Integrative And Molecular Studies Of Pain And Pain Contr

疼痛和疼痛控制的综合和分子研究

• 批准号: 6966492
• 负责人: Michael J. Iadarola
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF DENTAL & CRANIOFACIAL RESEARCH
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6966492
• 关键词: analgesia; central nervous system; clinical trials; combinatorial chemistry; gene therapy; genetic transcription; human subject; molecular cloning; neural plasticity; neural transmission; neuropeptides; patient oriented research; proteomics; tissue /cell culture

Our research program addresses basic molecular and physiological processes of nociceptive transmission in the central nervous system and new, effective ways to treat intractable pain. The molecular research is performed using animal and in vitro cell-based models. We concentrate on primary afferent pain-sensing neurons that innervate the skin and deep tissues and their connections in the dorsal spinal cord, which is the first site of synaptic information processing for pain. Our research has identified it as a locus of neuronal plasticity and altered gene expression in persistent pain states. The regulation of transduction of physical pain stimuli is also under investigation using cloned thermal and chemo-responsive ion channels ectopically expressed in heterologous cell systems and naturally expressed in primary cultures of dorsal root ganglion. Our goals are (1) to understand the molecular and cell biological mechanisms of acute and chronic pain at these two basic levels of the nervous system and (2) to use this knowledge to devise new treatments for pain. We address the latter goal in a translational research and human clinical trials program designed to evaluate new analgesic treatment for severe pain. The treatment we are developing is based on our studies of the molecular mechanisms of pain transduction through the vanilloid receptor 1 (TRPV1). This molecule is a heat-sensing calcium/sodium ion channel that converts painful heat into nerve action potentials by opening the channel and depolarizing pain-sensing nerve terminals. Channel opening is also stimulated by capsaicin which is a vanilloid chemical and the active ingredient in hot pepper. We use a very potent vanilloid analog to prop open the channel causing death of a specific class of pain-sensing neurons, yet allowing mechanical and high temperature heat pain sensations and other somatosensory and proprioceptive sensations to remain intact. We have established an inter-institute working group with NIDA's Division of Pharmaco-Therapeutics and Medical Consequences of Drug Abuse to bring the treatment to human clinical trial. The working group consists of experts on medical, neurobiological, toxicological, chemical and formulation issues as well as anesthesiologists, pharmacologists and pathologists from our group. Over the course of this year, we also established mechanisms for obtaining the natural product from which the active drug is extracted and procedures for isolation, purification and formulation of the drug product such that it will be compliant with Food and Drug Administration (FDA) regulations. We are presently finalizing the toxicology study, the Investigational New Drug Application with the FDA and the Human Clinical Protocol with the NCI's IRB. The treatment we have devised may be a very effective approach to control many types of chronic pain especially those associated with cancer, arthritis, tempromandibular joint disorders, trigeminal neuralgia and chronic neuropathic pain problems. Underlying the translational studies are the questions of molecular regulation in chronic pain and mechanisms of pain transduction in peripheral nerve ending. These questions are addressed using subtraction cloning, differential hybridization and gene arrays, and neurophysiological measurements such as calcium imaging in live cells. The physiological stdies have focused on he multiple intracellular pools of calcium that can be activated by vanilloid agonists and the interaction of these pools with the plasma membrane localized TRPV1 and TRPV1 located on the endoplasmic reticulum. Activation of TRPV1 in both locations is a factor that underlies the efficacy of TRPV1 agonists at inducing calcium cytotoxicity in the above translational studies. The molecular studies reveal a more dynamic modulation of gene expression in dorsal root ganglion than previously hypothesized, for example, in a matter of hoours we observe up-regulation of the receptor for Neuropeptide FF, which is known to be involved in opioid modulation of pain. In addition to pain, these studies fundamentally explore the molecular basis of synaptic plasticity. New roles for the calcium and arachidonic acid binding proteins S100A8 and S100A9 in spinal cord and dorsal root ganglion have also been discovered. We hypothesize modularity in the neuronal response new levels of synaptic or pharmacological input (e.g. learning, neurological disorders, drug abuse). The "generic" alterations are combined with modulation of tissue-specific genes to meet the demands generated by the new level of stimulation. This will lead to a deeper understanding of molecular mechanisms that trigger and sustain chronic pain.

我们的研究项目涉及中枢神经系统伤害性传递的基本分子和生理过程，以及治疗顽固性疼痛的新的有效方法。分子研究使用动物和体外细胞模型进行。我们专注于初级传入疼痛感觉神经元，支配皮肤和深部组织及其在背侧脊髓中的连接，这是疼痛突触信息处理的第一个站点。我们的研究已经确定它是神经元可塑性的一个位点，并在持续性疼痛状态下改变基因表达。物理疼痛刺激的转导的调节也正在调查中使用克隆的热和化学反应离子通道异位表达在异源细胞系统和自然表达在背根神经节的原代培养物。我们的目标是（1）了解神经系统这两个基本水平的急性和慢性疼痛的分子和细胞生物学机制，（2）利用这些知识设计新的疼痛治疗方法。我们在翻译研究和人体临床试验项目中解决了后一个目标，该项目旨在评估用于严重疼痛的新镇痛治疗。我们正在开发的治疗方法是基于我们对通过香草素受体1（TRPV 1）进行疼痛转导的分子机制的研究。这种分子是一种热感应钙/钠离子通道，通过打开通道和去极化疼痛感应神经末梢，将疼痛的热量转化为神经动作电位。辣椒素也能刺激通道开放，辣椒素是一种香草素化学物质，也是辣椒中的活性成分。我们使用一种非常有效的香草素类似物来支撑打开通道，导致特定类别的疼痛感受神经元死亡，但允许机械和高温热疼痛感觉以及其他躯体感觉和本体感觉保持完整。我们已经与NIDA药物治疗和药物滥用的医学后果部门建立了一个机构间工作组，以将治疗方法引入人体临床试验。该工作组由医学、神经生物学、毒理学、化学和制剂问题专家以及我们小组的麻醉师、药理学家和病理学家组成。在这一年中，我们还建立了获得从中提取活性药物的天然产品的机制，以及药物产品的分离、纯化和配制程序，以符合食品药品监督管理局（FDA）的规定。我们目前正在完成毒理学研究，FDA的研究性新药申请和NCI IRB的人类临床方案。我们设计的治疗方法可能是一种非常有效的方法来控制许多类型的慢性疼痛，特别是那些与癌症，关节炎，颞下颌关节疾病，三叉神经痛和慢性神经性疼痛问题有关的疼痛。 转译研究的基础是慢性疼痛的分子调控和外周神经末梢疼痛传导机制的问题。这些问题是解决使用减法克隆，差异杂交和基因阵列，和神经生理学测量，如活细胞中的钙成像。生理学研究主要集中在可被香草素激动剂激活的多个细胞内钙池以及这些钙池与质膜定位的TRPV 1和位于内质网上的TRPV 1的相互作用。在上述转化研究中，TRPV 1在两个位置的激活是TRPV 1激动剂诱导钙细胞毒性的功效的基础因素。分子研究揭示了背根神经节中基因表达的动态调节比以前假设的更强，例如，在几个小时内，我们观察到神经肽FF受体的上调，已知其参与阿片类药物对疼痛的调节。除了疼痛，这些研究还从根本上探索了突触可塑性的分子基础。钙和花生四烯酸结合蛋白S100 A8和S100 A9在脊髓和背根神经节中的新作用也被发现。我们假设神经元反应的模块化新水平的突触或药理学输入（如学习，神经系统疾病，药物滥用）。“通用”改变与组织特异性基因的调节相结合，以满足新水平刺激产生的需求。这将导致对引发和维持慢性疼痛的分子机制的更深入理解。