Developmental Regulation of Drug Metabolism by Targeting the Gut Microbiome

通过靶向肠道微生物群对药物代谢的发育调节

• 批准号: 8750942
• 负责人: Yue Cui
• 金额: $ 29.74万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-10 至 2019-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8750942
• 关键词: Adult; Age; Antibiotics; Area; Attention; Bacteria; Bile Acids; ChIP-seq; Child; Childhood; DNA Binding; Development; Drug Interactions; Drug Kinetics; Drug Regulations; Drug Targeting; Drug usage; Enzymes; Epigenetic Process; Food-Drug Interactions; Gene Expression Process; Gene Expression Profile; Genes; Genetic; Genetic Polymorphism; Genetically Engineered Mouse; Genomics; Germ-Free; Goals; Growth; Hepatic; Human; Human Microbiome; Individual; Individual Differences; Infant; Integration Host Factors; Intestines; Knock-out; Knowledge; Laboratory Animals; Lactobacillus acidophilus; Lead; Ligands; Link; Literature; Lithocholic Acid; Liver; Mediating; Messenger RNA; Metabolic syndrome; Metagenomics; Mission; Mus; Neomycin; Newborn Infant; Nuclear Receptors; Obesity; Outcome; Pathway interactions; Patients; Pharmaceutical Preparations; Probiotics; Process; Processed Genes; Proteins; Quality of life; Reaction; Receptor Signaling; Regulation; Research; Role; Sampling; Testing; Transgenic Organisms; Variant; Work; Xeno; Xenobiotic Metabolism; Xenobiotics; constitutive androstane receptor; cytochrome P450 3A; drug mechanism; drug metabolism; frontier; gut microbiota; high risk; improved; interdisciplinary approach; metabolomics; microbiome; next generation sequencing; novel; pediatric pharmacology; pregnane X receptor; programs; public health relevance; sensor; transcription factor; transcriptome sequencing

DESCRIPTION (provided by applicant): Very little is known about the developmental regulation of drug-metabolizing enzymes and transporters (together called "drug-processing genes" [DPGs]) in liver, placing newborns and children at a much higher risk of adverse drug reactions (ADRs). Using RNA-Seq, we have shown that drug metabolism is the top most differentially regulated pathway in the entire liver transcriptome of germ-free (GF) mice, suggesting that there is a novel interaction between gut microbiome and hepatic DPGs. One of the key functions of gut microbiome is to produce secondary bile acids (BAs), which can activate two most critical xenobiotic-sensing nuclear receptors in liver, namely the pregnane X receptor (PXR) and constitutive androstane receptor (CAR). During development, profound changes occur in the intestinal bacteria and the secondary BA profiles, suggesting that gut microbiome may at least in part contribute to the developmental regulation of DPGs in liver. No systematic studies have been performed to characterize the regulation of all DPGs by gut microbiome during development, and little is known regarding how targeting the gut microbiome by antibiotics or probiotics re- programs the ontogeny of DPGs in liver. Therefore the goal of this research is to utilize multidisciplinary approaches, including GF and genetically-engineered mice, BA metabolomics, Next-Generation Sequencing, and human fecal samples, to unveil the role of gut microbiota in modulating PXR and CAR signaling and the subsequent ontogenic re-programming of DPGs in liver. Our central hypothesis is: the developmental changes in the gut microbiome at least in part contribute to the regulation of the ontogeny of DPGs in liver, through altering secondary BAs in the gut to modify the PXR and/or CAR signaling in liver. We will test our hypothesis in 2 Aims: Aim 1A will use RNA-Seq to quantify mRNAs of 281 critical DPGs in livers of GF and conventional (Conv) mice at 6 developmental ages, and validate the proteins and activities of differentially regulated DPGs. We will also use ChIP-Seq to quantify how gut bacteria modulate PXR/CAR DNA binding to certain DPGs, and correlate DPG ontogeny with the ontogeny of gut microbiome (metagenomics) and BA profiles (UPLC-MS/MS). Aim 1B will introduce secondary BAs to GF mice in various PXR- and CAR-knockout and humanized transgenic) at various ages to test our hypothesis that secondary BAs restore the normal ontogeny of certain DPGs. Aim 2 will use GF mice colonized with human fecal bacteria from various developmental ages, to determine the roles of antibiotics and the probiotic L. acidophilus in re-programming the ontogeny of human microbiome and the subsequent changes in the host DPGs during liver development. The proposed work will unveil a novel link between the ontogeny of gut microbiome and the developmental changes of drug- processing capacities during development, and will lead to a paradigm shift in pediatric pharmacology, by establishing a new concept in considering ADRs in children, which are the "bug-drug" interactions, in addition to the known "drug-drug" and "food-drug" interactions.

描述（由申请人提供）：对于肝脏中药物代谢酶和转运蛋白（统称为“药物加工基因”[DPG]）的发育调节知之甚少，这使新生儿和儿童面临更高的药物不良反应（ADR）风险。使用 RNA-Seq，我们发现药物代谢是无菌 (GF) 小鼠整个肝脏转录组中最受差异调节的途径，这表明肠道微生物组和肝脏 DPG 之间存在新的相互作用。肠道微生物组的关键功能之一是产生次级胆汁酸 (BA)，它可以激活肝脏中两种最关键的异生素感应核受体，即孕烷 X 受体 (PXR) 和组成型雄甾烷受体 (CAR)。在发育过程中，肠道细菌和次级 BA 谱发生了深刻的变化，这表明肠道微生物组可能至少部分有助于肝脏中 DPG 的发育调节。尚未进行系统研究来表征发育过程中肠道微生物组对所有 DPG 的调节，并且对于抗生素或益生菌如何靶向肠道微生物组重新编程肝脏中 DPG 的个体发育知之甚少。因此本次活动的目标 该研究旨在利用多学科方法，包括 GF 和基因工程小鼠、BA 代谢组学、下一代测序和人类粪便样本，揭示肠道微生物群在调节 PXR 和 CAR 信号传导以及随后肝脏中 DPG 个体重新编程中的作用。我们的中心假设是：肠道微生物组的发育变化至少部分有助于肝脏中 DPG 个体发育的调节，通过改变肠道中的次级 BA 来修改肝脏中的 PXR 和/或 CAR 信号传导。我们将在 2 个目标中检验我们的假设：目标 1A 将使用 RNA-Seq 定量 6 个发育年龄的 GF 和传统 (Conv) 小鼠肝脏中 281 个关键 DPG 的 mRNA，并验证差异调节的 DPG 的蛋白质和活性。我们还将使用 ChIP-Seq 量化肠道细菌如何调节 PXR/CAR DNA 与某些 DPG 的结合，并将 DPG 个体发育与肠道微生物组个体发育（宏基因组学）和 BA 谱 (UPLC-MS/MS) 相关联。目标 1B 将在不同年龄的各种 PXR 和 CAR 敲除和人源化转基因小鼠中引入次级 BA，以测试我们的假设，即次级 BA 恢复某些 DPG 的正常个体发育。目标 2 将使用定植有不同发育年龄的人类粪便细菌的 GF 小鼠，以确定抗生素和益生菌嗜酸乳杆菌在重新编程人类微生物组的个体发育以及肝脏发育过程中宿主 DPG 的后续变化中的作用。拟议的工作将揭示肠道微生物组的个体发育与发育过程中药物处理能力的发育变化之间的新联系，并将通过建立考虑儿童 ADR 的新概念（除了已知的“药物与药物”和“食物与药物”相互作用之外，即“虫子与药物”相互作用），导致儿科药理学的范式转变。