GENE EXPRESSION MODULATORS TO CONTROL DRUG METABOLISM

控制药物代谢的基因表达调节剂

• 批准号: 2865274
• 负责人: PATRICK L IVERSEN
• 金额: $ 10.68万
• 依托单位: OREGON STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-06-01 至 2001-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2865274
• 关键词: antisense nucleic acid; cytochrome P450; drug metabolism; enzyme activity; enzyme induction /repression; erythromycin; laboratory rat; methyltransferase; microsomes; midazolam; nitrophenol; northern blottings; oligonucleotides; oxygenases; polymerase chain reaction; ribozymes; western blottings

DESCRIPTION (Adapted from Investigator's Abstract): The purpose of this proposal is to exploit the potential for gene-specific activities of synthetic oligonucleotides (ODNs) in an animal model involving drug metabolism. The mechanistic actions of nuclease resistant ODNs include sequence-specific interactions with nucleic acids as antigene or antisense molecules or with transcriptional regulatory proteins in a manner that mimics the genomic cis-elements resulting in the modulation of gene expression. The investigators intend to test the hypothesis that optimal oligonucleotide structures can be identified that modulate the expression of cytochrome P450 isoforms in vivo. Further, that gene expression modulation will provide insights into the regulation of these genes' expression and their reliance on endogenous substrates and extrahepatic expression. In vivo studies are proposed because ODN entry into and distribution within the cell is not equivalent between studies conducted in cultured cells and that observed in intact animals. The specific aims of these studies are to: 1) identify optimal oligonucleotide structures for antisense, antigene, ribozyme and transcriptional regulation of gene expression in vivo, 2) evaluate the mechanism of action of these modulators of gene expression, 3) identify in in vivo modulators a broad spectrum of phase I drug metabolizing enzymes including rat CYP1A1, CYP2B1, CYP2B2, CYP2C11, CYP2E1, CYP3A2 and CYP4A1 and 4) evaluate these gene expression modulators in context dependent expression created by sex hormones, circadian rhythm, xenobiotic exposure and extrahepatic organs. The advantages of this model system are that 1) constitutive gene expression is monitored in the absence of disease, 2) the in vivo efficacy is confirmed by in vitro analysis of enzyme activities and protein levels directly link the target mRNA with observed phenotype, 3) toxicity can be evaluated concomitantly with efficacy, 4) the approach is cost effective, 5) this approach avoids discrepancies that are in cell culture and 6) direct comparison of potency, efficacy and toxicity can be made with linear phosphorothioate ODNs. Future studies will involve a more detailed investigation of the role of cytochrome P450 expression in the regulation of radical oxygen sensitive genes in vivo.

描述（改编自研究者摘要）：本研究的目的 建议是利用基因特异性活动的潜力， 在涉及药物的动物模型中合成寡核苷酸（ODN） 新陈代谢. 核酸酶抗性ODN的机制作用包括 与作为反基因或反义的核酸的序列特异性相互作用 分子或转录调节蛋白， 模拟基因组顺式元件，导致基因调控 表情 研究人员打算检验一个假设， 可以鉴定调节以下表达的寡核苷酸结构： 体内细胞色素P450亚型。 此外，基因表达调节 将提供对这些基因表达调控的见解， 它们依赖于内源性底物和肝外表达。 在 提出体内研究是因为ODN进入和分布在 在培养细胞中进行的研究之间不等同， 在完整动物中观察到。 这些研究的具体目标是：1） 鉴定反义、反基因 核酶和体内基因表达的转录调控，2） 评估这些基因表达调节剂的作用机制，3） 在体内鉴定广谱I相药物代谢调节剂 包括大鼠CYP1A1、CYP2B1、CYP2B2、CYP2C11、CYP2E1、CYP3A2和 CYP4A1和4）在背景依赖性中评价这些基因表达调节剂 性激素、昼夜节律、外源性暴露引起的表达 和肝外器官。 该模型系统的优点是：1） 在没有疾病的情况下监测组成型基因表达，2） 通过酶活性的体外分析证实体内功效， 蛋白质水平直接将靶mRNA与观察到的表型联系起来，3） 毒性可以与疗效同时评估，4）该方法 成本效益，5）这种方法避免了单元格中的差异 培养和6）直接比较效力、功效和毒性， 用线性硫代磷酸ODN制备。 未来的研究将涉及更多 详细研究细胞色素P450表达的作用， 体内自由基氧敏感基因的调节。