FABP4 Released from Steatotic Hepatocytes in Alcoholic Liver Disease Enhances Hepatic Tumor Progression

酒精性肝病中脂肪肝细胞释放的 FABP4 促进肝肿瘤进展

• 批准号: 10380190
• 负责人: IAIN HUGH MCKILLOP
• 金额: $ 22.5万
• 依托单位: CAROLINAS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-25 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10380190
• 关键词: Acetaldehyde; Adipocytes; Adipose tissue; Affect; Alcohol consumption; Alcohol dehydrogenase; Alcohol dependence; Alcoholic Hepatitis; Alcoholic Liver Diseases; Alcohols; Animals; Antibodies; Binding; Biological Models; CYP2E1 gene; Cell Fractionation; Cell Proliferation; Cell physiology; Cells; Cellular Metabolic Process; Cellular biology; Chronic; Cirrhosis; Clinical; Consumption; Cytochromes; Data; Data Reporting; Deacetylation; Development; Diagnostic Neoplasm Staging; Diethylnitrosamine; Disease; Disease Progression; Disease model; Endocrine; Endothelial Cells; Environment; Ethanol; Ethanol Metabolism; Event; Excision; FABP1 gene; FABP4 gene; Fatty acid glycerol esters; Fibrosis; Future; Genetic Risk; Genetic Transcription; Healthcare; Hepatic; Hepatic Tissue; Hepatocyte; Hepatology; Homeostasis; Human; In Vitro; Individual; Informed Consent; Institutional Review Boards; Knowledge; Label; Letters; Lipids; Liver; Liver diseases; Liver neoplasms; MAP Kinase Gene; MAPK8 gene; Malignant Epithelial Cell; Malignant Neoplasms; Malignant neoplasm of liver; Mediating; Membrane; Metabolism; Modeling; Molecular; Molecular Biology; Molecular Chaperones; Molecular and Cellular Biology; Movement; Mus; Neoplasm Metastasis; Nonesterified Fatty Acids; Nuclear; Operative Surgical Procedures; Outcome; Oxidative Stress; Pathologic; Pathology; Pathway interactions; Patients; Pharmacology; Primary carcinoma of the liver cells; Process; Protein Isoforms; Proteins; Proteomics; Protocols documentation; Recurrence; Regulation; Reporting; Rodent; Rodent Model; Role; SIRT1 gene; Sampling; Series; Serum; Severities; Signal Pathway; Signal Transduction; Signaling Molecule; System; Tissues; Transportation; Tumor Expansion; United States; Western Blotting; alcohol use disorder; base; cell growth; cell motility; clinically relevant; exposed human population; fatty acid-binding proteins; feeding; fomepizole; hepatoma cell; in vitro Model; in vivo; inhibitor/antagonist; innovation; intrahepatic; mRNA Expression; macrophage; microvesicles; migration; mouse model; neoplastic cell; novel; paracrine; protein expression; receptor; response; therapy design; transcription factor; tumor; tumor progression

SUMMARY Alcohol use disorders (AUDs) are significant clinical and financial healthcare burdens in the United States and globally. Within the spectrum of pathologies affected by AUDs, the central role of the liver in alcohol metabolism makes it particularly susceptible to damage. Alcoholic liver disease (ALD) arises from sustained, heavy alcohol ingestion and is characterized by a series of worsening liver pathologies (fat accumulation (hepatosteatosis), alcoholic hepatitis, fibrosis-cirrhosis). In ALD, hepatic cytochrome P450 2E1 is induced to metabolize ethanol leading to elevated intrahepatic acetaldehyde and oxidative stress, factors that increase the risk of genetic damage and development of hepatocellular carcinoma (HCC). Intracellular lipid movement and storage are closely regulated processes required to maintain liver and systemic homeostasis. In healthy individuals fatty acid binding proteins (FABPs) are expressed in a tissue- specific manner and function as critical chaperones during lipid sequestration and movement. In the healthy liver, FABP1 is the predominant FABP expressed in hepatocytes. Recent studies report FABP4 (an isoform usually expressed in adipocytes and macrophages) is synthesized and released by adipocytes to act as a paracrine- endocrine signaling molecule in specific disease states, including cancers arising in close proximity to adipose tissue. Studies by our group report FABP4 mRNA and protein expression is dramatically upregulated in hepatocytes isolated from alcohol-fed rodents, and following alcohol metabolism by CYP2E1-expressing HCC cells. These findings of increased FABP4 expression in ALD model systems are also evidenced in tissue and serum from ALD/ALD-HCC patients. Functionally, we report exogenous rhFABP4 stimulates ERK-MAPK and JNK signaling leading to HCC proliferation and migration in vitro. Collectively, these data have led us to hypothesize that “alcohol metabolism in ALD-induced steatotic hepatocytes leads to the induction of FABP4 synthesis and release which in turn stimulates HCC cell growth and migration”. Two Specific Aims are proposed; Aim 1 will determine the mechanism[s] by which hepatic alcohol metabolism induces FABP4 expression, and define the signaling mechanism[s] by which FABP4 regulates hepatoma cellfunction. To achieve this, we will combine knowledge derived from our Preliminary Data using established in vitro models with innovative proteomic approaches (Cell Signaling Phospho-Antibody Array) to accurately determine the signaling networks regulated by FABP4. Aim 2 will demonstrate the significance of FABP4 signaling in alcohol-dependent HCC expansion and progression in vivo. These studies will utilize a novel hepatocyte-specific FABP4-/- mouse (HS-FABP4-/-) and an orthotopic model of tumor progression. In parallel, we will expand our ongoing analyses of tissue and serum from ALD and ALD-HCC patients for FABP4 expression to determine the clinical relevance of altered FABP4 expression during ALD and HCC progression.

总结 在美国，酒精使用障碍（AUD）是重要的临床和经济医疗保健负担 和全球范围内。在受AUD影响的病理范围内，肝脏在酒精中的中心作用 新陈代谢使它特别容易受到损害。酒精性肝病（ALD）是由持续的， 大量酒精摄入并以一系列恶化的肝脏病理（脂肪积累）为特征 （脂肪肝）、酒精性肝炎、纤维化-肝硬化）。在ALD中，肝细胞色素P450 2 E1被诱导， 代谢乙醇导致肝内乙醛和氧化应激升高，这些因素增加了 遗传损伤和肝细胞癌（HCC）发展的风险。 细胞内脂质的运动和储存是维持肝脏和肝脏功能所需的密切调节过程。 系统内稳态在健康个体中，脂肪酸结合蛋白（FABPs）在组织中表达， 在脂质螯合和运动过程中作为重要的分子伴侣的特定方式和功能。在健康的肝脏中， FABP 1是肝细胞中表达的主要FABP。最近的研究报告FABP 4（通常是一种同种型， 在脂肪细胞和巨噬细胞中表达）由脂肪细胞合成和释放，作为旁分泌， 内分泌信号分子在特定疾病状态中的作用，包括与脂肪组织密切相关的癌症 组织.我们小组的研究报告，FABP 4 mRNA和蛋白表达显著上调， 从喂食酒精的啮齿动物中分离的肝细胞，并通过表达CYP 2 E1的HCC进行酒精代谢 细胞在ALD模型系统中FABP 4表达增加的这些发现也在组织中得到证实， 来自ALD/ALD-HCC患者的血清。在功能上，我们报告外源性rhFABP 4刺激ERK-MAPK， JNK信号转导导致体外HCC增殖和迁移。总的来说，这些数据让我们 假设“酒精代谢在ALD诱导的脂肪变性肝细胞中导致FABP 4的诱导 合成和释放，其反过来刺激HCC细胞生长和迁移”。 提出了两个具体目标;目标1将确定肝醇 代谢诱导FABP 4表达，并定义FABP 4调节的信号传导机制 肝癌细胞功能为了实现这一目标，我们将使用以下方法将从初步数据中获得的联合收割机知识结合起来： 用创新的蛋白质组学方法（细胞信号磷酸化抗体阵列）建立体外模型， 准确确定FABP 4规范的信令网络。目标2将说明以下方面的重要性： FABP 4信号在体内酒精依赖性HCC扩展和进展中的作用这些研究将利用一本小说 肝细胞特异性FABP 4-/-小鼠（HS-FABP 4-/-）和肿瘤进展的原位模型。与此同时， 我们将扩大我们正在进行的对ALD和ALD-HCC患者的组织和血清的FABP 4分析， FABP 4表达的改变以确定ALD和HCC进展期间FABP 4表达改变的临床相关性。