14-3-3 gamma regulation of peripheral kappa opioid receptor function

摘要 近三分之一的美国人口患有慢性疼痛。这种痛苦会导致 重大活动限制在近1100万人。阿片类药物被用来治疗疼痛，但有很多 负面影响，如呼吸抑制、上瘾和镇静。因为这些负面影响是 在中枢神经系统的调节下，安全有效地治疗疼痛的一个有希望的方法是 瞄准周围神经系统。信号由外周神经系统发送到大脑，并 被解释为痛苦。通过专门抑制这些信号的传输，疼痛可以得到缓解。这个 局部麻醉剂的显著止痛效果说明了这种方法的价值。 外周受限的阿片类药物可以靶向外周神经元，克服局部麻醉药的局限性， 比如运动神经元抑制。然而，外周的阿片受体受到独特的调节，而且效果不佳。 已知的长期抑制机制会降低外周受限阿片类药物的有效性。这 研究已有初步证据表明，调节分子14-3-3γ介导了对 通过kappa阿片受体(KOR)发挥抗伤害作用。此外，14-3-3γ似乎偏向KOR信号 走向促伤害性丝裂原活化蛋白激酶(MAPK)通路。这项奖学金将：(1) 建立14-3-3γ与KOR直接相互作用，(2)鉴定14-3-3γ介导的KOR信号通路， (3)评价KOR伤害性信号在体内的变化。这些拟议的研究将阐明 14-3-3-γ在调节KOR介导的外周感觉神经元抗伤害感受中的作用。因此，这些 研究将对安全有效的外周限制阿片类药物的使用产生重大影响 疼痛的治疗。 此外，这些研究将作为申请者的杰出培训工具。创新的 研究方法结合了异源表达系统、大鼠原代神经元培养和行为学 伤害性测试，这将确保在药理学，细胞信号， 以及啮齿动物的行为。在申请者博士后咨询委员会的指导下，培训目标 开发新技术，扩大对新领域的理解，完善沟通，以及 将实现科研产品的生产。最终，这一奖学金将为成功的 作为一名研究人类疾病中阿片受体信号的独立研究员。

ABSTRACT Nearly one third of the population of the United States suffers from chronic pain. This pain causes major activity restrictions in nearly 11 million people. Opioids are used for the treatment of pain, but have many negative effects such as respiratory depression, addiction, and sedation. As these negative effects are mediated by the central nervous system, a promising approach for the safe and effective treatment of pain is to target the peripheral nervous system. Signals are sent by the peripheral nervous system to the brain and are interpreted as pain. By specifically inhibiting the transmission of these signals, pain can be alleviated. The value of this approach is illustrated by the pronounced analgesic effectiveness of local anesthetics. Peripherally restricted opioids can target peripheral neurons and overcome the limitations of local anesthetics, such as motor neuron inhibition. However, opioid receptors in the periphery are uniquely regulated, and poorly understood mechanisms of long-term inhibition reduce the effectiveness of peripherally restricted opioids. This study has preliminary evidence that the regulatory molecule 14-3-3γ mediates long-term inhibition of antinociception through the kappa opioid receptor (KOR). Further, 14-3-3γ appears to bias KOR signaling towards the pronociceptive mitogen-activated protein kinase (MAPK) pathways. This fellowship will: (1) establish that 14-3-3γ directly interacts with KOR, (2) identify KOR signaling pathways mediated by 14-3-3γ, and (3) evaluate the changes in KOR nociceptive signaling in vivo. These proposed studies will elucidate the role of 14-3-3γ in regulating KOR-mediated antinociception in peripheral sensory neurons. As a result, these studies will have significant impact on the use of peripherally restricted opioids for the safe and effective treatment of pain. Additionally, these studies will serve as an outstanding training vehicle for the applicant. The innovative research approach combines a heterologous expression system, rat primary neuronal culture, and behavioral nociception assays, which will ensure comprehensive training in the fields of pharmacology, cellular signaling, and rodent behavior. With the guidance of the applicant’s postdoctoral advisory committee, the training goals of developing new techniques, expanding understanding in a new field, refining communications, and producing research products will be realized. Ultimately, this fellowship will lay the foundation for a successful career as an independent investigator studying opioid receptor signaling in human disease.