Identification and functional characterization of ion channel-multiprotein complexes involved in nociception

慢性疼痛带来了巨大的挑战，因为目前的疼痛治疗无法控制。 此外，疼痛治疗剂靶向具有关键生理功能的分子，因此引起严重的副作用。鉴于这些困难，非常需要鉴定在初级伤害感受神经元中选择性表达或富集的分子和相关蛋白质支架。在这些神经元中，TRPA1和TRPV1通道作为有害刺激检测器，因此代表疼痛研究的主要目标。尽管对TRPA1和TRPV1进行了深入的研究，但对其蛋白质支架的组成知之甚少，特别是在疼痛状态下。本提案的目标是确定和研究多蛋白复合物与TRPA1和TRPV1通道的背景下，伤害。为此，该提案的重点是定性和定量鉴定蛋白质与内源性TRPA1和TRPV1通道在急性和炎症性疼痛状态下使用蛋白质组学方法。 将在TRPA1和TRPV1功能、伤害性信号传导和小鼠疼痛行为的背景下验证所鉴定的候选物的相关性。重要的是，关键参与介导或解偶联验证的蛋白质的结合TRPA1和TRPV1通道，分别域，应确定的最终目标，以调节各自的相互作用在体内，同时保持关键的生理功能完整。预计该项目有助于我们理解分子水平上的慢性疼痛过程，并提供新的分子靶点来特异性缓解慢性疼痛。

Chronic pain imposes substantial challenges, as it is unmanageable with current pain treatments. In addition, pain therapeutics target molecules with key physiological functions and therefore elicit severe side effects. In the light of these difficulties, identification of molecules and associated protein scaffolds, which are selectively expressed or enriched in primary nociceptive neurons, is highly desirable. In these neurons, TRPA1 and TRPV1 channels serve as noxious stimulus detectors and, therefore, represent primary targets for pain research. Despite intensive study of TRPA1 and TRPV1, little is known about the composition of their protein scaffold, especially during pain states. The goal of this proposal is to identify and study multiprotein complexes associated with TRPA1 and TRPV1 channels in the context of nociception. To this end, the proposal is focused on the qualitative and quantitative identification of proteins associated with endogenous TRPA1 and TRPV1 channels during states of acute and inflammatory pain using a proteomics approach. The relevance of identified candidates will be validated in the context of TRPA1 and TRPV1 function, nociceptive signaling and mouse pain behavior. Importantly, domains critically involved in mediating or uncoupling the binding of validated proteins to TRPA1 and TRPV1 channels, respectively, should be identified with the ultimate goal to modulate the respective interaction in vivo while leaving key physiological functions intact. It is anticipated that this project contributes to our understanding of the processes, which give way to chronic pain on the molecular level and provides new molecular targets to specifically relieve chronic pain conditions.