T-Channel Dysregulation during Alcohol Withdrawal: Mechanisms and Novel Therapies

戒酒期间 T 通道失调：机制和新疗法

• 批准号: 8316732
• 负责人: Melissa Ann Smaldino
• 金额: $ 4.22万
• 依托单位: WAKE FOREST UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-01 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8316732
• 关键词: Acute; Address; Alcohol Withdrawal Seizures; Alcohol abuse; Alcohol dependence; Alcohol withdrawal syndrome; Alcohols; Anticonvulsants; Blood - brain barrier anatomy; Brain; Calcium; Calcium Channel; Calcium Channel Blockers; Cells; Chronic; Dependence; Depressed mood; Development; Economic Burden; Effectiveness; Electrophysiology (science); Ethanol; Ethosuximide; Future; Generations; Hour; Individual; Interdisciplinary Study; Intervention; Ion Channel; Kindling (Neurology); Knowledge; Mediating; Membrane; Messenger RNA; Midline Thalamic Nuclei; Molecular; Monitor; Mus; Nervous system structure; Neuraxis; Neurons; Operative Surgical Procedures; Pharmacological Treatment; Phosphorylation; Play; Polymerase Chain Reaction; Post-Translational Protein Processing; Postdoctoral Fellow; Property; Protein Isoforms; Protein Kinase C; Proteins; Recovery; Relapse; Relative (related person); Research; Research Project Grants; Research Training; Reverse Transcription; Role; Seizures; Severities; Symptoms; T-Type Calcium Channels; Techniques; Testing; Thalamic structure; Therapeutic Intervention; Time; Training; Transcriptional Regulation; Western Blotting; Withdrawal; Withdrawal Symptom; Work; alcohol effect; alcohol exposure; ascorbate; attenuation; base; dehydroascorbate; implantation; insight; interdisciplinary approach; neural circuit; new therapeutic target; novel; patch clamp; pre-clinical; prevent; skills; therapeutic target; transcription factor REST

DESCRIPTION (provided by applicant): The primary objectives of this research training plan is to identify the molecular mechanisms responsible for T- type calcium channel (T-channel) dysregulation during alcohol withdrawal, and to determine whether T- channels can serve as preclinical therapeutic targets for alcohol withdrawal seizures. Withdrawal symptoms including seizures drive individuals to relapse, thus representing a significant barrier to recovery. With each successive withdrawal, symptoms increase in severity in a kindling-like phenomenon. Due to the depressing effects of alcohol in the CNS, compensatory mechanisms of progressive neuronal excitation ensue, with concurrent development of disrupted brain rhythms. Using an intermittent alcohol exposure paradigm, our lab has identified a disruption in the thalamic T-type calcium isoform, CaV3.2, during withdrawal that may underlie the generation and propagation of withdrawal seizures. The mechanisms responsible for the abnormal increases in excitability of T-channels are unknown, as well as whether or not pharmacological treatments targeting T-channels may be effective. I propose to address this potential mechanism at multiple levels. Specific Aim 1 will evaluate the use of ethosuximide, a non-specific T-channel antagonist, and the use of ascorbate, a specific CaV3.2 T-channel antagonist, as treatments against seizure acitivity during alcohol withdrawal. To further reveal mechanisms responsible for T-channel dysregulation during alcohol withdrawal, Aims 2 and 3 will evaluate if posttranslational modifications mediate the observed increase in excitability via protein kinase C (PKC) and the role of neuron-restrictive silencer factor (NRSF) in transcriptional regulation of CaV3.2 T-channel. This training plan proposed involves a multidisciplinary approach that will serve to advance the alcohol field by providing a better understanding of mechanisms and identifying novel targets for therapeutic intervention for individuals suffering from alcohol abus. This research approach will provide training in both electrophysiology and molecular techniques including surgical procedures, EEG analysis, patch clamp electrophysiology, quantitative reverse transcription polymerase chain reaction (qRT-PCR), and Western blot techniques. With guidance and support from the advisor and research associates, this project will provide the necessary training to pursue questions that will provide insight into the serious conditions that develop after chronic alcohol abuse. PUBLIC HEALTH RELEVANCE: Alcohol withdrawal symptoms, including seizures, drive individuals to relapse and represent a significant barrier to recovery. Indentifying mechanisms responsible and novel therapeutic targets is critical to help individuals recovering from alcohol abuse and will lower the economic burden and fatality rates. Investigating T-channel dysregulation during alcohol withdrawal will not only help us understand mechanisms for increased hyperexcitability but may be a promising strategy for future pharmacological interventions.

描述（由申请人提供）：本研究培训计划的主要目标是确定导致T-型钙通道（T通道）失调的分子机制，并确定T-通道是否可以用作临床前治疗靶标的酒精戒酒。 包括癫痫发作在内的戒断症状驱使个体复发，从而代表了恢复的重大障碍。 随着每次连续的戒断，在类似点燃的现象中症状的严重程度会增加。 由于酒精对中枢神经系统的影响下降，随着脑部节律破坏的同时发展，进行性神经元激发的补偿机制随之而来。 使用间歇性的酒精暴露范式，我们的实验室确定了丘脑T型钙亚型Cav3.2的破坏，这可能是戒断的产生和繁殖癫痫发作的基础。 导致T通道兴奋性异常增加的机制尚不清楚，以及针对T通道的药理治疗是否有效。 我建议在多个层面上解决这种潜在机制。 具体的目标1将评估非特异性T通道拮抗剂Ethosuximide的使用，并使用特定的Cav3.2 T通道拮抗剂Ascorbate用作抗饮酒期间癫痫发作阳性的治疗方法。 为了进一步揭示导致戒酒期间T通道功能失调的机制，AIMS 2和3将评估翻译后修饰是否介导了通过蛋白激酶C（PKC）观察到的兴奋性提高，而神经元反射式消音器（NRRSF）在转相调节中的作用 CAV3.2 T通道。 该培训计划涉及一种多学科的方法，该方法将通过更好地理解机制并确定针对患有酒精饮料的人的治疗干预措施的新目标来提高酒精领域的方法。 这种研究方法将提供有关电生理学和分子技术的培训，包括手术程序，脑电图分析，斑块夹电生理学，定量逆转录聚合酶链反应（QRT-PCR）和Western印迹技术。 在顾问和研究协会的指导和支持下，该项目将提供必要的培训，以提出问题，以洞悉长期酗酒后发展的严重状况。 公共卫生相关性：戒酒症状，包括癫痫发作，驱使个人复发并代表重大恢复的障碍。 负责任和新颖的治疗靶标的缩进机制对于帮助个人从酗酒中恢复并将降低经济负担和死亡率至关重要。 调查戒酒期间的T通道失调不仅会帮助我们了解过度兴奋性的机制，而且可能是未来药理干预措施的有前途的策略。