PXR: Regulation and Pharmacogenomics

PXR：监管和药物基因组学

• 批准号: 6879186
• 负责人: ERIN G SCHUETZ
• 金额: $ 29.41万
• 依托单位: ST. JUDE CHILDREN'S RESEARCH HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-03-01 至 2008-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6879186
• 关键词: bioluminescence; cytochrome P450; genetic promoter element; genetic regulation; genetically modified animals; human tissue; laboratory mouse; ligands; liver cells; luciferin monooxygenase; messenger RNA; pharmacogenetics; phenotype; polymerase chain reaction; pregnane compound; protein sequence; protein structure function; receptor expression; rifamycins; steroid hormone receptor; transcription factor

DESCRIPTION (provided by applicant): Adverse drug interactions secondary to induction of drug detoxification genes by ligand activated PXR is an important clinical problem leading to decreased drug efficacy. Despite our ability to use rapid in vitro screens to identify compounds that activate PXR, there is still no way to predict the extent of drug interaction in individual patients. We believe that human variation in PXR expression contributes to the interindividual differences in induction of CYP3A. Moreover, the growing list of drug detoxification genes induced by PXR further justifies the need to determine what is responsible for variable PXR expression. Our prior studies have revealed that sequence variation (SNPs, single nucleotide polymorphisms) in the PXR coding region is not significantly contributing to human variation in PXR expression or human variation in induction of the CYP3A target gene. Our current research is designed to further elucidate the cellular and molecular mechanisms regulating PXR expression. Our prior studies have revealed that there are multiple isoforms of PXR that differ in their amino terminus and in the ligand binding domain. Our current research in Aim 1 is designed to discover the extent to which PXR with different amino termini and different ligand binding domains are expressed in human tissues and whether these PXR splice variants have altered function. We hypothesize that these variant PXRs fine tune PXR activation creating an additional layer of regulation, and ultimately influence the CYP3A inductive phenotype. Towards the goal of understanding factors regulating PXR expression Aims 2 and 4 will determine the role of liver enriched transcriptional factors in regulation of human PXR transcription and PXR expression in human liver. In order to visualize PXR transcriptional activity in mouse tissues, in Aim 3 we will develop a transgenic mouse with the PXR-promoter-driving a luciferase reporter. Bioluminescent imaging through the skin of the mouse will monitor in real-time PXR transcription under the most informative biological conditions-i.e., in a living intact animal. This unique and powerful whole body imaging approach will allow us to visualize PXR transcription in multiple tissues under different physiological and drug treated environments. In total this proposal describes a comprehensive set of experiments to determine how PXR is regulated.

描述（由申请方提供）：继发于配体激活PXR诱导药物解毒基因的药物不良相互作用是导致药物疗效降低的重要临床问题。尽管我们有能力使用快速体外筛选来鉴定激活PXR的化合物，但仍然没有办法预测个体患者中药物相互作用的程度。我们认为，人类PXR表达的差异有助于诱导CYP3A的个体间差异。此外，PXR诱导的药物解毒基因越来越多，这进一步证明了需要确定什么是可变PXR表达的原因。我们之前的研究已经揭示，PXR编码区的序列变异（SNP，单核苷酸多态性）对PXR表达的人类变异或CYP3A靶基因诱导的人类变异没有显著影响。我们目前的研究旨在进一步阐明调节PXR表达的细胞和分子机制。我们先前的研究表明，有多种异构体的PXR，不同的氨基末端和配体结合域。我们目前在目标1中的研究旨在发现具有不同氨基末端和不同配体结合结构域的PXR在人体组织中表达的程度，以及这些PXR剪接变体是否具有改变的功能。我们假设这些变异的PXR微调PXR激活，产生额外的调节层，并最终影响CYP3A诱导表型。为了理解调节PXR表达的因子，目的2和4将确定肝脏富集的转录因子在人肝脏中人PXR转录和PXR表达的调节中的作用。为了可视化小鼠组织中的PXR转录活性，在目标3中，我们将开发具有PXR启动子驱动荧光素酶报告基因的转基因小鼠。通过小鼠皮肤的生物发光成像将在信息量最大的生物条件下实时监测PXR转录，即，在一个完整的动物身上。这种独特而强大的全身成像方法将使我们能够在不同的生理和药物治疗环境下可视化多个组织中的PXR转录。总之，该提案描述了一套全面的实验，以确定PXR是如何调节的。