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）：此目的 提议是开发基因特异性活动的潜力 涉及药物的动物模型中的合成寡核苷酸（ODN） 代谢。 核酸酶抗性 ODN 的作用机制包括 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以这样的方式 模拟基因组顺式元件，导致基因调节 表达。 研究人员打算检验最优假设 可以鉴定调节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 它们对内源性底物和肝外表达的依赖。 在 提出体内研究是因为 ODN 进入并在体内分布 在培养细胞中进行的研究之间并不等同 在完整的动物中观察到。 这些研究的具体目标是：1) 鉴定反义、反基因的最佳寡核苷酸结构， 体内基因表达的核酶和转录调控，2) 评估这些基因表达调节剂的作用机制，3) 识别体内调节剂的广谱 I 期药物代谢 酶包括大鼠 CYP1A1、CYP2B1、CYP2B2、CYP2C11、CYP2E1、CYP3A2 和 CYP4A1 和 4) 根据上下文评估这些基因表达调节剂 由性激素、昼夜节律、外源性物质暴露产生的表达 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) 体内功效通过酶活性的体外分析得到证实 蛋白质水平将目标 mRNA 与观察到的表型直接联系起来，3) 毒性可以与功效一起评估，4) 该方法是 成本效益高，5) 这种方法避免了单元格中的差异 培养和 6) 效价、功效和毒性的直接比较 由线性硫代磷酸酯 ODN 制成。 未来的研究将涉及更多 详细研究细胞色素 P450 表达在 体内自由基氧敏感基因的调节。