Regulation and Consequences of Cytochrome P450 2E1

项目摘要 细胞色素P450 2 E1（CYP 2 E1）是一种独特的、高度保守的、高度调控的哺乳动物P450 单加氧酶CYP 2 E1经历广泛的转录后和翻译后调节，包括 双峰靶向内质网和线粒体。没有已知的功能丧失突变， 人类群体，并且没有已知的改变蛋白质编码区的多态性变体， 表明CYP 2 E1的重要内源性作用。然而，在啮齿类动物模型中敲除CYP 2 E1并没有 产生了引人注目的表型，并且据报道对饮食诱导的肥胖有保护作用， 非酒精性脂肪肝和乙醇诱导的毒性。CYP 2 E1的治疗靶向已经被 建议用于酒精性肝损伤后的肝脏修复，并提高生酮饮食的功效 癫痫的治疗然而，在我们对植物的内源性功能的理解上， CYP 2 E1在人体中，特别是在长期使用中，安全抑制CYP 2 E1之前必须解决的问题。 term.该提案利用哈特曼实验室的专业知识、资源和 我们为研究CYP 2 E1而生产的试剂，以及我们合作者的专业知识。的总体目标 研究计划是发现CYP 2 E1的内源性功能和调节以及 线粒体和内质网中的CYP 2 E1活性。未来五年的研究和目标概述 年：哈特曼实验室在大流行开始时在MUSC开放。它探讨的关键问题包括： 细胞如何决定将多少CYP 2 E1发送到线粒体而不是内质网？ 新生多肽靶向的机制将通过遗传和药理学来确定 方法。CYP 2 E1在多种组织中的内源性功能是什么？已知CYP 2 E1 将丙酮代谢为丙酮醇和丙酮醛，并将ω和ω-1脂肪酸羟基化 位置，但这些活动尚未在整体细胞代谢的背景下进行研究， 一般只关注肝脏。我们正在研究多种组织特异性内源性功能， 组织中线粒体和内质网中CYP 2 E1表达的后果是什么？ 我们实验室的新证据表明，对于乙醇和对乙酰氨基酚，线粒体CYP 2 E1是一种负担， 对于药物诱导的细胞损伤，但对于脂肪酸，内质网CYP 2 E1驱动脂质应激。 因此，必须进一步界定特定于组织的负债。这项工作将共同解决 关于CYP 2 E1的新出现和长期存在的基本问题。

PROJECT SUMMARY Cytochrome P450 2E1 (CYP2E1) is a unique, highly conserved, and highly regulated mammalian P450 monooxygenase. CYP2E1 undergoes extensive post-transcriptional and post-translational regulation, including bimodal targeting to endoplasmic reticulum and mitochondria. There are no known loss of function mutations in the human population, and no known polymorphic variants that change the coding region of the protein, indicating an important endogenous role for CYP2E1. However, knockout of CYP2E1 in rodent models has not yielded a dramatic phenotype, and has been reported to be protective against diet-induced obesity, nonalcoholic fatty liver disease, and ethanol-induced toxicity. Therapeutic targeting of CYP2E1 has been recommended for liver repair after alcoholic liver damage and to improve the efficacy of a ketogenic diet for the treatment of epilepsy. However, there remain major gaps in our understanding of the endogenous function of CYP2E1 that must be addressed before CYP2E1 can be safely inhibited in humans, particularly in the long- term. This proposal addresses these challenges using the expertise of the Hartman lab, resources and reagents we have generated to study CYP2E1, and the expertise of our collaborators. The overall goal of the research program is to discover the endogenous function and regulation of CYP2E1 and the consequences of CYP2E1 activity in mitochondria and endoplasmic reticulum. Overview of research and goals for the next five years: The Hartman Lab opened at MUSC during the beginning of the pandemic. It pursues key questions like: How does the cell decide how much CYP2E1 to send to mitochondria vs. the endoplasmic reticulum? Mechanisms of nascent polypeptide targeting will be determined through genetic and pharmacological approaches. What are the endogenous functions of CYP2E1 in multiple tissues? CYP2E1 is known to metabolize acetone to acetol and methylglyoxal, and to hydroxylate fatty acids at the omega and omega-1 position, but these activities have not been studied in the context of overall cellular metabolism and have generally only focused on liver. We are investigating the tissue-specific endogenous functions in multiple tissues. What are the consequences of CYP2E1 expression in mitochondria and endoplasmic reticulum? Emerging evidence in our lab suggests that for ethanol and acetaminophen, mitochondrial CYP2E1 is a liability for drug-induced cellular damage, but for fatty acids, endoplasmic reticular CYP2E1 drives lipid stress. Therefore, there are organelle-specific liabilities that must be further defined. Together this work will address arising and long-standing fundamental questions about CYP2E1.