CYTOCHROME P450 2D6 VARIANTS IN NEUROTOXICITY

描述：(改编自申请者摘要)：本报告的目标 研究旨在阐明细胞色素P450 2D6变异体可能扮演的角色 神经毒性和神经退行性疾病。细胞色素基因的多态性 P450 2D6(P450 2D6)，一种已在人脑和 已知在外源和内源性的新陈代谢中起作用 前神经毒素被认为与帕金森氏症有关 疾病。我们的目标是推动对两国关系的理解 P450 2D6基因多态性与帕金森病在分子水平上的关系 最终，为复杂的相互作用提供机制基础 遗传和环境因素在这种疾病的病因中的作用。虽然 本申请侧重于对单个P450 2D6突变体的评估， 它表现出Arg296到Sys296突变，具有最高的风险因素 尚未确定为该疾病(=5.56)，制定的方案将是 一般适用于研究任何P450的作用 神经毒性变异(S)。具体地说，我们建议生产P450 2D6 及其变异体的数量足以进行体外研究 在COS和Sf9细胞中表达。我们将在体外定义 野生型和突变型P450 2D6的结构与功能关系 酶：(1)底物代谢和/或酶抑制； 以及(2)活性中间体或活性氧物种的产生 或一族结构上相关的四氢异喹啉，b-咔啉， 和吡啶类化合物，它们被确立或被提议有助于 帕金森氏症的病因学。最后，我们建议将一种 儿茶酚胺能细胞系大鼠嗜铬细胞瘤PC12细胞 类型和突变的酶。两者之间的差分敏感性 神经细胞应激和毒性标志物的转染体 我们体外鉴定的潜在神经原毒素的治疗 研究可能说明这种新陈代谢的细胞后果 不同之处。经过验证，这些细胞和类似的转基因神经元 细胞系可用于开发用于评估 内源性和外源性化合物的神经毒性 大脑代谢受P450 2D6的影响。

DESCRIPTION: (Adapted from Applicant's Abstract): The goal of this research is to elucidate the role that cytochrome P450 2D6 variants may play in neurotoxicity and neurodegenerative diseases. Polymorphism in cytochrome P450 2D6 (P450 2D6), an enzyme that has been identified in human brain and known to play a role in the metabolism of both foreign and endogenous pro-neurotoxins has been proposed to have an association with Parkinson's disease. It is our goal to move the understanding of the relationship between P450 2D6 polymorphism and Parkinson's disease to the molecular level and, ultimately, to provide a mechanistic basis for the complex interplay of genetic and environmental factors in the etiology of this disease. Although this application focuses on the evaluation of a single P450 2D6 mutant, which exhibits an Arg296 to Sys296 mutation and has the highest risk factor yet identified for the disease (=5.56), the protocol developed will be generally applicable to the investigation of the roles of any P450 variant(s) in neurotoxicity. Specifically, we propose to produce P450 2D6 and its variant in sufficient quantity for in vitro investigations by over expression in COS and Sf9 cells. We will define the in vitro structure-function relationships between the wild type and mutant P450 2D6 enzymes with regard to: (1) substrate metabolism and/or enzyme inhibition; and (2) the generation of reactive intermediates or reactive oxygen species or a family of structurally related tetrahydro-isoquinoline, b-carboline, and pyridine compounds, that are established or proposed to contribute to the etiology of Parkinson's disease. Finally, we propose to transfect a catecholaminergic cell line, rat pheochromocytoma PC12 cells, with the wild type and mutant enzymes. Differential sensitivities between the transfectants with regard to markers for neuronal stress and toxicity upon treatment with the potential pro-neurotoxins identified through our in vitro studies may illustrate the cellular consequences of such metabolic differences. With validation, these and analogously transfected neuronal cell lines may enable the development of assays for assessing the neurotoxicity of endogenous and foreign compounds that are responsible to cerebral metabolism by P450 2D6.