Novel mechanisms regulating protein interaction and pain

项目摘要 多达20%的人口将遭受持续6个月以上的慢性疼痛。慢性疼痛 及其潜在的病理生理学，可导致抑郁和其他衰弱的神经效应 虽然有治疗急性疼痛的有效方法，但慢性疼痛对大多数当前的治疗方法都具有抵抗力。 需要开发针对这些疼痛状态下的分子事件的新疗法。 神经病理性和持续性手术后疼痛的发生，至少部分是由于长期的 脊髓背角兴奋性突触传递增强痛觉信号的功能 (痛觉过敏)和无伤大雅的刺激引起疼痛(痛觉过敏)。这些突触事件具有许多共同的特征 已在较高中枢神经系统区域研究的神经元可塑性。其中许多更改都依赖于NMDAR 导致突触强度增加。在这些变化背后出现的一个机制是 突触功能是通过与EphB的直接分子相互作用而增强NMDAR功能 受体酪氨酸激酶。在我们已发表的工作的基础上，我们将测试EphB-NMDAR的假设 相互作用通过以下方式将NMDAR定向到突触，从而导致慢性疼痛状态的发展 在体外和小鼠体内表达野生型或突变型EphB2受体。为了检验这一假设，我们将确定 介导EphB-NDMAR相互作用的机制，表征分子和其他工具来破坏这一相互作用 并确定阻止EphB-NMDAR相互作用是否会缓解慢性疼痛。至 针对这些问题，我们将实现三个具体目标：1.确定NMDAR上的域 负责EphB-NMDAR的交互。2.验证VLK引导磷酸化的假设 EphB2上的Y504。3.确定VLK在疼痛可塑性中的功能意义。总的来说，这些 AIMS将创造一种新的知识，使人们对EphB-NMDAR的作用有更深入的了解 疼痛中的相互作用并使理解慢性疼痛背后的基本机制取得进展 各州。

Project Abstract As much as 20% of the population will suffer from chronic pain lasting for more than 6 months. Chronic pain and its underlying pathophysiology, can result in depression and other debilitating neurological effects and although there are effective treatments for acute pain chronic pain is resistant to most current treatments requiring the development of novel therapeutics that target molecular events underlying these pain states. Neuropathic and persistent post-surgical pain occurs, at least in part, due to long lasting changes in the function of excitatory synaptic transmission in the spinal dorsal horn resulting in enhanced pain signaling (hyperalgesia) and innocuous stimuli evoking pain (allodynia). These synaptic events share many features of neuronal plasticity that has been studied in higher CNS areas. Many of these changes are NMDAR dependent resulting in increased synaptic strength. One mechanism that has emerged underlying these changes in synaptic function is the potentiation of NMDAR function by a direct molecular interaction with the EphB receptor tyrosine kinase. Building on our published work, we will test the hypothesis that an EphB-NMDAR interaction is responsible for the development of a chronic pain state by directing NMDARs to synapses by expressing wild type or mutant EphB2 receptors in vitro and in mice. To test this hypothesis, we will determine the mechanism mediating the EphB-NDMAR interaction, characterize molecules and other tools to disrupt this interaction, and determine whether preventing the EphB-NMDAR interaction will alleviate chronic pain. To address these questions we will undertake three specific aims: 1. Determine the domain on the NMDAR responsible for the EphB-NMDAR interaction. 2. Test the hypothesis that VLK directs phosphorylation of Y504 on EphB2. 3. Determine the functional significance of VLK in pain plasticity. Collectively these aims will create a new knowledge that will provide a deeper understanding of the role of EphB-NMDAR interaction in pain and enable progress toward understanding the basic mechanisms behind chronic pain states.