AAV-encoded analgesic peptide aptamers for chronic pain

 说明： 慢性疼痛是延误受伤后康复的关键限制因素。因为目前 在现有治疗方法不足的情况下，开发新的治疗方法是国家医疗保健的优先事项。多年的动物实验已经产生了大量的机制观察，在细胞和分子水平上解释了疼痛的开始和延续，但这种努力还没有出现新的治疗方法。两个关键的限制是典型的小分子药物的非特异性作用，以及当药物系统地给药时，无法限制由非靶标行为引起的危险副作用。我们设计了一种方案，在将重组腺相关病毒(AAV)载体注射到背根后，在感觉神经元中表达小的止痛肽(称为肽适配子)。 神经节(DRG)。大量数据表明，背根神经节水平的注射对动物和人类都是安全的。这种方法也只使用极小剂量的药物，并将效果限制在该节段的目标外周感觉神经元。这些肽适配子的设计目的是干扰关键分子靶标的组装、运输或功能，这些分子靶标调节外周感觉转导和背角(DH)突触功能，导致高度选择性的外周调节感觉神经元的性能。这最后一个特征是至关重要的，因为慢性疼痛的发生和持续显然涉及到外周受体和传递疼痛的突触的可塑性。本项目的总体目标是定义一种新的策略，即AAV递送的止痛肽适配子，用于选择性地调节初级感觉神经元中特定伤害性通路的细胞内蛋白质相互作用，并测试该方法作为治疗慢性疼痛的分子治疗工具的有效性。建议的研究将测试两个关键疼痛信号节点的概念验证，即N型钙通道(Cav2.2)和瞬时受体潜在香草素1通道(TRPV1)。这项拟议研究的成功完成将为开发一种全新的、功能强大的节段性慢性疼痛治疗模式奠定基础，从而将大量积压的机械发现转化为实际治疗。

 DESCRIPTION: Chronic pain is a critical limiting factor that delays rehabilitation after injury. Since currently available treatments are inadequate, the development of new therapeutics is a national healthcare priority. Years of animal experimentation have generated a multitude of mechanistic observations that explain the initiation and continuation of pain at a cellular and molecular level but new therapies have not emerged from this effort. Two key limitations are the nonspecific effects of typical small molecule drugs, and the inability to limit dangerous side effects caused by off-target actions when agents are delivered systemically. We have devised a solution that employs recombinant adeno-associated viral (AAV) vectors to express small analgesic peptides (called peptide aptamers) in sensory neurons following injection of the vector into the dorsal root ganglion (DRG). Substantial data show that injection at the level of the DRG is safe in animals and humans. This approach also employs only minute doses of agent, and restricts effects to the targeted peripheral sensory neurons of that segment. The peptide aptamers are designed to interfere with assembly, trafficking, or function of critical molecular targets that regulate peripheral sensory transduction and dorsal horn (DH) synaptic function, resulting in highly selective peripheral modulation of sensory neuron performance. This final feature is critical since genesis and perpetuation of chronic pain clearly involves plasticity in peripheral receptors and DH pain- transmitting synapses. The overall objective of this project is to define a novel strategy, AAV-delivered analgesic peptide aptamers, for modulating intracellular protein interactions of specific nociceptive pathways selectively in primary sensory neurons, and to test the efficacy of this approach as a molecular therapeutic tool for treating chronic pain. The proposed studies will test proof-of-concept on modulation of two critical pain signaling nodes, the N-type Ca2+ channel (CaV2.2) and the Transient Receptor Potential Vanilloid 1 (TRPV1) channel. Successful completion of the proposed research will set the stage for development of an entirely novel and functionally potent mode of segmental chronic pain therapy, and thereby convert the vast backlog of mechanistic discovery into practical treatments.