权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Developmental Regulation of Drug Processing Genes

药物加工基因的发育调控

基本信息

批准号：
9060713
负责人：
Yue Cui
金额：
$ 58.41万
依托单位：
UNIVERSITY OF WASHINGTON
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-05-03 至 2017-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9060713
关键词：
Developmental Regulation Drug Processing Genes

项目摘要

DESCRIPTION (provided by applicant): Children are particularly susceptible to adverse effects of drugs and environmental chemicals due to their immature ability to process xenobiotics. Supported by the parent R01 grant, the investigators are actively studying developmental regulation of drug processing genes (DPGs) in mice. Their long-term goal is to understand species differences and similarities in xenobiotic metabolism and hepatotoxicity, a huge challenge in drug development and risk assessment of human exposure to environmental chemicals. The objective of this proposal is to recruit new collaborators with expertise in stem-cell/cancer research, bioengineering, and bioinformatics to establish a transdisciplinary team and a virtual consortium to develop a novel 3D culture model of hepatocyte differentiation and maturation from stem cells to bridge the huge gap in translating scientific findings from animals to humans. The current bottleneck in hepatocytes differentiation from stem cells in vitro is that the differentiated hepatocytes are largely immature hepatocytes with very low expression of DPGs, which are required to evaluate xenobiotic metabolism and hepatotoxicity. 3D culture provides a microenvironment essential for cell differentiation and hepatocyte function. Growth hormone (GH) plays a key role in postnatal liver development. Oxygen availability is essential in differentiation of stem cells and metabolic function of hepatocytes. Hypoxia-inducible factor 1a (HIF-1?) and NF-?B are master regulators of hypoxic response. Hepatocyte nuclear factor 4? (HNF4?) is a master regulator of hepatocyte differentiation and liver function. ?-catenin plays a key role in not only liver morphogenesis, but also hepatic basal expression and induction of DPGs. There are extensive interactions among HIF-1?, NF-?B, HNF4?, and ?-catenin in gene regulation; however, how they interact and regulate hepatocyte differentiation and maturation during liver development is unknown. The central hypothesis is that 3D microenvironment, oxygen availability, and GH are essential in hepatocyte maturation via affecting interactions among HIF-1?, NF-?B, HNF4?, Stat5, ?-catenin, and the ?-catenin effector LEF-1. This central hypothesis will be tested in 3 specific aims. Aim 1 will use ChIP- sequencing to determine the interactions among HIF-1?, NF-?B, HNF4?, Stat5, ?-catenin, and LEF-1 in regulating gene expression during hepatocyte differentiation and maturation in mouse liver and a 3D culture model. Aim 2 will determine the physical interaction between HNF4? and LEF-1/TCF4 and effects of ?-catenin modulators on the differentiation and maturation of hepatocytes from mouse stem cells. Aim 3 will determine effects of a 3D nanofibrous scaffold, oxygen tension, and GH on the maturation of hepatocytes differentiated from mouse stem cells. This novel 3D culture model of hepatocyte differentiation and maturation from stem cells will be an invaluable tool to elucidate species differences and similarities between humans and mice in: 1) developmental regulation of DPGs; 2) effects of environmental chemicals and therapeutic drugs on hepatocyte differentiation and maturation during liver development; and 3) metabolism and hepatotoxicity of xenobiotics.

描述（由申请人提供）：由于儿童处理外源性物质的能力不成熟，他们特别容易受到药物和环境化学品的不良影响。在父母R 01基金的支持下，研究人员正在积极研究小鼠药物加工基因（DPG）的发育调控。他们的长期目标是了解异生物质代谢和肝毒性的物种差异和相似性，这是药物开发和人类暴露于环境化学品风险评估的巨大挑战。该提案的目的是招募具有干细胞/癌症研究，生物工程和生物信息学专业知识的新合作者，以建立一个跨学科团队和一个虚拟联盟，开发一种新的干细胞肝细胞分化和成熟的3D培养模型，以弥合将科学发现从动物转化为人类的巨大差距。目前在体外从干细胞分化肝细胞的瓶颈是分化的肝细胞主要是具有非常低的DPG表达的未成熟肝细胞，这是评估异生物质代谢和肝毒性所需的。3D培养提供了细胞分化和肝细胞功能所必需的微环境。生长激素（GH）在出生后肝脏发育中起着关键作用。氧供应对于干细胞的分化和肝细胞的代谢功能是必不可少的。缺氧诱导因子1a（HIF-1？）而NF-？B是低氧反应的主要调节因子。肝细胞核因子4？(HNF4?)是肝细胞分化和肝功能的主要调节剂。?-连环蛋白不仅在肝脏形态发生中起关键作用，而且在肝脏基础表达和DPG诱导中起关键作用。HIF-1？之间存在广泛的相互作用，NF-？B，HNF 4？，然后呢？连环蛋白在基因调控中起重要作用;然而，它们在肝脏发育过程中如何相互作用并调节肝细胞分化和成熟尚不清楚。核心假设是，三维微环境，氧的可用性，和生长激素是必不可少的肝细胞成熟，通过影响HIF-1？，NF-？B，HNF 4？，Stat5，？-连环蛋白，还有？连环蛋白效应子LEF-1。这一中心假设将在3个具体目标中进行检验。目的1将利用ChIP-测序技术确定HIF-1？，NF-？B，HNF 4？，Stat5，？-连环蛋白和LEF-1在小鼠肝脏和3D培养模型中调节肝细胞分化和成熟过程中的基因表达。目的2将确定HNF 4？和LEF-1/TCF 4的影响。连环蛋白调节剂对小鼠干细胞肝细胞分化和成熟的影响。目的3将确定3D纳米纤维支架、氧张力和GH对从小鼠干细胞分化的肝细胞成熟的影响。这种从干细胞分化和成熟肝细胞的新型3D培养模型将是阐明人类和小鼠在以下方面的物种差异和相似性的宝贵工具：1）DPG的发育调节; 2）环境化学品和治疗药物对肝脏发育期间肝细胞分化和成熟的影响;以及3）外源性物质的代谢和肝毒性。