NOVEL MODULATORS OF THE VANILLOID RECEPTOR

香草酸受体（TRPV 1;以前称为VR 1）在原发性痛觉过敏中起重要作用， 慢性疼痛的组成部分。TRPV 1在病理生理状态如疼痛， 糖尿病性神经病变和膀胱病。TRPV 1的激活通过许多会聚信号传导发生， 途径和新出现的证据表明，TRPV 1的功能也受到许多 机制等TRPV 1功能的新型细胞抑制剂或负调节剂的发现及其应用 这种作用方式应该为TRPV 1在外周疼痛信号传导中的作用提供额外的见解， 延伸，建议新的方法，其应用程序控制原发性痛觉过敏。我们提出 为了检验以下假设：（i）新的细胞产物在抑制或阴性表达中起重要作用， TRPV 1功能的调节和（ii）这些分子的基于载体的表达可以控制TRPV 1 体内信号传导为了验证这些假设，我们设计了一种基于HSV载体的遗传学组合， 识别这些潜在调控分子的策略和表征其作用的生化方法 在TRPV 1功能的阴性对照中。具体而言，我们将尝试确定产品，（a）抑制 辣椒素（CAP）和树脂毒素（RTX）对TRPV 1的激活和（B）通过以下方式干扰TRPV 1的增强作用： 由佛波醇12-肉豆蔻酸酯13-乙酸酯（PMA）激活的蛋白激酶C β（PKCe）。TRPV 1抑制剂 基因，无论是已经工程或自然发生，将进行详细研究，以确定 它们能够阻止TRPV 1介导的钙内流的分子机制。四个具体目标 我们打算（i）建立模型系统，其中HSV TRPV 1表达载体可用于检查 TRPV 1功能或钙超载的潜在抑制剂（ii）创建表达cDNA的载体库 来源于大鼠背根神经节，（iii）表征所选神经元基因 产品抑制或负性调节TRPV 1功能，和（iv）评价产品的镇痛作用。 使用大鼠疼痛模型体内工程化的和新的TRPV 1抑制剂，其中TRPV 1拮抗作用已被 减少疼痛信号。模型抑制基因和通过HSV感染获得的新基因都可以用于研究HSV感染的抑制基因。 筛选方法将提供给项目1和2，以评估其潜在的镇痛作用， 糖尿病相关的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.