Characterization of the BMP signaling pathways that produce nociceptor sensitization in Drosophila

果蝇中产生伤害感受器敏化的 BMP 信号通路的表征

• 批准号: 9022858
• 负责人: GEOFFREY GANTER
• 金额: $ 44.35万
• 依托单位: UNIVERSITY OF NEW ENGLAND
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-30 至 2019-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9022858
• 关键词: Address; Adverse effects; Affect; Alleles; American; Analgesics; BMP2 gene; Biological Assay; Bone Morphogenetic Proteins; Cell Nucleus; Cell Separation; Cell membrane; Cells; Development; Dose; Drosophila genus; Drosophila sax protein; Emotional; Epidermis; Exhibits; Future; Gene Expression; Gene Targeting; Genes; Genetic; Genetic Models; Genetic Techniques; Genome; Healed; Health; Human; Injury; Investigation; Knowledge; Larva; Life; Ligands; Mammals; Measures; Modeling; Neurons; Nociception; Nociceptors; Opioid Analgesics; Orthologous Gene; Output; Pain; Pain management; Pathway interactions; Pharmaceutical Preparations; Procedures; Process; Protein Family; Quality of life; RNA Interference; RNA Sequences; Receptor Cell; Role; Signal Pathway; Signal Transduction; Signaling Protein; Stimulus; System; Testing; Tissues; Translating; UV induced; Ultraviolet Rays; Up-Regulation; Wit; addiction; allodynia; avoidance behavior; bone morphogenetic protein receptors; chronic pain; economic impact; extracellular; fly; healing; human disease; improved; interest; magnetic beads; member; mutant; novel; overexpression; prevent; public health relevance; receptor; research study; tool; transcriptome sequencing; ultraviolet; ultraviolet damage

 DESCRIPTION (provided by applicant): Normal pain promotes health by warning us of potential tissue damage, but abnormal pain compromises the quality of life for millions around the world. The impact of pain on the US economy has been estimated at over $600B. Pain sensitization, an increase in pain signaling after tissue damage, also promotes health by preventing re-injury during healing. Some types of abnormal pain, including types of 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 accentuated with 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 epidermis of the fly larva is damaged by a controlled dose of ultraviolet (UV) light, the fly exhibits allodynia, a type of nociceptive sensitization. Ths means that the damaged fly will react, with a nocifensive avoidance behavior in the form of an unmistakable corkscrew roll, to a thermal stimulus that most undamaged flies find innocuous. This damage induced sensitization paradigm has previously been used to demonstrate that the nociceptor neuron requires signaling by the Bone Morphogenetic Proteins (BMP) pathway in order to produce allodynia. Many BMP signaling components in the fly are very similar to their mammalian orthologs. Our preliminary results show that one of the three known BMPs in the fly, dpp, and its receptor put are necessary in the nociceptor neurons for allodynia following UV damage. Aim 1 will examine the necessity of the other BMP ligands gbb and scw, and the other BMP receptors tkv, sax, wit and babo. This will be accomplished by targeting an RNAi silencing allele of each gene specifically to the nociceptor neurons by the Gal4/UAS system. Aim 2 will examine the intracellular relay components mad, med, smox, R-SMAD, shn and brk, again using Gal4/UAS and RNAi tools. Aim 3 will describe the transcriptional output of the BMP signaling pathway in the nociceptor neuron by magnetic bead- assisted cell sorting and RNA sequencing. This procedure will identify the target genes regulated by the pathway in the nociceptor neurons. Finally, the necessity for the expression of these target genes will be tested by analyzing RNAi mutants in the UV sensitization paradigm. The proposed studies will identify the components of the BMP signaling pathway that are required for allodynia. Because of the high degree of functional conservation between fly and mammalian BMPs, components identified by these experiments represent targets for novel medications for the treatment of abnormal pain in humans.

 描述（由申请人提供）：正常疼痛通过警告我们潜在的组织损伤来促进健康，但异常疼痛损害了世界各地数百万人的生活质量。痛苦对美国经济的影响估计超过6000亿美元。疼痛敏化，即组织损伤后疼痛信号的增加，也通过防止愈合期间的再损伤来促进健康。某些类型的异常疼痛，包括慢性疼痛，是由疼痛敏感系统的失调引起的。现有的慢性疼痛治疗是不够的，部分原因是我们最好的止痛药，阿片类药物的有害副作用，随着使用时间的延长而加重。我们迫切需要更好的治疗异常疼痛的方法。我们建议通过利用果蝇模型的强大遗传工具包来揭示止痛药的新靶点。当果蝇幼虫的表皮被受控剂量的紫外线（UV）光损伤时，果蝇表现出异常性疼痛，这是一种伤害性敏化。这意味着受损的苍蝇会对大多数未受损的苍蝇认为无害的热刺激做出反应，表现为明显的螺旋形滚动形式的伤害性回避行为。这种损伤诱导的致敏范例先前已被用于证明伤害感受器神经元需要通过骨形态发生蛋白（BMP）途径的信号传导以产生异常性疼痛。苍蝇中的许多BMP信号成分与它们的哺乳动物直系同源物非常相似。我们的初步研究结果表明，三个已知的BMP在苍蝇，DPP，和它的受体放在伤害性感受器神经元的异常性疼痛后，紫外线损伤是必要的。目的1将检查其他BMP配体gbb和scw以及其他BMP受体tbb、sax、wit和babo的必要性。这将通过Gal 4/UAS系统将每个基因的RNAi沉默等位基因特异性靶向伤害感受器神经元来实现。目的2将再次使用Gal 4/UAS和RNAi工具检查细胞内中继组件mad、med、smox、R-SMAD、shn和brk。目的3将描述通过磁珠辅助细胞分选和RNA测序的BMP信号通路在伤害感受器神经元中的转录输出。这一过程将确定伤害感受器神经元中受该通路调控的靶基因。最后，这些靶基因的表达的必要性将通过分析RNAi突变体中的UV敏化范例进行测试。拟议的研究将确定异常性疼痛所需的BMP信号通路的组成部分。由于果蝇和哺乳动物BMP之间的高度功能保守性，这些实验鉴定的组分代表了用于治疗人类异常疼痛的新型药物的靶点。