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

基本信息

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
项目类别：
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

项目摘要

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）：关于肝脏中药物代谢酶和转运蛋白的发育调控（称为“药物加工基因” [DPGS]）知之甚少，将新生儿和儿童置于不良药物反应（ADRS）的风险更高的风险。使用RNA-seq，我们已经表明，药物代谢是无细菌（GF）小鼠整个肝脏转录组中最差异的途径，这表明肠道微生物组和肝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二级胆汁酸（BAS），该胆汁酸可以激活肝脏中的两个最关键的异种生物感应核受体，即怀孕X受体（PXR）和组成型雄激素受体（CAR）。在开发过程中，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至少可能部分有助于肝脏中DPG的发育调控。尚未进行系统的研究来表征发育过程中肠道微生物组对所有DPG的调节，并且关于如何通过抗生素或益生菌靶向肠道微生物组如何靶向肝脏中DPG的个体发育。因此，目标的目标研究是利用多学科方法，包括GF和遗传学工程小鼠，BA代谢组学，下一代测序和人类粪便样本，以揭示肠道微生物群在调节PXR和CAR信号以及随后的DPGS in liver in Liver中的pxR和CAR信号中的作用。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调节肝脏中DPG的个体发育，通过改变肠道中的次级BAS，以修改肝脏中的PXR和/或CAR信号传导。我们将在2个目标中检验我们的假设：AIM 1A将使用RNA-Seq量化GF肝脏和6个发育年龄的GF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来量化肠道细菌如何调节PXR/CAR DNA与某些DPG的结合，并将DPG与肠道微生物组（元基因组学）和BA概况（UPLC-MS/MS）的成本相关。 AIM 1B将在各种年龄段的各种PXR和CAR-knockout和人性化转基因中引入次级BAS，以测试我们的假设，即次级BAS恢复某些DPG的正常发育。 AIM 2将使用来自各种发育年龄的人类粪便细菌定殖的GF小鼠，以确定抗生素的作用和嗜酸乳杆菌在重新编写人类微生物组的个体中，以及随后在肝发育中宿主DPG的变化。 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.