPS)/Interstitial Cystitis (IC) is a chronic pelvic pain disorder with at least one urinary symptom and the perception that the pain originates from the bladder. Stress exacerbates symptoms of BPS/IC. Despite intense research, we lack understanding of how structural and functional changes in the micturition reflex are linked to BPS/IC and how stress exacerbates symptoms, thus impeding effective therapies. Addressing these challenges requires, in part: (1) a novel hypothesis involving NGF/TrkA/MAPK signaling for downstream transient receptor potential cation channel subfamily vanilloid member 4 (TRPV4)/Ca2+ activation in the sensory components of the micturition reflex; (2) innovative and multidisciplinary approaches; and (3) animal models that recapitulate the clinical signs/symptoms of BPS/IC including symptom exacerbation (flares) precipitated by psychological stress. Our laboratory is unique in the complementary use of several relevant models to reinforce our studies including (e.g., cyclophosphamide (CYP)-induced cystitis, transgenic mice with chronic, urothelial overexpression of NGF (NGF-OE), repeated, low dose CYP (alone insufficient to produce significant symptoms) coupled with repeated variate stress (RVS) to assess how stress can exacerbate disease. Our overall hypothesis is that increases in urinary frequency and pelvic sensation that accompany BPS/IC reflect increased expression, function and interactions of neurochemical mediators and the sensory transducer, TRPV4, in the sensory components of the micturition reflex that favor a pro-excitatory state. Building from our previous work, the maladaptive role(s) of NGF/TrkA/p75NTR signaling and downstream activation of TRPV4/Ca2+ in the sensory components of the micturition reflex will be assessed as contributory mechanisms to BPS/IC. Aim 1: Determine if interrupting NGF/TrkA/p75NTR signaling pathways reduces voiding frequency and pelvic pain by: reducing (1) urothelial Ca2+ events; (2) urothelial ATP release and (3) bladder afferent activity. Further interrupting NGF/p75NTR signaling reduces voiding frequency by: (1) reducing urothelial cell apoptosis; (2) promoting urothelial cell recovery and (3) maintaining transepithelial resistance. Aim 2: Determine if disruption of NGF signaling in the micturition pathway has short- and long-term consequences on TRPV4/Ca2+ function in BPS/IC-like symptoms. The acute NGF-mediated TRPV4/Ca2+ BPS/IC-related responses include heightened urothelial Ca2+ signaling, urothelial ATP secretion and bladder afferent nerve activity. Maladaptive, long-term NGF signaling promotes BPS/IC by increasing TRPV4 transcript and protein expression. Using three models with BPS/IC-like symptoms and a multidisciplinary, cell-tissue- systems experimental approach, we will determine: (1) underlying structural and functional changes contributory to BPS/IC-like symptoms; (2) the influence of psychological stress on bladder function and pain and (3) novel treatments.