Peripheral Neuronal and Non-neuronal Mechanisms of Fabry Disease Pain

项目总结 Fabry病(FD)是最常见的溶酶体储存疾病之一，可引起严重的疼痛。 患者从很小的时候就开始。虽然临床文献显示，患有FD的患者有机械性- 诱发性疼痛、持续性疼痛和周围神经损伤，目前尚不清楚FD疼痛是如何在 周围神经系统(PNS)。我的提案将研究三叉神经节细胞机制如何介导FD疼痛 使用FD大鼠模型，该模型概括了患者的疼痛表型。异常离子通道 在许多疾病中，神经元上的活动与外周介导的疼痛有关。然而，最近的证据表明 研究表明，神经元中的疼痛相关离子通道也在非神经元细胞中表达。递减 雪旺细胞(SC)离子通道活动可改善其他疼痛的机械性疼痛行为 条件。已有假说认为，从SC释放的变应原正在影响机械诱发的 疼痛。我们已经证明，FD中机械诱发的疼痛依赖于离子通道瞬变的活动 三叉神经节内的受体电位Ankyrin 1(TRPA1)。在我的提案中，我将确定TRPA1是否也参与调解 功能性消化不良大鼠持续疼痛通过三七总皂甙活性。我还将确定FD SC释放的变应原是否激活 或者使神经元对机械力敏感，如果SC TRPA1在FD中驱动机械诱发的疼痛。我 假设TRPA1在雪旺细胞和神经元中的活性增加对 维持FD大鼠的慢性疼痛表型。在目标1中，我将确定FD大鼠是否有行为 持续疼痛和依赖于TRPA1的异常周围神经活动。我会用一块正在进行的电池 疼痛行为技术，并确定TRPA1抑制是否减轻这种疼痛(目标1A)。到时候我会决定 外周TRPA1是否参与FD大鼠的自发性轴突活动 光纤记录(目标1B)。在目标2中，我将研究SCs在依赖TRPA1的机械过程中所起的作用 FD的超敏反应。首先，我将确定感觉神经元是否被变应原激活或敏化 从FD SCs释放，以及这种激活是否需要神经元TRPA1(目标2A)。接下来，我将确定是否 来自FD SCs的抗原原通过膜片钳记录使神经元对机械刺激敏感，如果这 取决于神经元的TRPA1活性(目标2B)。最后，我将确定SC TRPA1的病毒抑制是否可以 通过一系列疼痛行为分析减轻FD的机械过敏(目标2C)。我的提议将 为雪旺细胞如何介导慢性疼痛提供了基础，并将促进我们对 法布里病痛。

PROJECT SUMMARY Fabry Disease (FD) is one of the most common lysosomal storage diseases and causes devastating pain in patients starting at a young age. While clinical literature has shown that patients with FD have mechanically- evoked pain, ongoing pain, and peripheral nerve damage, it is unclear how FD pain is mediated in the peripheral nervous system (PNS). My proposal will investigate how PNS cellular mechanisms mediate FD pain using the FD rat model, which recapitulates the pain phenotypes seen in patients. Abnormal ion channel activity on neurons is linked to peripherally mediated pain in many diseases. However, recent evidence has shown that pain-associated ion channels in neurons are also expressed in non-neuronal cells. Decreasing Schwann cell (SC) ion channel activity can ameliorate mechanically-evoked pain behaviorally in other pain conditions. It has been hypothesized that algogens released from SCs are influencing mechanically-evoked pain. We have shown that mechanically-evoked pain in FD depends on the activity of the ion channel Transient Receptor Potential Ankyrin 1 (TRPA1) in the PNS. In my proposal, I will determine if TRPA1 also mediates ongoing pain through PNS activity of FD rats. I will also determine if algogens released from FD SCs activate or sensitize neurons to mechanical force, and if SC TRPA1 drives mechanically-evoked pain in FD. I hypothesize that increased activity of TRPA1 in both Schwann cells and neurons is critical for maintaining chronic pain phenotypes in the FD rat. In Aim 1, I will determine if the FD rat has behavioral ongoing pain and aberrant peripheral nerve activity that is dependent on TRPA1. I will use a battery of ongoing pain behavior techniques and determine if TRPA1 inhibition alleviates this pain (Aim 1A). I will then determine whether peripheral TRPA1 is required for spontaneous axonal activity in the FD rat by using ex vivo teased fiber recordings (Aim 1B). In Aim 2, I will investigate the role that SCs play in TRPA1-dependent mechanical hypersensitivity in FD. First, I will determine if sensory neurons are activated or sensitized by algogens released from FD SCs, and if neuronal TRPA1 is required for this activation (Aim 2A). Next, I will determine if algogens from FD SCs sensitize neurons to mechanical stimulation using patch-clamp recordings, and if this depends on neuronal TRPA1 activity (Aim 2B). Finally, I will determine if viral inhibition of SC TRPA1 can alleviate FD mechanical hypersensitivity through a battery of pain behavior assays (Aim 2C). My proposal will provide a basis for how Schwann cells mediate chronic pain and will advance our understanding of Fabry Disease pain.