Characterization of the signaling pathways that produce nociceptor sensitization in Drosophila

果蝇中产生伤害感受器敏化的信号通路的表征

基本信息

批准号：
10046777
负责人：
GEOFFREY GANTER
金额：
$ 42.6万
依托单位：
UNIVERSITY OF NEW ENGLAND
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-09-30 至 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10046777
关键词：
Address Affect Afferent Neurons Affinity Chromatography Analgesics Animals Behavioral Bioinformatics Biological Models Bone Morphogenetic Proteins C-terminal binding protein Cornea Coupled Data Data Set Dose Drosophila genus Emotional Epidemic Exhibits Future Gene Targeting Genes Genetic Genetic Models Genetic Transcription Genome Goals Health Promotion Histologic Human Hypersensitivity Injury Intractable Pain Investigation Knowledge Larva Lead Life Ligands Light Mammals Maps Measures Mediating Messenger RNA Modeling Neurons Nociception Nociceptors Opiate Addiction Opioid Orthologous Gene Pain Pain management Pathologic Pathway interactions Peripheral Pharmaceutical Preparations Phosphorylation Population Process Quality of life RNA RNA Interference Rattus Recovery Ribosomes Rodent Role Signal Pathway Signal Transduction Signaling Protein Stimulus System Testing Therapeutic Time Tissues Transcriptional Regulation Transducers Translating Translations UV induced UV injury Ultraviolet Rays Wit addiction allodynia avoidance behavior chronic pain chronic painful condition drug efficacy economic impact experimental study extracellular fly healing human disease improved injured knock-down neurophysiology new therapeutic target non-opioid analgesic novel pain reduction receptor response response to injury side effect tool transcription factor transcriptome sequencing ultraviolet

项目摘要

Normal pain promotes health by warning us of potential tissue damage, but abnormal pain reduces the quality of life for millions around the world. Pain sensitization after injury also promotes health by reducing re-injury during healing. Some types of abnormal pain, including chronic pain, result from dysregulation of the pain sensitization system. Available treatment for chronic pain is inadequate, in part because the deleterious side effects of our best analgesics, the opioids, are made more hazardous by longer use. Better treatments for abnormal pain are badly needed. We propose to reveal novel targets for pain medications by exploiting the powerful genetic toolkit of the Drosophila model. When the fruit fly larva is injured by a controlled dose of ultraviolet (UV) radiation, the animal exhibits allodynia, a manifestation of nociceptive sensitization. This means that the injured animal will react with a nocifensive avoidance behavior in the form of an unmistakable corkscrew roll, in response to a thermal stimulus that most uninjured animals find innocuous. This injury induced sensitization paradigm has previously been used to demonstrate that the nociceptor neuron requires signaling by the Bone Morphogenetic Proteins (BMP) pathway to produce allodynia. BMP signaling components in the fly are very similar to their mammalian orthologs. Preliminary results indicate that a transcriptional and translational response is necessary in the nociceptor neurons in order to produce allodynia following injury. Aim 1 will examine the necessity of a set of BMP related transcriptional regulators like brinker, yorkie and bantam. This will be accomplished by targeting an RNAi silencing construct of each gene specifically to the nociceptor neurons using the Gal4/UAS system. Aim 2 will bring additional BMP pathway and other components to light by identifying transcriptional and translationally regulated genes, again using Gal4/UAS gene targeting, ribosome-tagging and immunopurification tools, RNA sequencing and bioinformatics. Aim 3 will begin to translate the discoveries made in Drosophila into mammalian systems by testing BMP pathway inhibiting drugs for efficacy in providing relief from injury-induced pain sensitization. The proposed studies have the potential to identify all components regulated by transcription and translation in the signaling pathways required for allodynia. Because of the high degree of functional conservation between fly and mammalian BMPs, components identified by these experiments may represent targets for novel medications for the treatment of abnormal pain in humans.

正常疼痛通过警告我们潜在的组织损害来促进健康，但异常疼痛降低了质量全世界数百万人的生活。受伤后的疼痛敏化还可以通过减少伤害来促进健康在康复期间。疼痛失调引起的某些类型的异常疼痛，包括慢性疼痛致敏系统。可用的慢性疼痛治疗不足，部分原因是有害的一面我们最好的镇痛药的影响，阿片类药物，通过更长的使用使危险更大。更好的治疗方法急需异常的疼痛。我们建议通过利用止痛药的新目标果蝇模型的强大遗传工具包。当果蝇幼虫受到受控剂量伤害时紫外线（紫外线）辐射，动物表现出异肌，是伤害感受敏化的表现。这这意味着受伤的动物将以明确的形式以一种单一的回避行为做出反应开瓶器卷，响应大多数未受伤的动物发现无害的热刺激。这种伤害诱导的敏化范式以前已被用来证明伤害感受器神经元需要骨形态发生蛋白（BMP）途径的信号传导产生异常性疾病。 BMP信号飞行中的成分与它们的哺乳动物直系同源物非常相似。初步结果表明在伤害感受器神经元中必须进行转录和翻译反应，以产生异常性症受伤后。 AIM 1将检查一组与BRINKER（如Brinker）相关的转录调节器的必要性，约克和矮脚鸡。这将通过靶向每个基因的RNAi沉默构建专门用于使用GAL4/UAS系统的伤害感受器神经元。 AIM 2将带来额外的BMP途径以及通过识别转录和翻译调节基因的其他组件，再次使用 GAL4/UAS基因靶向，核糖体标记和免疫力治疗工具，RNA测序和生物信息学。 AIM 3将开始将果蝇中的发现转化为哺乳动物系统测试BMP途径抑制药物在减轻损伤引起的疼痛敏感性方面的功效。这拟议的研究有可能识别由转录和翻译调节的所有组件异常性ni症所需的信号通路。由于苍蝇之间的功能保护很高和哺乳动物BMP，这些实验确定的成分可能代表新颖的目标治疗人类异常疼痛的药物。