GENE EXPRESSION MODULATORS TO CONTROL DRUG METABOLISM

描述(改编自调查人员摘要)：本报告的目的 一项提议是开发基因特异性活动的潜力 药物动物模型中合成寡核苷酸的研究 新陈代谢。核酸酶抗性寡核苷酸的作用机制包括 与反基因或反义核酸的序列特异性相互作用 分子或转录调节蛋白之间的相互作用 模拟导致基因调控的基因组顺式元件 表情。研究人员打算检验这样一个假设，即最优 寡核苷酸结构可以被确定，它调节着 细胞色素P450在体内的异构体。此外，该基因表达调控 将为这些基因的表达调控提供洞察力 它们对内源性底物和肝外表达的依赖。在……里面 建议进行体内研究，因为ODN进入并在体内分布 细胞不等同于在培养细胞中进行的研究和 在完整的动物身上观察到的。这些研究的具体目的是：1) 确定反义、反基因、 核酶与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) 在体内调节剂中鉴定I期药物代谢的广谱 酶包括大鼠细胞色素P1A1、细胞色素P450 B1、细胞色素P2 B2、细胞色素P2 11、细胞色素P2 E1、细胞色素P3A2和细胞色素P3 A2 CYP4A1和4)在上下文相关的情况下评估这些基因表达调节器 性激素、昼夜节律、异物暴露所产生的表达 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构成基因的表达，2) 体内药效通过体外分析酶的活性和 蛋白质水平直接将目标mRNA与观察到的表型联系在一起，3) 毒性可以与疗效一起评估，4)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)效力、疗效和毒性的直接比较 由线性硫代磷酸脱氧核糖核酸制成。未来的研究将涉及更多 详细研究细胞色素P450在卵巢癌中的表达 体内氧自由基敏感基因的调控。

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.