NOVEL MODULATORS OF THE VANILLOID RECEPTOR

香草酸受体(TRPV1；以前称为VR1)在原发性痛觉过敏中起着重要作用。 慢性疼痛的组成部分。TRPV1在疼痛等病理生理状态下功能上调 糖尿病神经病变和膀胱病。TRPV1的激活通过多个汇聚信号发生 通路和新的证据表明，TRPV1的功能也受到许多负调控 机械装置。TRPV1功能的新型细胞抑制或负调制子的发现及其意义 行动模式应该为TRPV1在外周疼痛信号中的作用提供更多的见解，并通过 推广，提出了新的方法来应用于控制原发性痛觉过敏。我们建议 检验假设：(I)新的细胞产物在抑制或负面作用中起重要作用 调节TRPV1功能和(Ii)这些分子的基于载体的表达可以控制TRPV1 体内的信号。为了验证这些假设，我们设计了一种基于HSV载体的基因组合 确定这些潜在调控分子的策略和确定其作用的生化方法 在对TRPV1功能的负性调控中。具体地说，我们将尝试识别(A)抑制的产品 辣椒素(CAP)和树脂毒素(RTX)激活TRPV1和(B)通过以下途径干扰TRPV1增强 蛋白激酶C epsilon(PKCE)由佛波醇12-肉豆蔻酸酯13-乙酸酯(PMA)激活。TRPV1抑制 我们将对经过改造或自然产生的基因进行更详细的研究，以确定 它们能够阻止TRPV1介导的钙内流的分子机制。在四个具体目标中 我们打算(I)建立模型系统，在其中HSV TRPV1表达载体可用于检测 TRPV1功能或钙超载的潜在抑制剂(II)创建表达cDNA的载体文库 来源于大鼠背根神经节，(Iii)表征了所选神经基因的机制 产品抑制或负向调节TRPV1的功能和(Iv)评估 利用TRPV1拮抗的大鼠疼痛模型进行体内工程和新型TRPV1抑制剂的研究 显示可以减少疼痛信号。单纯疱疹病毒模型抑制基因和新基因 将向项目1和项目2提供筛选方法，以评估其潜在的止痛效果。 糖尿病相关神经性疼痛和膀胱疼痛模型。

The vanilloid receptor (TRPV1; formerly known as VR1) plays an important role in primary hyperalgesia, a component of chronic pain. TRPV1 is functionally up-regulated in patho-physiologic states such as painful diabetic neuropathy and cystopathy. Activation of TRPV1 occurs through a number of convergent signaling pathways and emerging evidence suggests that TRPV1 function is also negatively regulated by numerous mechanisms. The discovery of novel cellular inhibitors or negative modulators of TRPV1 function and their mode of action should provide additional insights into the role of TRPV1 in peripheral pain signaling and by extension, suggest novel approaches for their application to the control of primary hyperalgesia. We propose to test the hypotheses that (i) novel cellular products play an important role in the inhibition or negative modulation of TRPV1 function and (ii) that vector-based expression of these molecules can control TRPV1 signaling in vivo. To test these hypotheses, we have devised a combination of HSV vector-based genetic strategies to identify these potential regulatory molecules and biochemical methods to characterize their role in the negative control of TRPV1 function. Specifically, we will attempt to identify products that (a) inhibit TRPV1 activation by capsaicin (CAP) and resiniferatoxin (RTX) and (b) interfere with TRPV1 potentiation by protein kinase C epsilon (PKCe) activated by Phorbol 12-myristate 13-acetate (PMA). TRPV1 inhibitory genes that either have been engineered or naturally occur, will be studied in some detail to determine the molecular mechanism underlying their ability to impede TRPV1-mediated calcium influx. In four specific aims we intend to (i) create model systems in which HSV TRPV1 expression vectors can be used to examine potential inhibitors of TRPV1 function or calcium overload (ii) create a library of vectors expressing cDNAs derived from rat dorsal root ganglia, (iii) characterize mechanisms by which the selected neuronal gene products inhibit or negatively modulate TRPV1 function and (iv) evaluate the analgesic effects of the engineered and novel TRPV1 inhibitors in vivo using rat models of pain where TRPV1 antagonism has been shown to reduce pain signaling. Both the model inhibitor genes and novel genes obtained by the HSV screening methods will be provided to Projects 1 and 2 for evaluation of their potential analgesic effects in diabetes-related neuropathic pain and models of bladder pain.