Developmental pathways, environmental agents, and epigenetics in liver disease

描述（由申请人提供）：肝细胞癌（HCC）是全球第五大常见癌症，估计每年造成600，000例死亡。酒精滥用和丙型肝炎病毒（HCV）感染引起炎症和慢性肝病，包括肝硬化，并协同增加HCC的风险。肝硬化是一种明确的癌前病变，但其在表观基因组水平上的分子基础尚未得到研究。许多国家的酒精消费量正在上升，每年造成约180万人死亡。全世界约有1.7亿人感染HCV，其中美国有400万人，使其成为美国最常见的血液传播疾病。大多数HCC（>85%）的病原体可能比任何其他癌症更容易理解，所有这些都是由于环境因素或人类行为造成的（例如酒精，HCV），但我们对这些药物如何在>30年的时间内发挥其致癌作用并具有显著的临床变异性仍有许多不了解。没有明确的证据表明酒精或HCV直接介导遗传损伤。然而，越来越多的证据表明，肝病期间的早期和进行性表观遗传变化以及饮食和炎症在破坏表观基因组中的作用。根据我们小组和其他人的研究，调节发育的基因似乎是癌症（包括肝癌）表观遗传缺陷的优先靶点之一。从未在正常或患病肝脏中对发育途径的表观遗传调控进行过全面分析，限制了我们对人类肝脏疾病病因学表观遗传变化的完整程度和时间以及这些变化如何用作预后标志物以改善治疗的了解。我们推测，慢性酒精暴露和/或HCV感染引起的发育途径的表观遗传变化在肝脏疾病，HCV诱导的细胞损伤和HCC发展的发病机制中起着关键作用。我们提出了四个目标来解决这个假设。在目标1中，我们将通过将大型肝组织库与专注于发育调控基因的定制平铺阵列耦合，对来自酒精和/或HCV引起的肝硬化或HCC患者的人类肝组织进行表观遗传学分析。在目标2中，我们将使用细胞培养系统对未处理和酒精处理/HCV感染的原代肝细胞进行表观遗传学分析。在目标3中，我们将用灵敏和定量的测定来证实我们的阵列数据，并研究乙醇/HCV调节模型发育基因的表观遗传控制的机制，以及发育基因的异常表达如何调节细胞生长。最后，在目标4中，我们将研究发育基因的表观遗传标记/表达如何与肝病进展期间的临床和病理变化相关。这些研究的结果有望大大提高我们对HCC表观遗传病因学的理解，为治疗和诊断/预后提供新的靶点，并产生关于无处不在的娱乐性药物和广泛传播的感染性病原体如何影响表观基因组的新信息。

DESCRIPTION (provided by applicant): Hepatocellular carcinoma (HCC) is the 5th most common cancer worldwide, causing an estimated 600,000 deaths/yr. Alcohol abuse and hepatitis C virus (HCV) infection cause inflammation and chronic liver diseases, including cirrhosis, and synergistically increase risk for HCC. Cirrhosis is a well-defined precancerous lesion, yet its molecular underpinnings at the level of the epigenome have not been examined. Alcohol consumption is rising in many countries and results in ~1.8 million deaths/yr. Approximately 170 million people worldwide are infected with HCV, 4 million in the United States, making it the most common blood-borne disease in the U.S. The etiologic agents responsible for the majority of HCCs (>85%) are perhaps better understood than for any other cancer, and all are due to environmental agents or human behaviors (e.g. alcohol, HCV), yet there is much that we do not understand about how these agents exert their carcinogenic effects over >30 years with significant clinical variability. There is no definitive evidence for alcohol or HCV directly mediating genetic damage. There is mounting evidence, however, for early and progressive epigenetic changes during liver disease and for a role of diet and inflammation in disrupting the epigenome. Based on studies from our group and others, genes regulating development appear to be one of the preferential targets of epigenetic defects in cancers, including HCC. A comprehensive analysis of the epigenetic regulation of developmental pathways has never been performed in normal or diseased liver, limiting our knowledge of the full extent and timing of epigenetic changes in the etiology of human liver disease and how these changes may be used as prognostic markers to improve treatment. We hypothesize that epigenetic changes in developmental pathways caused by chronic alcohol exposure and/or HCV infection play a key role in the pathogenesis of liver disease, HCV-induced cellular damage, and HCC development. We propose four aims to address this hypothesis. In aim 1 we will epigenetically profile human liver tissues from patients with cirrhosis or HCC caused by alcohol and/or HCV by coupling a large liver tissue bank with a custom tiling array focused on developmental regulatory genes. In aim 2 we will epigenetically profile untreated and alcohol treated/HCV infected primary hepatocytes using a cell culture system. In aim 3 we will confirm our array data with sensitive and quantitative assays and examine the mechanisms by which ethanol/HCV modulate epigenetic control of model developmental genes and how aberrant expression of developmental genes regulates cell growth. Finally, in aim 4 we will examine how epigenetic marks/expression of developmental genes relates to clinical and pathological changes during liver disease progression. Results from these studies are expected to greatly enhance our understanding of the epigenetic etiology of HCC, provide new targets for therapy and diagnosis/prognosis, and yield novel information on how a ubiquitous recreational drug and a widespread infectious agent impact the epigenome.