Estradiol and Zoster Associated Orofacial Pain

雌二醇和带状疱疹相关的口面部疼痛

基本信息

批准号：
10021211
负责人：
PHILLIP R KRAMER
金额：
$ 9.75万
依托单位：
TEXAS A&M UNIVERSITY HEALTH SCIENCE CTR
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：
2019-09-24 至 2020-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10021211
关键词：
Affect Area Aromatase Aromatase Inhibitors Attenuated Behavioral Assay Binding Biological Assay Biology Brain region Cell Count Cells Chronic Disease Clustered Regularly Interspaced Short Palindromic Repeats Complication DNA Diagnosis Diestrus Disease Electrophoretic Mobility Shift Assay Estradiol Estrogen Receptor alpha Estrogens FOS gene Female Gene Expression Genes Glutamate Decarboxylase Goals Gonadal Steroid Hormones Herpes zoster disease Herpesvirus Type 3 Human Hypersensitivity Immune Interneurons Lateral Luciferases Measurement Measures Molecular Mutate National Institute of Dental and Craniofacial Research Neural Inhibition Neurons Nociception Orofacial Pain Pain Pathogenesis Pathology Pathway interactions Patients Pharmaceutical Preparations Postherpetic neuralgia Production Proestrus Promoter Regions Rattus Reporter Reporting Reticular Cell Role Sex Differences Signal Transduction Site Skin Somatosensory Cortex Stains Structure of trigeminal ganglion Symptoms Technology Testing Thalamic structure Time Trigeminal Nuclei Vaccination Woman attenuation awake base chromatin immunoprecipitation chronic pain cingulate cortex experimental study gamma-Aminobutyric Acid inhibitory neuron insight knock-down male men midbrain central gray substance mutant nociceptive response novel orofacial pain model pain reduction promoter public health relevance reactivation from latency sex disparity transcription factor vesicular GABA transporter zona incerta

项目摘要

ABSTRACT People with herpes zoster (HZ) or “shingles” can suffer from orofacial pain. Orofacial HZ is caused by the reactivation of latent varicella zoster virus (VZV) in the trigeminal ganglia. Importantly, women report HZ pain three times more often than men, but the mechanism for this sex disparity is unknown. A screen by our lab of over 20,000 genes in five different regions of the orofacial pain pathway revealed that the greatest change in gene expression occurred in the thalamus, such that glutamate decarboxylase 2 (GAD2) and vesicular GABA transporter (VGAT) were elevated at proestrus (high 17 β-estradiol, E2) but not diestrus (low E2). In preliminary studies we showed that female rats have a greater VZV induced orofacial nociceptive response in comparison to males; mirroring the human sex difference. We also showed that reducing E2 production in male and female rats increases their nociceptive response and lastly, knock-down of VGAT expression in the thalamus increased the nociceptive response. Based on these studies we hypothesized that E2 attenuates orofacial nociception by increasing GAD2 or VGAT, enhancing GABAergic neural inhibition within the thalamus to reduce pain. To test this hypothesis, Aim 1 will characterize thalamic control of the VZV induced orofacial nociceptive response. The working hypothesis for Aim 1 is that GABA interneurons inhibit neurons in the thalamus to reduce VZV induced nociception. To test this hypothesis first, VZV induced nociception and neuronal activity will be recorded after attenuating inhibitory interneurons. Second, nociception and neuronal activity will be determined after modulating GAD2 and VGAT expression in these interneurons. Aim 2 will determine the role of sex steroids in modulating the VZV induced nociceptive response. Our working hypothesis is that E2 will increase expression of VGAT and GAD2 in the thalamus causing attenuation of neuronal activity and the nociceptive response. To test this hypothesis first, the nociceptive response and neuronal activity will be measured in female and male rats after administering E2 or reducing E2 production. Second, nociception and neuronal activity will be measured after knock-down of GAD2 and VGAT expression in rats with varied E2 levels. Aim 3 will characterize the mechanism by which sex steroids modulate VGAT and GAD2 to affect the VZV induced orofacial nociceptive response. Our working hypothesis is that E2 binds to estrogen receptor alpha (ERα) causing increased expression of VGAT and GAD2 in the thalamus resulting in attenuation of nociception. To test this idea, the nociceptive response and neuronal activity will be quantitated in rats after mutating ERα promoter sites using CRISPR/Cas9 technology. From these three aims we expect to determine 1) that GABA interneurons inhibit thalamic signaling to reduce orofacial pain, 2) that GAD2 and VGAT increases in these neurons through an ERα dependent mechanism, and 3) that GAD2 and VGAT are responsible, in part, for the sex difference observed in HZ pain. Identifying a mechanism by which E2 affects HZ-associated pain will provide new targets for treating people suffering from this chronic disorder.

抽象的带状疱疹 (HZ) 或“带状疱疹”患者可能会出现口面部疼痛。三叉神经节中潜伏的水痘带状疱疹病毒 (VZV) 重新激活重要的是，女性会出现带状疱疹疼痛。是男性的三倍，但这种性别差异的机制尚不清楚。口面部疼痛通路五个不同区域的 20,000 多个基因显示，最大的变化是基因表达发生在丘脑，例如谷氨酸脱羧酶 2 (GAD2) 和囊泡 GABA 转运蛋白 (VGAT) 在发情前期升高（高 17 β-雌二醇，E2），但在发情间期则不升高（低 E2）。初步研究表明，雌性大鼠在 VZV 诱导下的口面部伤害性反应更强与男性比较；反映了人类的性别差异。雄性和雌性大鼠的伤害性反应增加，最后，VGAT 表达被敲低根据这些研究，我们发现 E2 会减弱丘脑的伤害性反应。通过增加 GAD2 或 VGAT 来增强口面部伤害感受，增强 GABA 能神经抑制为了检验这一假设，目标 1 将表征丘脑对 VZV 诱导的控制。目标 1 的工作假设是 GABA 中间神经元抑制神经元。丘脑中减少 VZV 诱发的伤害感受首先检验这一假设，VZV 诱发伤害感受。减弱抑制性中间神经元后，将记录神经活动。调节这些中间神经元中的 GAD2 和 VGAT 表达后，将确定神经元活性。 2 将确定性类固醇在调节 VZV 诱导的伤害性反应中的作用。假设 E2 会增加丘脑中 VGAT 和 GAD2 的表达，从而导致神经活动和伤害性反应首先要检验这个假设，伤害性反应和在施用 E2 或减少 E2 产生后，将测量雌性和雄性大鼠的神经元活动。其次，敲除 GAD2 和 VGAT 表达后测量伤害感受和神经活动目标 3 将描述性类固醇调节 VGAT 的机制。和 GAD2 影响 VZV 诱导的口面部伤害性反应我们的工作假设是 E2 结合。雌激素受体 α (ERα) 导致丘脑中 VGAT 和 GAD2 表达增加为了测试这个想法，将伤害感受反应和神经活动。使用 CRISPR/Cas9 技术对大鼠中的 ERα 启动子位点进行突变后进行定量。我们期望确定 1) GABA 中间神经元抑制丘脑信号传导以减轻口面部疼痛，2) GAD2 和 VGAT 通过 ERα 依赖性机制在这些神经元中增加，并且 3) GAD2 和 VGAT 在一定程度上导致了带状疱疹疼痛的性别差异。 E2 影响带状疱疹相关的疼痛将为治疗患有这种慢性疾病的人提供新的靶点。