Cell Signaling, PXR Phosphorylation & Drug Disposition

细胞信号转导、PXR 磷酸化

• 批准号: 7095870
• 负责人: Jeffrey L Staudinger
• 金额: $ 25.75万
• 依托单位: UNIVERSITY OF KANSAS LAWRENCE
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-08-01 至 2010-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7095870
• 关键词: cytochrome P450; cytokine; drug metabolism; drug receptors; gene induction /repression; genetic promoter element; inflammation; laboratory mouse; laboratory rat; liver cells; molecular site; nuclear receptors; nuclear transport; pharmacogenetics; pharmacokinetics; phosphorylation; protein kinase A; protein kinase C; protein localization; protein protein interaction; protein structure function; receptor expression; site directed mutagenesis; tissue /cell culture; transcription factor

DESCRIPTION (PROVIDED BY APPLICANT): Drug disposition is a highly regulated process in mammals. For example, the expression of genes encoding drug-transporting and drug-metabolizing proteins is induced by their substrates in liver and intestine. Recent research has shed new light on the molecular mechanism of this phenomenon. The pregnane X receptor (PXR) is a ligand-activated transcription factor that is expressed in liver and intestine. PXR coordinately regulates the drug-inducible expression of drug-transporting and drug-metabolizing proteins in liver and intestine. PXR binds to xenobiotic-response elements (XREs) in the promoters of its target genes as a heterodimer with the 9-cis-retinoic acid receptor (RXR). One extremely important PXR-target gene is the cytochrome P450 3A4 (CYP3A4) gene. CYP3A4 is primarily expressed in liver and intestine where it catalyzes the oxidative metabolism of nearly 60% of all clinically prescribed drugs. The hepatic expression of CYP3A4 and other PXR-target genes is rapidly suppressed in response to inflammatory cytokines, though the molecular basis of this repression is currently unknown. Repression of PXR-target gene expression occurs in people that undergo invasive surgery in response to inflammation, thereby producing altered pharmacokinetic properties of drugs in these patients. PXR is activated by a plethora of structurally diverse molecules including drugs, steroids, bile acids, and xenobiotics. Therefore, repression of PXR-target gene expression can produce potentially life-threatening drug-drug interactions. Thus, it is important to understand the molecular basis of the repression of PXR-target genes in order to predict and prevent the occurrence of drug interactions in post-operative patients. An alteration in the phosphorylation status of transcriptional regulatory proteins is a likely mechanism that mediates the repression of PXR-target gene expression following inflammation. The goals of these investigations are to (1) identify the sites of PXR phosphorylation following activation of specific cell signaling pathways, and (2) determine the effect of increased PXR phosphorylation on it's ability to (a) transactivate, (b) bind DNA, (c) translocate from the cytoplasm to the nucleus, and (d) interact with protein co-factors. Successful completion of these studies will allow us to determine the molecular basis of the repression of key drug metabolizing genes during the inflammatory response and provide increased opportunities to understand the xenobiotic response. These studies will provide additional insight into the regulation of drug disposition and drug-drug interaction.

性状（申请人提供）：药物处置在哺乳动物中是一个高度调节的过程。例如，编码药物转运蛋白和药物代谢蛋白的基因的表达由它们在肝和肠中的底物诱导。最近的研究对这一现象的分子机制有了新的认识。胆甾烷X受体（PXR）是一种在肝脏和肠道中表达的配体激活的转录因子。PXR协同调节肝和肠中药物转运和药物代谢蛋白的药物诱导表达。PXR与其靶基因启动子中的异源反应元件（XRE）结合，作为具有9-顺式-视黄酸受体（RXR）的异源二聚体。一个非常重要的PXR靶基因是细胞色素P450 3A 4（CYP 3A 4）基因。CYP 3A 4主要在肝脏和肠道中表达，在那里它催化近60%的临床处方药物的氧化代谢。CYP 3A 4和其他PXR靶基因的肝脏表达在对炎症细胞因子的反应中被迅速抑制，尽管这种抑制的分子基础目前尚不清楚。PXR靶基因表达的抑制发生在那些因炎症而接受侵入性手术的人身上，从而改变了这些患者的药物药代动力学特性。PXR被大量结构多样的分子激活，包括药物、类固醇、胆汁酸和外源性物质。因此，PXR靶基因表达的抑制可能产生潜在的危及生命的药物相互作用。因此，重要的是要了解PXR靶基因的抑制的分子基础，以预测和预防药物相互作用在术后患者的发生。转录调节蛋白磷酸化状态的改变是介导炎症后PXR靶基因表达抑制的可能机制。这些研究的目的是（1）确定特定细胞信号传导途径激活后PXR磷酸化的位点，和（2）确定增加的PXR磷酸化对其（a）反式激活，（B）结合DNA，（c）从细胞质易位到细胞核，和（d）与蛋白质辅因子相互作用的能力的影响。这些研究的成功完成将使我们能够确定在炎症反应过程中抑制关键药物代谢基因的分子基础，并提供更多的机会来了解异生素反应。这些研究将为药物处置和药物相互作用的调节提供更多的见解。