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.

我们的研究计划介绍了中枢神经系统中伤害感受传播的基本分子和生理过程，以及治疗顽固性疼痛的新方法。分子研究是使用动物和基于体外细胞的模型进行的。我们集中于一级传入疼痛神经元，该神经元支配皮肤和深层组织及其在背脊髓中的连接，这是突触信息处理疼痛的第一个部位。我们的研究将其确定为神经元可塑性的一个轨迹，并改变了持续性疼痛状态的基因表达。还使用克隆的热和化学反应离子通道在异源细胞系统中表达并在背侧根神经节的一级培养中自然表达，从而调查了身体疼痛刺激的转导刺激。我们的目标是（1）了解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，以及（2）使用这些知识来设计新的疼痛治疗方法。我们解决了一项转化研究和人类临床试验计划中的后一种目标，旨在评估新的镇痛治疗方法。我们正在开发的处理是基于我们对通过香草素受体1（TRPV1）的疼痛转导的分子机制的研究。该分子是一种热感应钙/钠离子通道，通过打开通道并去极化疼痛感应神经末端将疼痛的热量转化为神经动作电位。辣椒素也刺激了通道的开口，辣椒素是香草胶化学物质和热胡椒中的活性成分。我们使用非常有效的香草型模拟来打开导致特定类型疼痛神经元死亡的通道，但允许机械和高温热疼痛感觉以及其他体感和本体感受的感觉保持完整。我们已经建立了一个由NIDA的药物治疗师和药物滥用的医疗后果的部门工作组，以将治疗方法带入人类临床试验。工作组由我们小组的医学，神经生物学，毒理学，化学和配方问题以及麻醉师，药理学家和病理学家的专家组成。在今年的整个过程中，我们还建立了获取活性药物的天然产物的机制，并进行了隔离，纯化和制定药物的方法，以使其符合食品和药物管理（FDA）法规。我们目前正在最终确定毒理学研究，即FDA的研究新药和NCI IRB的人类临床方案。我们设计的治疗方法可能是控制多种类型的慢性疼痛的一种非常有效的方法，尤其是与癌症，关节炎，临时颌骨关节疾病，三叉神经痛和慢性神经性疼痛问题有关的方法。 翻译研究的基础是分子调节在慢性疼痛和周围神经结尾的疼痛转导机制中的问题。这些问题是使用减法克隆，差异杂交和基因阵列以及神经生理测量值（例如活细胞中的钙成像）来解决的。生理性迹象集中在HE多个细胞内钙池上，这些钙可以被香草酸激动剂激活，以及这些池与位于内质网上上的质膜局部TRPV1和TRPV1的相互作用。在这两个位置，TRPV1的激活是在上述转化研究中诱导钙细胞毒性诱导钙的钙的疗效的基础的一个因素。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，例如，在houour的问题中，我们观察到神经肽FF受体的上调，该受体已知与阿片类疼痛有关。除了疼痛外，这些研究从根本上探索了突触可塑性的分子基础。还发现了脊髓和背根神经节中钙和花生四烯酸结合蛋白S100A8和S100A9的新作用。我们假设神经元反应中的模块化新水平的突触或药理输入水平（例如学习，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，以满足新刺激水平产生的需求。这将导致对触发和维持慢性疼痛的分子机制有更深入的了解。