Developmental regulation of HBV biosynthesis by FoxA and DNA methylation

FoxA 和 DNA 甲基化对 HBV 生物合成的发育调控

基本信息

批准号：
9275362
负责人：
Alan McLachlan
金额：
$ 39.98万
依托单位：
UNIVERSITY OF ILLINOIS AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-05-18 至 2021-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9275362
关键词：
Acute Adult Alleles Anabolism Animal Model Antiviral Therapy Capsid Cessation of life Chromatin Structure Chronic Chronic Hepatitis B Circular DNA Clinical DNA DNA Methylation DNA sequencing Development Doxycycline Embryo Epigenetic Process Family Gene Expression Gene Silencing Genes Genetic Genetic Transcription Genome Genomic DNA Health Hepatic Hepatitis B Hepatocyte Histones Human Hypermethylation Individual Lead Life Cycle Stages Liver Methylation Modality Molecular Molecular Weight Nature Nuclear Nucleotides Phenotype Physiology Play Proteins RNA Regulation Resolution Reverse Transcription Role Signal Transduction System Transcription Factor 3 Transgenes Transgenic Mice Viral Viral Genome Viral Load result Virus Diseases Virus Inactivation bisulfite cellular targeting chronic liver disease design effective therapy hepatocyte nuclear factor in vivo liver development liver function methylation pattern mouse model new therapeutic target novel novel strategies pathogen postnatal prevent targeted treatment therapeutic target transcription factor viral DNA

项目摘要

Hepatitis B virus (HBV) infection is a worldwide health problem. It is estimated that there are 200 to 500 million HBV chronic carriers in the world for whom, to date, there is no reliable treatment. HBV causes both acute and chronic liver disease and is responsible for an estimated one million deaths annually. Currently available therapies reduce viral loads but fail to resolve chronic HBV infections. Therefore, effective treatments for chronic HBV infection are urgently required. The major obstacle to the resolution of chronic HBV infections is the eradication or inactivation of nuclear HBV covalently closed circular (CCC) DNA which is the template for viral transcription. To this end, we have developed HNF1α-null HBV transgenic mice and liver-specific FoxA/HNF3-deficient HBV transgenic mice. HNF1α-null HBV transgenic mice synthesize nuclear HBV CCC DNA. Liver-specific FoxA/HNF3-deficient HBV transgenic mice are viable but lack detectable HBV transcription and replication (i.e. they are effectively “cured”) suggesting FoxA/HNF3 not only plays an important developmental role in the hepatocyte-specific transcription factor network governing the hepatic phenotype but is also essential for viral biosynthesis. The observation that FoxA/HNF3-deficient HBV transgenic mice display hyper-methylation of HBV genomic DNA suggests that FoxA/HNF3 is epigenetically governing HBV transcription by modulating viral chromatin structure in vivo and hence determines the level of viral biosynthesis. Defining the precise temporal requirements for FoxA/HNF3 expression associated with HBV transcription and replication will indicate the liver developmental stages when viral biosynthesis is susceptible to inhibition by FoxA/HNF3 deficiency. This will be achieved by modulating FoxA/HNF3 expression in the FoxA/HNF3-deficient HBV transgenic mouse model using the TET/OFF system. Using this system, the developmental control of HBV transcription, viral biosynthesis and HBV DNA methylation by FoxA/HNF3 expression will be established and correlated with the epigenetic histone marks and chromatin structure associated with the HBV genome. Finally, similar studies will be performed using the HNF1α-null FoxA/HNF3-deficient HBV transgenic mouse model of chronic viral infection so the developmental control of HBV transcription, viral biosynthesis, HBV DNA methylation, epigenetic histone marks and chromatin structure by FoxA/HNF3 expression associated with the HBV genome can be compared between the HBV transgene DNA and the nuclear HBV CCC DNA. Defining the nature of the changes and the molecular signals responsible for the loss of HBV biosynthesis due to FoxA/HNF3 deficiency may lead to the identification of cellular therapeutic targets that are amenable to the development of novel modalities to resolve rather than simply treat chronic HBV infection.

丙型肝炎病毒（HBV）感染是全球健康问题。据估计，世界上有200至5亿HBV慢性承运人，迄今为止，没有可靠的治疗方法。 HBV会引起急性和慢性肝病，并导致估计每年100万人死亡。当前可用的疗法减少病毒载荷，但无法解决慢性HBV感染。因此，迫切需要针对慢性HBV感染的有效治疗方法。慢性HBV感染的分辨率的主要障碍是核HBV共价闭合圆形（CCC）DNA的脱毛或失活，这是病毒转录的模板。为此，我们开发了HNF1α-NULL HBV转基因小鼠和肝特异性FOXA/HNF3缺乏的HBV转基因小鼠。 HNF1α-NULL HBV转基因小鼠合成核HBV CCC DNA。肝脏特异性FOXA/HNF3缺乏HBV转基因小鼠是可行的，但缺乏可检测到的HBV转录和复制（即，它们实际上是“治愈”的）表明FOXA/HNF3不仅在Hepatocyte因子网络中扮演了重要的转录因子网络在Go ni of Hepatic Perigation型中，而且还对抗肝素的现象是IS Hepation Isns Issn Is Insns Insns Is Insns Insn。 FOXA/HNF3缺乏HBV转基因小鼠表现出HBV基因组DNA的过度甲基化的观察表明，FOXA/HNF3是通过体内调节病毒染色质结构的表观遗传来控制HBV转录的，因此可以决定病毒生物合成的水平。定义与HBV转录和复制相关的FOXA/HNF3表达的精确临时要求，将表明当病毒生物合成易受FOXA/HNF3缺乏症的抑制时，肝发育阶段。这将通过使用TET/OFF系统调制FOXA/HNF3缺乏HBV转基因小鼠模型中的FOXA/HNF3表达来实现这一点。使用该系统，将建立FOXA/HNF3表达的HBV转录，病毒生物合成和HBV DNA甲基化的发展控制，并与与HBV基因组相关的表观遗传组蛋白标记和染色质结构相关。最后，将使用慢性病毒感染的HNF1α-无FOXA/HNF3缺乏HBV转基因小鼠模型进行类似的研究，从而对HBV转录，病毒生物合成，HBV DNA甲基化，表观遗传蛋白标记和HNF3表达相关的HNF CANDEN CANTIN的发育控制，HBV DNA甲基化，HNF3表达与HNF3表达相关联。核HBV CCC DNA。定义因FOXA/HNF3缺乏症引起的HBV生物合成丧失的变化的性质和分子信号可能导致鉴定细胞治疗靶标，这些靶标与开发新的方式可以解决而不是简单地治疗慢性HBV感染。