Mechanisms that promote hepatocellular carcinoma due to chronic ethanol exposure

长期接触乙醇促进肝细胞癌的机制

• 批准号: 10666121
• 负责人: PAMELA L. TUMA
• 金额: $ 18.43万
• 依托单位: CATHOLIC UNIVERSITY OF AMERICA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-05-05 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10666121
• 关键词: 8q24; Affect; Aflatoxins; Alcohol consumption; Alcohols; Animal Model; Automobile Driving; Benign; Biochemical; Cell Line; Cells; Cellular biology; Cholangiocarcinoma; Chromosomes; Chronic; Cirrhosis; Combined Modality Therapy; Coupled; Cultured Cells; Drug resistance; Epithelium; Etiology; Excision; Exposure to; Fluorescent in Situ Hybridization; Genomic Instability; Goals; Hepatitis; Hepatitis B Infection; Hepatitis C; Hepatitis C virus; Hepatocyte; Human; In Vitro; Label; Malignant Epithelial Cell; Malignant Neoplasms; Malignant neoplasm of liver; Mesenchymal; Monitor; Morphology; Neoplasm Metastasis; Paraffin Embedding; Pathology; Patients; Pattern; Phenotype; Primary Neoplasm; Primary carcinoma of the liver cells; Prognosis; Renal Cell Carcinoma; Repression; Resources; Services; Site; Stains; Survival Rate; Testing; Tissue Sample; Tissues; alcohol exposure; c-myc Genes; chronic alcohol ingestion; chronic liver disease; clinically relevant; cohort; diagnostic biomarker; drug sensitivity; epithelial to mesenchymal transition; experimental study; gene repression; human tissue; imaging facilities; knock-down; lymph nodes; medical schools; member; novel therapeutics; overexpression; patient response; patient subsets; prevent; prognostic; protein expression; stemness; trait; transcriptome; translational impact; translational study; treatment response; treatment strategy

SUMMARY Hepatocellular carcinoma (HCC) is the fifth most common cancer world-wide with over 600,000 new cases per year and a dismal five-year survival rate at less than 9%. Most HCC patients have chronic liver disease resulting mainly from HCV/HBV infection, chronic alcohol consumption or aflatoxin exposure. Although most patients that develop HCC have cirrhosis or hepatitis, the mechanisms promoting HCC differ between pathologies such that not all cases should be treated the same. This proposal is aimed at understanding the mechanisms that promote HCC due to chronic ethanol exposure, a form of HCC that is on the rise as HCV and HBV infections are being better prevented and treated. In particular, this proposal is focused on understanding chromosome 8q24 amplification that is observed more frequently in alcohol-induced HCC. Because c-Myc resides on this amplified region, we propose that at later stages of HCC, chromosome 8q24 is amplified leading to high c-Myc levels. The high Myc promotes Miz1 transcriptional repression and loss of the epithelial phenotype and acquisition of mesenchymal cell traits. Concomitantly, the high c-Myc activates Zeb1 expression driving cells through the epithelial to mesenchymal transition (EMT) further repressing the epithelial phenotype. As cells metastasize and colonize at secondary sites, epithelial traits are partially reacquired as cells progress through the mesenchymal to epithelial transition (MET). Our hypothesis is that the temporal expression patterns of c-Myc, Zeb1 and Miz1 and epithelial vs. mesenchymal traits are unique in patients with alcohol-induced HCC and can predict prognosis, drug resistance and potential treatment strategies. In Aim 1 we ask two major questions: 1) does 8q24 amplification drive EMT in alcohol-associated HCC? and 2) do metastatic lesions reacquire a more epithelial phenotype? To answer these questions, we will examine protein expression patterns using immunohistochemical (IHC) staining of paraffin-embedded human HCC resections (to answer question 1) and affected lymph nodes and metastatic lesions (to answer question 2). Staining will be correlated to cancer stage, clinicopathologic variables, alcohol consumption and patient responses to treatment. We will also confirm 8q24 amplification using FISH analysis of c-Myc expression. To directly determine the temporal relationship between c-Myc, Miz1, Zeb1 and the epithelial vs. mesenchymal phenotypes, we will perform targeted transcriptome analysis and morphological analysis of human benign and HCC tissue. We will also perform overexpression and knockdown studies to directly establish the temporal relationship between c-Myc, Miz1, Zeb1 and epithelial vs. mesenchymal phenotypes in selected cell lines and primary human normal and HCC cells in culture. Finally, due to the tremendous need to develop new strategies for effective combination therapies to treat HCC, we will examine drug sensitivity in cells to clinically relevant compounds. We have garnered the support of several others to provide their expertise to the project. We have continued access to the Imaging Facility located at nearby Johns Hopkins Medical School and are members of the Hopkins GI Center allowing us access to many services and resources. The expansive expertise of our collaborators and the access to human tissue samples and high- end resources coupled with our considerable expertise in polarized hepatic cell biology situate us perfectly to perform these important mechanistic and translational studies.

概括 肝细胞癌 (HCC) 是全球第五大常见癌症，每年新增病例超过 600,000 例 一年的生存率和五年生存率低于 9%。大多数肝癌患者患有慢性肝病 主要来自HCV/HBV感染、长期饮酒或黄曲霉毒素暴露。虽然大多数患者认为 发展为 HCC 并伴有肝硬化或肝炎，不同病理情况下促进 HCC 的机制有所不同，例如 并非所有情况都应得到同样的对待。该提案旨在了解促进的机制 由于长期接触乙醇而导致的 HCC，随着 HCV 和 HBV 感染的流行，这种 HCC 的发病率呈上升趋势。 更好地预防和治疗。特别是，该提案的重点是了解染色体 8q24 放大现象在酒精诱发的 HCC 中更为常见。因为 c-Myc 驻留在这个扩增的 区域，我们认为在 HCC 的后期阶段，染色体 8q24 被扩增，导致高 c-Myc 水平。这 高 Myc 促进 Miz1 转录抑制和上皮表型丧失以及获得 间充质细胞特征。同时，高 c-Myc 通过以下方式激活 Zeb1 表达驱动细胞 上皮间质转化（EMT）进一步抑制上皮表型。随着细胞的转移和 在次生位点定植，随着细胞通过间充质细胞的进展，上皮特征被部分重新获得 到上皮转化（MET）。我们的假设是 c-Myc、Zeb1 和 Miz1 和上皮与间质特征在酒精诱发的 HCC 患者中是独特的，并且可以 预测预后、耐药性和潜在的治疗策略。在目标 1 中，我们提出两个主要问题： 1) 8q24 扩增是否会驱动酒精相关性 HCC 中的 EMT？ 2) 转移性病灶是否会重新获得更多的 上皮表型？为了回答这些问题，我们将使用以下方法检查蛋白质表达模式 石蜡包埋的人 HCC 切除物的免疫组织化学 (IHC) 染色（回答问题 1）和 受影响的淋巴结和转移病灶（回答问题 2）。染色与癌症分期相关， 临床病理变量、饮酒量和患者对治疗的反应。我们还将确认 8q24 使用 c-Myc 表达的 FISH 分析进行扩增。直接确定之间的时间关系 c-Myc、Miz1、Zeb1 和上皮与间质表型，我们将进行靶向转录组 人类良性和肝癌组织的分析和形态学分析。我们还将进行过度表达和 敲低研究直接建立 c-Myc、Miz1、Zeb1 和上皮细胞与上皮细胞之间的时间关系。 选定细胞系以及培养的原代人类正常细胞和肝癌细胞中的间充质表型。最后， 由于迫切需要开发治疗 HCC 的有效联合疗法的新策略，我们将 检查细胞对临床相关化合物的药物敏感性。我们已经获得了其他几个人的支持 为该项目提供他们的专业知识。我们可以继续使用位于附近的成像设施 约翰霍普金斯医学院，并且是霍普金斯胃肠病中心的成员，使我们能够获得许多服务 和资源。我们的合作者拥有广泛的专业知识以及获取人体组织样本和高 最终资源加上我们在极化肝细胞生物学方面的丰富专业知识使我们能够完美地 进行这些重要的机制和转化研究。