Pain Mechanisms in Fabry Disease

法布里病的疼痛机制

基本信息

批准号：
10381459
负责人：
Cheryl Louise Stucky
金额：
$ 76.95万
依托单位：
MEDICAL COLLEGE OF WISCONSIN
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-05-15 至 2023-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10381459
关键词：
3-Dimensional ANK1 gene Affect Afferent Neurons Age Alpha-galactosidase Analgesics Anatomy Animal Model Automobile Driving Behavioral Behavioral Assay Biology CRISPR/Cas technology Caliber Cells Chronic Consumption Cutaneous Data Disease Enzymes Exercise Exhibits FDA approved Fabry Disease Fatigue Female Fever Fiber Functional disorder Generations Genes Glycosphingolipids Goals Human Hypersensitivity Immune response Incidence Infusion procedures Intravenous infusion procedures Ion Channel Light Lipid Inclusion Lipids Lysosomal Storage Diseases Lysosomes Maintenance Mechanics Mediating Modeling Molecular Neonatal Screening Neurons Neuropathy Pain Pain Disorder Pain management Patients Peripheral Persistent pain Pharmacology Phenotype Physiological Piezo 2 ion channel Rat-1 Rattus Recombinants Risk Role Running Sensory Serum Site Spinal Ganglia Stimulus Stress System TRPA channel Tactile Testing Texture Therapeutic Intervention Time Touch sensation Transgenic Organisms allodynia base behavior test conditioned place preference disabling symptom enzyme replacement therapy gene replacement globotriaosylceramide hearing impairment improved in vivo knock-down male mechanical stimulus mechanotransduction mustard oil neuronal cell body novel pain behavior painful neuropathy preference receptor response small hairpin RNA somatosensory spontaneous pain treatment strategy

项目摘要

Project Summary The central goal of this project is to identify the cellular mechanisms that underlie the mechanical hypersensitivity and pain in Fabry Disease (FD), and thereby identify novel targets for improved pain treatment. FD is the most common lysosomal storage disease. It results from a deficiency of the lysosomal enzyme α- galactosidase A (α-Gal A) that leads to the accumulation of glycosphingolipids within the lysosomes of cells including dorsal root ganglia (DRG) neurons. Small fiber neuropathy is a hallmark of FD. Neuropathic-like pain begins around age 5, worsens with age, and affects 60-80% of all (male and female) patients. The pain is described as episodic “pain crises” triggered by fever, exercise, fatigue or stress, and chronic “permanent pain.” We created a transgenic rat model of FD using CRISPR/Cas9 to delete the gene encoding α-Gal A. Fabry rats closely recapitulate many phenotypes observed in patients, including elevated serum levels of α- galactosyl glycosphingolipids, spontaneous and mechanically-evoked pain behavior, pronounced lipid inclusions and aberrant accumulation of α-galactosyl glycosphingolipids in small-diameter DRG neurons and severe hearing loss. Sensory neurons somata from Fabry rats have sensitized rapidly adapting mechanical currents and sensitized responses to mustard oil, suggesting that Piezo2 and TRPA1 channel activities may be enhanced. Inhibition of TRPA1 alleviates the behavioral mechanical hypersensitivity in FD. Afferent fibers also show clear spontaneous activity, which may underlie the ongoing pain. The Scientific Premise of this proposal is that DRG sensory neurons in FD are dysfunctional and that the glycosphingolipids elevated in FD sensitize Piezo2 and TRPA1 channels in sensory neurons, resulting in the mechanical hypersensitivity and ongoing pain. This proposal will define the mechanisms that mediate Fabry disease pain in a top-down approach through Aims that 1) define the stimulus evoked, ongoing, and crisis-evoked pain behavior over time in male and female Fabry rats, 2) determine if the DRG is a key site for the generation and maintenance of mechanical and ongoing pain, and 3) interrogate whether Piezo2 and/or TRPA1 ion channels mediate the mechanical sensitization observed in the Fabry rat. In addition to identifying pain mechanisms that are FD- specific, these studies will have a broader impact by identifying mechanisms that could advance the understanding of mechanisms that underlie tactile allodynia and spontaneous pain in other types of neuropathic disorders and uncover novel roles for lipids in mechanotransduction mechanisms in somatosensory systems.

项目概要该项目的中心目标是确定机械作用背后的细胞机制法布里病（FD）中的过敏和疼痛，并由此确定改善疼痛治疗的新靶点。 FD 是最常见的溶酶体贮积病，由溶酶体酶 α- 缺乏引起。半乳糖苷酶 A (α-Gal A) 导致鞘糖脂在细胞溶酶体内积聚包括背根神经节 (DRG) 神经元，小纤维神经病变是 FD 的标志。 5 岁左右开始，随着年龄的增长而恶化，影响 60-80% 的患者（男性和女性）。被描述为由发烧、运动、疲劳或压力引发的间歇性“疼痛危机”，以及慢性“永久性疼痛” 我们使用 CRISPR/Cas9 删除编码 α-Gal A 的基因，创建了 FD 转基因大鼠模型。 Fabry 大鼠密切再现了在患者中观察到的许多表型，包括血清 α-水平升高半乳糖鞘糖脂，自发性和机械诱发的疼痛行为，明显的脂质小直径 DRG 神经元中 α-半乳糖鞘糖脂的内含物和异常积累法布里大鼠的感觉神经元体细胞已经变得敏感，能够快速适应机械。电流和对芥子油的敏感反应，表明 Piezo2 和 TRPA1 通道活动可能是 TRPA1 的抑制也减轻了 FD 的行为机械过敏。显示出明显的自发活动，这可能是持续疼痛的基础。建议认为 FD 中的 DRG 感觉神经元功能失调，并且 FD 中的糖鞘脂升高使感觉神经元中的 Piezo2 和 TRPA1 通道敏感，导致机械超敏反应该提案将定义自上而下介导法布里病疼痛的机制。该方法的目标是 1) 定义随着时间的推移刺激诱发的、持续的和危机诱发的疼痛行为在雄性和雌性法布里大鼠中，2) 确定 DRG 是否是产生和维持机械性和持续性疼痛，3) 询问 Piezo2 和/或 TRPA1 离子通道是否介导除了确定 FD- 的疼痛机制外，还观察到在法布里大鼠中观察到的机械敏化。具体而言，这些研究将通过确定可促进发展的机制而产生更广泛的影响。了解其他类型的触觉异常性疼痛和自发性疼痛的机制神经性疾病并揭示脂质在机械传导机制中的新作用体感系统。