Cellular Signaling in Drug-induced Toxicity

药物引起的毒性中的细胞信号转导

• 批准号: 9206162
• 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
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-04-01 至 2019-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9206162
• 关键词: 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; Cell Death; Cessation of life; Clinical Markers; Coupled; Cysteine; Data; Enzymes; Event; Exhibits; Gene Expression Profiling; Genes; Genetic; Genetic Transcription; Glutathione; Goals; Hepatocyte; Hepatotoxicity; Histology; Human; Immunoblotting; In Vitro; Infection; Knockout Mice; 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; Viral; Virus Diseases; Work; antiretroviral therapy; base; cytotoxicity; efavirenz; experimental study; 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

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）：依法韦仑（EFV）和奈韦拉平（NVP）是目前处方的同类抗逆转录病毒药物，具有肝毒性。最近，在体外使用原代人肝细胞报道了细胞信号通路在调节 EFV 代谢下游毒性中的主要作用。氧化应激的增加和促凋亡蛋白 BimEL（细胞死亡超长 Bcl-2 相互作用介质）的上调已被证明对于刺激 EFV 诱导的细胞死亡至关重要，并且 NVP 似乎通过相同的机制调节肝细胞死亡。该提案的目标是确定 EFV 和 NVP 上调 BimEL 的方式，同时获得对 BimEL 下游介导肝细胞死亡事件的机制理解。目的如下： (1) 测试促凋亡 BimEL 的遗传缺陷是否会导致体内免受 EFV 和 NVP 介导的细胞死亡的保护：将使用 BimEL 无效小鼠来确定 BimEL 的缺失是否使小鼠对这些化合物刺激的肝毒性具有抵抗力；肝细胞中的 RNA 干扰研究将用于确定效应蛋白 Bax 和 Bak 的作用，它们是 BimEL 下游在调节 EFV 和 NVP 诱导的肝细胞死亡中的作用； siRNA 和报告基因检测将用于定义 EFV 和 NVP 调节 BimEL 转录的机制； (2)确定在原代人肝细胞中被抗逆转录病毒药物激活的促凋亡途径在接受抗逆转录病毒治疗的猕猴中是否也被激活； (3) 探讨谷胱甘肽途径的改变在 EFV 和 NVP 诱导的肝毒性中的作用；将使用基于质谱的方法检查这些化合物对谷胱甘肽途径的影响，该方法将精确定量该途径的中间体/产物； RNA 干扰将用于确定谷胱甘肽途径中酶在 EFV 和 NVP 诱导的肝毒性中的作用。预计拟议研究的完成将导致确定新的治疗靶点，用于潜在预防和治疗 EFV 和 NVP 介导的肝毒性。