Cellular Signaling in Drug-induced Toxicity

药物引起的毒性中的细胞信号转导

基本信息

批准号：
8417385
负责人：
Namandje N Bumpus
金额：
$ 30.78万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-04-01 至 2018-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8417385
关键词：
Acetylcysteine Acute Liver Failure Adverse event Anabolism Anti-Retroviral Agents Antioxidants Antiviral Agents Apoptosis Apoptotic Attenuated BIM Bcl-2-binding protein Bax protein Biochemical Biological Assay Biological Models Boxing Cell Death Cessation of life Clinical Markers Coupled Data Enzymes Event Exhibits Gene Expression Profiling Genes Genetic Genetic Transcription Glutathione Goals Hepatocyte Hepatotoxicity Histology Human Immunoblotting In Vitro Infection Knockout Mice Lead Macaca Mass Spectrum Analysis Measures Mediating Metabolism Modeling Molecular Mus Nevirapine Oxidative Stress Parents Pathway interactions Patients Pharmaceutical Preparations Prevention Prevention strategy Proteins Purine Nucleotides RNA Interference Reporter Genes Reporting Resistance Ritonavir Rodent Role Sampling Signal Pathway Signal Transduction Small Interfering RNA Techniques Testing Therapeutic Toxic effect Up-Regulation Virus Diseases Work antiretroviral therapy base cytotoxicity efavirenz glucose metabolism in vitro Model in vivo inhibitor/antagonist liquid chromatography mass spectrometry liver injury mRNA Expression metabolomics new therapeutic target novel novel strategies prevent protein expression public health relevance research study

项目摘要

DESCRIPTION (provided by applicant): Efavirenz (EFV) and nevirapine (NVP) are currently prescribed antiretroviral drugs of the same class that are hepatotoxic. Recently, a major role for cellular signaling pathways in modulating toxicity downstream of EFV metabolism has been reported in vitro using primary human hepatocytes. Increased oxidative stress and the upregulation of the proapoptotic protein BimEL (Bcl-2 interacting mediator of cell death extra long) have been demonstrated to be crucial in the stimulation of EFV-induced cell death, and NVP appears to modulate hepatocyte death via the same mechanism. The goal of this proposal is to determine the manner in which EFV and NVP upregulate BimEL, while also gaining a mechanistic understanding of the events downstream of BimEL that mediate the hepatocyte death. The aims are as follows: (1) Test whether genetic deficiency of proapoptotic BimEL results in protection against EFV- and NVP-mediated cell death in vivo: BimEL null mice will be used to determine whether the absence of BimEL renders mice resistant to hepatotoxicity stimulated by these compounds; RNA interference studies in hepatocytes will be used to determine the role of effector proteins, Bax and Bak, that are downstream of BimEL in modulating EFV- and NVP-induced hepatocyte death; siRNA and reporter gene assays will be used to define the mechanism by which EFV and NVP regulate the transcription of BimEL; (2) determine if proapoptotic pathways that are activated by antiretrovirals in primary human hepatocytes are also activated in macaques receiving antiretroviral therapy; (3) probe a role for alteration of the glutathione pathway in EFV- and NVP-induced hepatotoxicity; the effects of these compounds on the glutathione pathway will be examined using a mass spectrometry-based approach that will precisely quantitate the intermediates/products of this pathway; RNA interference will be used to define the role of enzymes in the glutathione pathway in EFV- and NVP-induced hepatotoxicity. It is expected that completion of the proposed studies will lead to the identification of novel therapeutic targets for the potential prevention and treatment of EFV- and NVP-mediated hepatotoxicity.

描述（由申请人提供）：Efavirenz（EFV）和Nevirapine（NVP）目前是肝毒性毒性的同一类的抗逆转录病毒药物。最近，使用原代人肝细胞在体外据报道了细胞信号通路在调节EFV代谢下游的毒性中的主要作用。氧化应激的增加和促凋亡蛋白Bimel的上调（Bcl-2相互作用的细胞死亡介质超长）已证明对于刺激EFV诱导的细胞死亡至关重要，NVP似乎通过相同的机制调节了肝细胞死亡。该提案的目的是确定EFV和NVP上调Bimel的方式，同时还对Bimel下游的事件有一种机械理解，该事件介导了介导肝细胞死亡的情况。目的如下：（1）测试息肉性BIMEL的遗传缺乏是否会导致对体内EFV和NVP介导的细胞死亡的保护：Bimel null小鼠将使用这些化合物抑制了对抗肝毒性的小鼠是否缺乏Bimel的小鼠；肝细胞中的RNA干扰研究将用于确定Bimel在调节EFV和NVP诱导的肝细胞死亡方面的下游效应蛋白，BAX和BAK的作用； siRNA和记者基因测定将用于定义EFV和NVP调节Bimel转录的机制。（2）确定在接受抗逆转录病毒疗法的猕猴中也激活了原代人肝细胞中抗逆转录病毒的促凋亡途径；（3）探测EFV和NVP诱导的肝毒性中谷胱甘肽途径改变的作用；这些化合物对谷胱甘肽途径的影响将使用基于质谱的方法检查，该方法将精确量化该途径的中间体/产物。 RNA干扰将用于定义酶在EFV和NVP诱导的肝毒性中的作用。预计拟议的研究的完成将导致鉴定新的治疗靶标，以预防和治疗EFV和NVP介导的肝毒性。