The Role of Ubiquitination in Methamphetamine Neurotoxicity (CDA)

描述（由申请人提供）：我的长期学术生涯目标是进行独立研究，发现影响多巴胺能系统的药物毒性的新分子途径，特别是甲基苯丙胺（MA），以便为其滥用者开发更好的治疗方法。泛素化在细胞过程中作为一个多功能信号而出现；因此，它是一种潜在的神经毒性新信号。没有关于高剂量MA后泛素化作用的数据；因此，我的直接目标是研究泛素化介导的E3连接酶（特别是帕金）在已知受MA毒性作用影响的蛋白质的调节中的作用，反之亦然，即泛素蛋白酶体系统，线粒体电子链蛋白，多巴胺转运蛋白（DAT）和囊泡单胺转运蛋白2 (VMAT2)，在MA毒性动物模型中。由于parkin可以抵抗多种细胞应激因子，包括影响线粒体功能的因子，因此计划确定parkin（或其他E3连接酶）是否可以在体内保护多巴胺能神经元抵抗MA。具体目的是：(1)研究parkin对MA神经毒性的影响，(2)研究MA对parkin功能和蛋白酶体催化性能的影响，(2)研究parkin功能下降与线粒体电子传递链活性之间的相互作用，(3)(4)研究MA后泛素化介导的DAT运输，(5)确定MA后DAT、parkin和VMAT2之间的分子联系。建议的指导研究将使我发展独立职业所需的技能，并学习新的技术，特别是获得扎实的MA毒性动物模型工作经验。这些知识和经验将有助于我的独立研究，以确定新的靶点，有望推动新药的开发，可以治疗或预防由MA引起的神经变性。由于涉及神经毒性的因素相互作用，了解它们的相互作用对于阐明药物干预的要点非常重要。到目前为止，还没有安全且经过测试的药物来治疗MA成瘾，急诊室医生需要使用新的MA解毒剂来治疗MA相关的过量。我的研究结果将有助于药物治疗的发展，以改善潜在的神经损伤和认知障碍，由于滥用MA。

DESCRIPTION (provided by applicant): My long-term academic career goal is to conduct independent research into the discovery of new molecular pathways involved in toxicity of drugs that affect dopaminergic system, particularly methamphetamine (MA), in order to develop better treatments for their abusers. Ubiquitination is emerging as a multifunctional signal in cellular processes; therefore, it is a potential novel signal in MA neurotoxicity. There is no data on the role of ubiquitination after high-dose MA; therefore, my immediate goal is to investigate the involvement of ubiquitination-mediating E3 ligases (particularly parkin) in regulation of proteins known to be affected by MA toxic actions, and vice versa, namely ubiquitin proteasomal system, mitochondrial electron chain proteins, dopamine transporter (DAT) and vesicular monoamine transporter 2 (VMAT2), in animal model of MA toxicity. Since parkin protects against a variety of cellular stressors, including agents affecting mitochondrial function, it is planned to determine whether parkin (or other E3 ligases) protects dopaminergic neurons against MA in vivo. The specific aims are: (1) to examine the effect of parkin on MA neurotoxicity, (2) to investigate the effect of MA on parkin function and the catalytic properties of the proteasome, (2) to investigate the interactions between decreased parkin function and mitochondrial electron transport chain activity, (3) (4) to investigate ubiquitination-mediated DAT trafficking after MA, and (5) to determine the molecular link between the DAT, parkin and VMAT2 after MA. The proposed mentored research will allow me to develop skills needed for independent career and to learn new techniques, particularly to acquire solid experience working with animal model of MA toxicity. Such knowledge and experience will help me with my independent studies directed toward the identification of new targets that may hopefully drive the development of novel drugs that can treat or prevent neurodegeneration caused by MA. Since factors involved in neurotoxicity interact with each other, it is very important to understand their interactions in order to elucidate points for pharmaceutical intervention. To date, there are no safe and tested medications for treating MA addiction and new MA antidotes for use by emergency room physicians to treat MA-related overdoses are needed. The results from my studies will assist the development of pharmacological therapies to ameliorate potential neuronal damage and cognitive impairments due to MA abuse.