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对影响多巴胺能系统的药物，特别是甲基苯丙胺(MA)的毒性所涉及的新分子途径的发现进行独立研究，以开发更好的治疗方法。泛素化是细胞过程中的一种多功能信号，因此，它是MA神经毒性的一个潜在的新信号。目前还没有关于大剂量MA中毒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(尤其是Parkin)参与调节已知受MA毒性作用影响的蛋白质，反之亦然，在MA毒性的动物模型中，泛素蛋白酶体系统、线粒体电子链蛋白、多巴胺转运体(DAT)和囊泡单胺转运体2(VMAT2)。由于parkin可保护多种细胞应激源，包括影响线粒体功能的药物，因此计划在活体内确定parkin(或其他E3连接酶)是否保护多巴胺能神经元免受MA的影响。其具体目的是：(1)研究Parkin对MA神经毒性的影响，(2)MA对Parkin功能和蛋白酶体催化性质的影响，(2)研究Parkin功能降低与线粒体电子传递链活性的相互作用，(3)(4)研究MA后泛素化介导的DAT转运，(5)确定MA后DAT、Parkin和VMAT2之间的分子联系。拟议的指导性研究将使我能够发展独立职业所需的技能，并学习新技术，特别是获得与MA毒性动物模型一起工作的坚实经验。这些知识和经验将有助于我进行独立研究，以确定新的靶点，有望推动可以治疗或预防MA引起的神经变性的新药的开发。由于参与神经毒性的因素相互作用，了解它们之间的相互作用对于阐明药物干预的要点是非常重要的。到目前为止，还没有治疗MA成瘾的安全和经过测试的药物，需要新的MA解毒剂供急诊室医生使用来治疗MA相关的过量。我的研究结果将有助于药物治疗的发展，以改善由于MA滥用而造成的潜在神经元损伤和认知障碍。

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.