Role of Uhrf1 in Liver Development, Regeneration and Carciogenesis

Uhrf1 在肝脏发育、再生和癌变中的作用

• 批准号: 8050096
• 负责人: Kirsten C Sadler Edepli
• 金额: $ 36.67万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-02-10 至 2014-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8050096
• 关键词: Address; Adult; Apoptosis; Biochemical; Biochemistry; Biological; Cancer Biology; Cancer cell line; Carcinogens; Cell Culture Techniques; Cell Cycle; Cell Proliferation; Cell division; Cells; Chronic; Clinical; Cultured Cells; Cyclin A; DNA Methylation Regulation; DNA Methyltransferase; DNA Modification Methylases; DNA-Binding Proteins; Data; Development; Disease; Embryo; Embryonic Development; Event; Fingers; Fishes; G1/S Transition; Gene Dosage; Gene Targeting; Genes; Genetic; Genetic Programming; Genetic Screening; Goals; Growth; Health; Hepatic; Hepatocarcinogenesis; Hepatocyte; Human; Human Biology; Human Genetics; In Vitro; Injection of therapeutic agent; Injury; Link; Liver; Liver Regeneration; Liver diseases; Liver neoplasms; Malignant Neoplasms; Malignant neoplasm of liver; Mammalian Cell; Mammals; Mediating; Modeling; Natural regeneration; Oncogenes; Oncogenic; Partial Hepatectomy; Pathologic; Pathway interactions; Patient Care; Patients; Phenotype; Phosphorylation; Physiological; Play; Predisposition; Primary carcinoma of the liver cells; Process; Property; Protein Binding; Publishing; Regulation; Role; Sampling; Suggestion; System; TOP2A gene; Techniques; Tertiary Protein Structure; Testing; Topoisomerase; Transcript; Transcription Coactivator; Transgenic Organisms; Ubiquitin; Up-Regulation; Work; Zebrafish; cancer genomics; cell transformation; chemotherapeutic agent; cohort; cost effective; cyclin A2; fatty acid-binding proteins; human CDK2 protein; human HDAC1 protein; in vivo; interest; knock-down; liver cell proliferation; loss of function mutation; mimetics; mutant; novel; promoter; response; tissue culture; tumor; tumorigenesis; ubiquitin-protein ligase; zebrafish development

DESCRIPTION (provided by applicant): Liver development in embryos and regeneration in adults are characterized by regulated hepatocyte proliferation. This contrasts with the unregulated hepatocyte proliferation that accompanies many chronic hepatic diseases and hepatocellular carcinoma (HCC). While there is suggestion that common genetic pathways regulate both physiologic and pathologic hepatocyte proliferation, only a few studies in mammals illustrate this hypothesis. By forward genetic screening in zebrafish embryos combined with functional studies on liver regeneration in adults and expression analysis of human HCC samples, we have found that the ubiquitin-like, containing PHD and RING finger domains-1 (uhrf1) gene is essential for physiologic hepatocyte proliferation. We also believe that alterations in UHRF1 expression and/or regulation also contribute to deregulated proliferation in cancer. The work in this proposal will use a combination of zebrafish development and genetics, human cancer genomic analysis and mammalian tissue culture cell cycle studies and biochemistry to address 3 aspects of UHRF1 function in relation to hepatocyte proliferation. In Specific Aim 1, we will examine the mechanism by which UHRF1 functions in regulating hepatocyte proliferation in zebrafish embryos. In Specific Aim 2, we will elucidate how UHRF1 is regulated through phosphorylation by cyclin dependent kinase 2. The functional relevance of this phosphorylation will be assessed on cultured cells using biochemical and cell biological techniques. In Specific Aim 3, we will determine the role of UHRF1 in hepatocarcinogenesis. By analysis of human HCC samples, we will evaluate the possibility that amplification of the UHRF1 locus contributes to its upregulation in cancer. Secondly, we will perform genetic and oncogenic studies in zebrafish and determine if UHRF1 is necessary and sufficient for hepatic tumor formation. In summary, this proposal will link together mechanisms that control hepatocyte proliferation in the embryo and during liver regeneration with those that control hepatocarcinogenesis. This proposal has direct relevance to the field of liver disease. Because the burden of liver disease remains enormous, the identification of genes that play critical roles in hepatocyte proliferation and in HCC progression are of significant scientific and clinical importance. The goal is to identify novel mechanisms of hepatocyte proliferation that can aid in the development of chemotherapeutic agents for use in management of patients with chronic liver diseases including HCC. PUBLIC HEALTH RELEVANCE: We are interested in how the liver develops, how it restores itself after injury and how liver cancer occurs. We believe that all three processes are linked and that understanding of normal liver growth will help in caring for patients with liver cancer. We believe that we have discovered a gene called UHRF1 that is involved in all the three processes and will study how it plays a role in each situation. PHS 398/2590 (Rev. 09/04, Reissued 4/2006) Page Continuation Format Page

描述（由申请人提供）：胚胎中的肝脏发育和成人的再生的特征是受调节的肝细胞增殖。这与许多慢性肝病和肝细胞癌 (HCC) 伴随的不受控制的肝细胞增殖形成鲜明对比。虽然有人认为共同的遗传途径调节生理和病理性肝细胞增殖，但只有少数哺乳动物研究证实了这一假设。通过对斑马鱼胚胎的正向遗传筛选，结合成人肝再生的功能研究和人类肝癌样本的表达分析，我们发现含有PHD和RING指结构域1（uhrf1）的类泛素基因对于生理肝细胞至关重要增殖。我们还认为，UHRF1 表达和/或调节的改变也有助于癌症增殖的失调。该提案中的工作将结合斑马鱼发育和遗传学、人类癌症基因组分析和哺乳动物组织培养细胞周期研究和生物化学来解决 UHRF1 功能与肝细胞增殖相关的 3 个方面。在具体目标 1 中，我们将研究 UHRF1 在调节斑马鱼胚胎中肝细胞增殖中发挥作用的机制。在具体目标 2 中，我们将阐明 UHRF1 如何通过细胞周期蛋白依赖性激酶 2 的磷酸化进行调节。将使用生化和细胞生物学技术在培养细胞上评估这种磷酸化的功能相关性。在具体目标 3 中，我们将确定 UHRF1 在肝癌发生中的作用。通过分析人类 HCC 样本，我们将评估 UHRF1 位点扩增导致其在癌症中上调的可能性。其次，我们将在斑马鱼中进行遗传和致癌研究，并确定UHRF1对于肝肿瘤的形成是否是必要和充分的。总之，该提案将控制胚胎中和肝再生过程中肝细胞增殖的机制与控制肝癌发生的机制联系起来。该提案与肝病领域直接相关。由于肝脏疾病的负担仍然巨大，因此鉴定在肝细胞增殖和 HCC 进展中发挥关键作用的基因具有重要的科学和临床意义。目标是确定肝细胞增殖的新机制，有助于开发化疗药物，用于治疗包括肝细胞癌在内的慢性肝病患者。公共健康相关性：我们对肝脏如何发育、受伤后如何自我恢复以及肝癌如何发生感兴趣。我们相信这三个过程是相互关联的，了解正常的肝脏生长将有助于护理肝癌患者。我们相信我们已经发现了一个名为 UHRF1 的基因，它参与了所有这三个过程，并将研究它如何在每种情况下发挥作用。 PHS 398/2590（修订版 09/04，重新发布 4/2006） 页面延续格式页面