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（根据调查员的摘要进行了改编）：目的 建议是利用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。 抗核酸酶抗性ODN的机械作用包括 序列特异性与核酸作为抗固体或反义的相互作用 分子或转录调节蛋白的方式 模仿基因组元素导致基因调节 表达。 研究人员打算检验最佳的假设 可以鉴定寡核苷酸结构调节 cytochrome P450 isoforms in vivo. 此外，基因表达调制 将提供有关这些基因表达的调节的见解和 他们对内源性底物和肝外表达的依赖。 在 提出了体内研究，因为ODN进入和分布 在培养细胞中进行的研究之间，细胞不是等效的， 在完整动物中观察到。 这些研究的具体目的是：1） 确定最佳的寡核苷酸结构，用于反义，抗基因， 核酶和体内基因表达的转录调节，2） 评估这些基因表达调节剂的作用机理，3） 在体内调节剂中识别一系列I期药物代谢 包括大鼠CYP1A1，CYP2B1，CYP2B2，CYP2C11，CYP2E1，CYP2E1，CYP3A2和 CYP4A1和4）在上下文中评估这些基因表达调节剂 性激素，昼夜节律，异种生物暴露产生的表达 和肝外器官。 该模型系统的优点是1） constitutive gene expression is monitored in the absence of disease, 2) the 体内疗效通过体外分析酶活性和 蛋白水平将靶mRNA与观察到的表型直接连接起来，3） 毒性可以与功效同时评估，4）方法是 成本效益，5）这种方法避免了细胞中的差异 培养和6）可以直接比较效力，功效和毒性 用线性磷酸座ODN制成。 未来的研究将涉及更多 细胞色素p450表达在中的作用的详细研究 调节体内自由基氧敏感基因。