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Molecular regulation of hepatic cell differentiation and maturation

肝细胞分化和成熟的分子调控

基本信息

批准号：
10456054
负责人：
Stacey S Huppert
金额：
$ 40.28万
依托单位：
CINCINNATI CHILDRENS HOSP MED CTR
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-09-23 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10456054
关键词：
Acute Liver Failure Adopted Adult Architecture Binding Binding Sites Birth CREBBP gene Cell Differentiation process Cell Maturation Cell model Cell physiology Cells ChIP-seq Childhood Chromatin Chromatin Remodeling Factor Complex Coupled DNA DNA Binding Defect Derivation procedure Development Drug toxicity Embryo Embryonic Development Endoderm Enhancers Environment Etiology Failure Gene Activation Gene Expression Gene Expression Regulation Gene Targeting Genes Genetic Transcription Genomics Goals HNF4A gene Hepatic Hepatocyte Hepatotoxicity Histone H2A Human In Vitro Knock-in Mouse Knowledge Lead Liver Liver Failure Molecular Morphology Mus Newborn Infant Organ Physiological Play Pregnancy Process Proteins Publishing Regulation Repression Research Role Site Specific qualifier value Time Tissue Engineering Zinc Fingers chromatin remodeling embryonic stem cell fetal gene repression genetic signature healing in vivo induced pluripotent stem cell insight liver function loss of function mouse model postnatal progenitor programs response to injury transcription factor

项目摘要

PROJECT SUMMARY Our application builds on the published knowledge that during embryonic development until after birth, specified hepatocytes undergo a process of differentiation where they adopt the physiological functions and morphology associated with the adult liver. Our preliminary studies deleting Zinc finger HIT-type containing 1 (Znhit1) at mid-gestation in embryonic hepatoblasts demonstrates a specific and essential role in the postnatal liver for survival, normal cellular architecture, and molecular gene signatures without changing the expression of the six key hepatic cell master regulators. Znhit1's proposed role as a core subunit of the Snf2-Related CREB-binding protein Activator Protein (SRCAP)-chromatin remodeling complex may initiate the necessary changes in chromatin architecture through insertion of the alternative histone H2A.Z to influence gene expression. The goal of this application is to determine whether Znhit1 allows or disrupts access of transcription factors to different gene targets and/or enhancers, and thereby provides a switch towards hepatic cell differentiation. We hypothesize that Znhit1, part of the SRCAP complex, regulates the hepatocyte differentiation-dependent transcriptional program. In Aim 1 we will determine whether Znhit1's impact on liver function is due to 1) an autonomous hepatoblast and/or hepatocyte defect and 2) master regulator access to gene targets and/or enhancers. For this we will use in vivo mouse models to perform temporal hepatoblast and hepatocyte deletion of Znhit1, and to determine if the hepatoblast and/or hepatocyte transcriptional program and genomic binding sites of master regulators such as Hnf4a and Foxa2 are impacted. In Aim 2 we will define what regulates the developmental switch for hepatocyte adult master regulator function. To determine whether Znhit1 is part of the switch to enable repression of the fetal and/or activation of the adult hepatocyte program, we will use AAV/DJ8-Ttr-Znhit1-GPF to force expression of Znhit1 at specific time points in mouse models and in culture iPSC-generated hepatocyte-like cells. In Aim 3, we will determine whether H2A.Z is incorporated into specific sites in a differentiation-dependent manner regulated by Znhit1. To answer this question, we will use a Znhit1-3xFlag-P2A-Zsgreen knock-in mouse to perform Znhit1 ChIP-Seq and compare to Znhit1 dependent H2A.Z bound sites. Our research will generate new insight into the molecular regulation of hepatic cell identity and differentiation.

项目总结