PROTEASOME AND PARKIN AS DRUG TARGETS AGAINST METHAMPHETAMINE TOXICITY

蛋白酶体和 Parkin 作为对抗甲基苯丙胺毒性的药物靶标

• 批准号: 8578758
• 负责人: Anna Moszczynska
• 金额: $ 32.3万
• 依托单位: WAYNE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-15 至 2018-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8578758
• 关键词: 26S proteasome; Acute; Animals; Attenuated; Behavior; Blood - brain barrier anatomy; Brain; Brain Diseases; Cell Death; Cell Survival; Chronic; Clinical; Confocal Microscopy; Corpus striatum structure; Data; Development; Dissociation; Dose; Drug Targeting; Etiology; Evaluation; Exposure to; Goals; HIV-1; Human; Immunohistochemistry; Impaired cognition; Impairment; In Vitro; Injection of therapeutic agent; Intervention; Knock-out; Lead; Link; Mediating; Methamphetamine; Methods; Molecular; Neurons; Overdose; Oxidative Stress; Parkinson Disease; Peptides; Pharmaceutical Preparations; Pharmacologic Substance; Process; Proteins; Rattus; Recovery; Relative (related person); Research; Role; System; Therapeutic; Toxic effect; Ubiquitin; Up-Regulation; Variant; base; combat; design; dopaminergic neuron; gamma-Aminobutyric Acid; in vivo; inhibitor/antagonist; methamphetamine abuse; multicatalytic endopeptidase complex; neuronal survival; neuroprotection; neurotoxic; neurotoxicity; novel; novel strategies; overexpression; parkin gene/protein; prevent; protein aggregate; psychostimulant; public health relevance; synuclein; ubiquitin-protein ligase

DESCRIPTION (provided by applicant): Methamphetamine (METH) is a highly addictive psychostimulant drug that is neurotoxic when taken at high doses chronically or acutely. METH selectively damages striatal dopaminergic terminals in experimental animals and humans. Despite years of active research on METH neurotoxicity, no specific medications have been developed to counteract the damaging effects that METH has on the brain. Due to its widespread abuse, there is a compelling need for effective pharmaceuticals that can protect and/or restore the brain from the toxic effects of acute METH overdose and chronic METH abuse. Thus, it is necessary to identify molecular drug targets in order to develop novel pharmaceuticals. My long-term goal is to develop neuroprotective therapies to treat the toxic effects of METH use. The goal of the proposed research is to better understand the molecular mechanisms regulating the ubiquitin-proteasome system in the METH-exposed rat brain and to determine whether two components of this system, proteasome and the E3 ligase parkin, are potential pharmaceutical targets that can be used to promote [survival and recovery] of dopaminergic terminals in vivo after toxic doses of [binge and chronic] METH. Both proteasome and parkin are decreased shortly after binge METH administration, and those deficits are also involved in the etiology of Parkinson's disease. We hypothesize that increasing their functions will provide neuroprotection in rats chronically and acutely exposed to METH. Specific Aim 1 will evaluate the relative roles of 20S and 26S proteasomes on the [survival of dopaminergic terminals and their recovery from binge and chronic] METH by using proteasomal inhibitors and a novel approach to regulate proteasomal activity, namely, systemic injections of TAT-tagged peptides that interfere with proteasomal assembly. TAT is a domain of the human immunodeficiency virus type 1 that rapidly crosses the blood brain barrier. These two forms of the proteasome behave differently upon exposure to METH- induced oxidative stress; thus, evaluation of their respective roles in METH neurotoxicity is warranted. Specific Aim 2 will evaluate the role of parkin in the [survival of and recovery from binge and chronic] METH using wild- type, parkin-overexpressing, and parkin knock-out rats. The role of parkin in the formation of intracellular inclusions in the nigrostriatal dopamine neurons will be investigated using immunohistochemistry and confocal microscopy. Specific Aim 3 will determine whether proteasomes and parkin are functionally linked in the METH-exposed rat brain. For clinical intervention purposes, it is important to know how variations in parkin levels influence 20S and 26S function and vice versa. These aims are conceptually linked as they investigate regulatory processes within the ubiquitin-proteasome system that may be important for [endogenous survival and recovery mechanisms] in dopamine neurons and, therefore, clinically important. The findings from the proposed research may lead to novel treatments for METH users.

描述（由申请人提供）：甲基苯丙胺（METH）是一种高度成瘾的精神兴奋药物，长期或急性服用高剂量时会产生神经毒性。冰毒选择性地损害实验动物和人类的纹状体多巴胺能末端。尽管对冰毒神经毒性进行了多年的积极研究，但尚未开发出特定药物来抵消冰毒对大脑的损害作用。由于其广泛滥用，迫切需要有效的药物来保护和/或恢复大脑免受急性冰毒过量和慢性冰毒滥用的毒性作用。因此，有必要确定分子药物靶点以开发新药物。我的长期目标是开发神经保护疗法来治疗冰毒使用的毒性作用。本研究的目的是更好地了解 METH 暴露大鼠大脑中调节泛素-蛋白酶体系统的分子机制，并确定该系统的两个组成部分——蛋白酶体和 E3 连接酶 Parkin——是否是潜在的药物靶标。用于在中毒剂量的[暴饮暴食和慢性]冰毒后促进体内多巴胺能末端的[生存和恢复]。在大量服用冰毒后不久，蛋白酶体和帕金蛋白都会减少，这些缺陷也与帕金森病的病因有关。我们假设增加它们的功能将为长期和急性暴露于冰毒的大鼠提供神经保护。具体目标 1 将通过使用蛋白酶体抑制剂和调节蛋白酶体活性的新方法（即全身注射 TAT 标记）来评估 20S 和 26S 蛋白酶体对 [多巴胺能末端的存活及其从暴食和慢性中恢复] METH 的相对作用干扰蛋白酶体组装的肽。 TAT 是 1 型人类免疫缺陷病毒的一个结构域，可快速穿过血脑屏障。这两种形式的蛋白酶体在暴露于冰毒诱导的氧化应激时表现不同。因此，有必要评估它们各自在冰毒神经毒性中的作用。具体目标 2 将使用野生型、parkin 过度表达和 Parkin 敲除大鼠评估 Parkin 在[暴食和慢性]冰毒的存活和恢复中的作用。将使用免疫组织化学和共聚焦显微镜研究parkin在黑质纹状体多巴胺神经元细胞内包涵体形成中的作用。具体目标 3 将确定蛋白酶体和 Parkin 在暴露于冰毒的大鼠大脑中是否存在功能联系。出于临床干预目的，了解 Parkin 水平的变化如何影响 20S 和 26S 功能非常重要，反之亦然。这些目标在概念上是相互联系的，因为它们研究泛素蛋白酶体系统内的调节过程，这可能对多巴胺神经元的[内源性存活和恢复机制]很重要，因此具有临床重要性。拟议研究的结果可能会为冰毒使用者带来新的治疗方法。