A Mechanistic Study of Methamphetamine Neurotoxicity: Involvement of HO-1 & MnSO

甲基苯丙胺神经毒性的机制研究：HO-1 的参与

• 批准号: 7541182
• 负责人: Melinda Lea Asbury
• 金额: $ 3.53万
• 依托单位: MARSHALL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-01 至 2011-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7541182
• 关键词: Acute; Antioxidants; Apoptosis; Apoptotic; Attenuated; Biological Models; Brain; Brain Injuries; Cell Death; Cell Line; Cell model; Cells; Communities; Dementia; Development; Disease; Dopamine; Dopamine D1 Receptor; Dopamine Receptor; Epidemic; Generations; Goals; Heme; Human; Impaired cognition; In Vitro; Isoenzymes; Light; Lipid Peroxidation; Literature; Manganese Superoxide Dismutase; Mediating; Methamphetamine; Methods; Mitochondria; Modeling; Molecular; Nerve Degeneration; Nervous System Trauma; Neurodegenerative Disorders; Neuroepithelioma; Neurons; Nitrates; Nitric Oxide Synthase; Nitrogen; Oxygen; Oxygenases; Patients; Peroxonitrite; Pharmaceutical Preparations; Process; Production; Property; Proteins; Reactive Nitrogen Species; Research; Research Proposals; Scheme; Schizophrenia; Small Interfering RNA; Superoxides; Supplementation; Testing; Therapeutic; Therapeutic Intervention; Toxic effect; Tyrosine; United States; Up-Regulation; addiction; area striata; brain cell; clinically relevant; combat; dopamine transporter; effective therapy; experience; feeding; heme oxygenase-1; in vitro Model; innovation; knock-down; mesoporphyrin IX; mimetics; neurotoxicity; nitration; novel; novel therapeutic intervention; postsynaptic; public health relevance; response; stressor

DESCRIPTION (provided by applicant): Project Summary: The goal of this research is to help bridge the gap between methamphetamine (MA) neurotoxicity and effective therapeutic intervention. Using a D1 dopamine (DA) receptor-expressing neuroepithelioma cell line (SK-N-MC) that lacks DA transporters, we have developed an in vitro system that models post-synaptic neurons directly targeted by MA toxicity. By utilizing this model we will study novel molecular mechanisms involved in MA-induced neurotoxicity, in particular the contribution of heme oxygenase-1 (HO-1), reactive nitrogen species (RNS) and manganese superoxide dismutase (MnSOD). We hypothesize that DA stimulation, via a process mediated by HO-1, leads to generation of RNS and mitochondrial-mediated apoptosis through nitration of MnSOD. Our hypothesis will be tested in the following three specific aims: 1) Test whether HO-1 expression is involved in RNS production, 2) Test whether inhibition of HO-1 attenuates DA-induced mitochondrial-mediated apoptosis, and 3) Determine if MnSOD is nitrated and if treatment with a general antioxidant or mimetic attenuates DA-induced nitration and/or mitochondrial-mediated apoptosis. We will investigate ways to decrease MA neurotoxicity through studies aimed at manipulation of HO-1 and MnSOD activity in order to limit RNS production and subsequent apoptosis. This will be done using innovative methods such as siRNA-mediated knock-down of HO-1 expression, or increased MnSOD catalytic activity via pharmacological mimetics as well as utilizing compounds that have previously demonstrated clinical relevance in other disease states, such as Sn (IV) mesoporphyrin IX. These findings will enable the development of new therapeutic approaches that, when given as a pre-treatment, will minimize further neurological damage to those struggling with the disease of MA addiction. PUBLIC HEALTH RELEVANCE: Methamphetamine is a particularly devastating drug as its abuse has been shown to cause localized brain damage equivalent to that seen in patients with early dementia and greater than that seen in those with schizophrenia. In order to develop effective therapeutics to combat the brain damage seen in methamphetamine addicts, it is essential to first understand how the brain damage occurs. This research proposal is aimed at determining, on a molecular level, what happens inside brain cells to cause them to die after being exposed to methamphetamine. In light of this new-found information, the scientific community will be closer to developing new and effective treatments for methamphetamine toxicity.

描述（由申请人提供）： 项目摘要：本研究的目标是帮助弥合甲基苯丙胺 (MA) 神经毒性和有效治疗干预之间的差距。使用缺乏 DA 转运蛋白的表达 D1 多巴胺 (DA) 受体的神经上皮瘤细胞系 (SK-N-MC)，我们开发了一种体外系统，可以模拟 MA 毒性直接靶向的突触后神经元。通过利用该模型，我们将研究 MA 诱导的神经毒性的新分子机制，特别是血红素加氧酶-1 (HO-1)、活性氮 (RNS) 和锰超氧化物歧化酶 (MnSOD) 的贡献。我们假设 DA 刺激通过 HO-1 介导的过程导致 RNS 的产生，并通过 MnSOD 的硝化导致线粒体介导的细胞凋亡。我们的假设将在以下三个具体目标中得到检验：1) 测试 HO-1 表达是否参与 RNS 产生，2) 测试 HO-1 的抑制是否会减弱 DA 诱导的线粒体介导的细胞凋亡，以及 3) 确定 MnSOD 是否被硝化，以及用一般抗氧化剂或模拟物处理是否会减弱 DA 诱导的硝化和/或线粒体介导的硝化 细胞凋亡。我们将通过旨在操纵 HO-1 和 MnSOD 活性的研究来研究降低 MA 神经毒性的方法，以限制 RNS 的产生和随后的细胞凋亡。这将通过创新方法来完成，例如 siRNA 介导的 HO-1 表达敲低，或通过药理学模拟物增加 MnSOD 催化活性，以及​​利用先前已证明与其他疾病状态具有临床相关性的化合物，例如 Sn (IV) 中卟啉 IX。这些发现将有助于开发新的治疗方法，当作为预处理时，将最大限度地减少对 MA 成瘾患者的进一步神经损伤。 公共卫生相关性：甲基苯丙胺是一种破坏性特别大的药物，因为滥用它会造成局部脑损伤，其程度相当于早期痴呆症患者的程度，并且比精神分裂症患者的程度更大。为了开发有效的疗法来对抗甲基苯丙胺成瘾者的脑损伤，首先必须了解脑损伤是如何发生的。这项研究计划的目的是在分子水平上确定脑细胞在接触甲基苯丙胺后会发生什么导致它们死亡。根据这一新发现的信息，科学界将更接近开发新的有效治疗甲基苯丙胺毒性的方法。