Role of microRNA-122 in hepatocarcinogenesis using conditional knock out mice

使用条件敲除小鼠研究 microRNA-122 在肝癌发生中的作用

基本信息

批准号：
7867173
负责人：
Kalpana Ghoshal
金额：
$ 38.13万
依托单位：
OHIO STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-05-17 至 2014-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7867173
关键词：
Adult Aflatoxin B1 Age Alcohols Animal Model Antineoplastic Agents Apoptosis Aromatic Amines Aromatic Polycyclic Hydrocarbons Biological Process Cancer Detection Cancer Etiology Cancer Patient Carcinogens Carcinoma Case Study Cell physiology Cells Cessation of life Chronic Cirrhosis Complex Data Death Rate Development Diabetes Mellitus Diagnostic Neoplasm Staging Diet Diethylnitrosamine Disease Disease Progression Down-Regulation Ectopic Expression Endothelial Cells Environmental Pollutants Epigenetic Process Etiology Excision Exposure to Fatty Liver Fibrosis Figs - dietary Functional disorder Future Genes Genetic Goals Grant Growth Growth Factor Heavy Drinking Hemochromatosis Hepatic Hepatitis Hepatitis B Hepatitis B Virus Hepatitis C virus Hepatocarcinogenesis Hepatocyte Human IGF2 gene In Vitro Incidence Infection Infiltration Inflammation Inflammatory Injury Institutes Interleukin-6 Knock-out Knockout Mice Life Liposomes Liver Liver Dysfunction Liver diseases Liver neoplasms Magnetic Resonance Imaging Malignant Neoplasms Malignant neoplasm of liver MicroRNAs Mission Modeling Molecular Monitor Mus Neoplasm Metastasis Nitrosamines Nodule Nude Mice Obesity Oncogenic Operative Surgical Procedures Oral Contraceptives Patients Physiology Play Poison Predisposition Prevention Primary carcinoma of the liver cells Process Property Publishing RNA Radiation Recurrence Risk Factors Rodent Role Staging Testing Therapeutic Therapeutic Agents Tissues Treatment Efficacy Treatment Protocols Tumor Suppressor Proteins Tumor stage United States Vinyl Chloride Virus Work abstracting adenoma alpha-Fetoproteins base cancer cell cancer therapy chemical carcinogen chemotherapeutic agent chemotherapy choline deficient diet cytokine epithelial to mesenchymal transition feeding in vivo inhibitor/antagonist liver function liver transplantation loss of function men migration mimetics mortality mouse model nanoparticle non-alcoholic fatty liver novel outcome forecast preclinical study public health relevance response tumor tumor growth tumorigenic

项目摘要

DESCRIPTION (provided by applicant): The objective of this new R01 application is to advance our understanding of the role of miR-122 in hepatocarcinogenesis. HCC is the fifth most common cancer and the third common cause of cancer related death. The incidence of HCC is on the rise in USA, with little hope for cure or treatment through chemotherapy, radiation or other traditional cancer treatments. Its major risk factors are infection with hepatitis B and C viruses, and exposure to toxic chemicals, including alcohol, all of which cause chronic liver injury and inflammation. Using an animal model for hepatocarcinogenesis we were the first to demonstrate down regulation of miR-122, the most abundant liver-specific microRNA (~70% of the total miRNA) in the liver, during the initiation and progression of HCC and also in human primary HCCs. Suppression of miR-122 is a signature of HCCs with poor prognosis and metastasis. Studies with HCC cells in culture have shown that miR-122 functions as a tumor suppressor in vitro and in nude mice. To understand the biological functions of miR-122, especially in hepatocarcinogenesis, we have generated conditional knockout mice (supported by an R21 grant to the PI). These mice express 100 fold less miR-122 when crossed to AlbCre mice and spontaneously develop hepatitis in the liver with age, which is facilitated after feeding choline-deficient diet that promotes hepatocarcinogenesis. More importantly, miR-122 deleted (KO) mice are more susceptible to HCCs when exposed to diethylnitrosamine, a potent liver carcinogen. Based on these observations we hypothesize that miR-122 plays a critical role in maintaining liver function, and loss of miR-122 predisposes to liver disease including cancer. The specific aims of the proposal are: Aim 1. Investigate the role of miR-122 in a mouse model of nonalcoholic fatty liver disease (NAFLD) related HCC induced by feeding choline-deficient diet. 1a) The susceptibility of miR-122 / (KO) and miR-122fl/fl (control) mice to CDAA diet will be examined by comparing liver damage (apoptosis, steatosis or fatty liver, inflammation, fibrosis) and liver tumors (formation of adenomas and carcinomas) between these mice, and 1b) the involvement of miR-122 targets will be assessed. Aim 2. Investigate the role of miR-122 in diethylnitrosamine (DEN)-induced hepatocarcinogenesis. Pathological/molecular changes of mice injected with DEN will be monitored as described in Aim 1. Aim 3. Examine the therapeutic potential of miR-122 alone or in combination with chemotherapeutic agents to inhibit tumor growth in vivo in the DEN model. Mice will be injected weekly for 4 weeks with miR-122 mimetics loaded in galactosylated nanoparticles (to specifically target it to HCC cells) at early stages of tumor development (visualized by MRI) and the regression in the tumor growth will be compared to those in mice treated with the scrambled RNA nanoparticles. This study will elucidate the function of the most abundant liver-specific microRNA in maintaining normal liver physiology and also its therapeutic efficacy against hepatocellular carcinomas in an animal model. PUBLIC HEALTH RELEVANCE: Layman's Abstract The urgent need for new treatments for hepatocellular cancer (HCC) is warranted because incidence of HCC is increasing considerably in the United States, their dismal prognosis and the poor response of this cancer to treatment regimen currently available. This project focuses on the role of the loss of function of miR-122 in the development and progression of hepatocellular carcinoma (HCC), one of the most common human malignancies. Our previous studies have shown downregulation of miR-122 in HCCs of both human and rodent origin. Now we intend to show that the loss of function of this liver specific miR plays a causal role in the initiation and progression of HCC. If this is the case, these studies will open the way to the development of novel miR-122 therapy for this incurable disease. Thus, establishing role of miR-122 mimetic in an animal model (in preclinical trial) would be a major milestone in the treatment of this deadly disease in the near future.

描述（由申请人提供）：这种新的R01应用的目的是促进我们对miR-122在肝癌发生中的作用的理解。 HCC是第五大常见的癌症，也是癌症相关死亡的第三个常见原因。 HCC的发病率在美国的增长，通过化学疗法，放射或其他传统癌症治疗对治疗或治疗几乎没有希望。它的主要危险因素是丙型肝炎和C病毒感染，以及暴露于毒性化学物质（包括酒精）中，所有这些都会导致慢性肝损伤和炎症。使用动物模型进行肝癌发生，我们是第一个证明MiR-122调节的人，MiR-122是肝脏中最丰富的肝脏特异性microRNA（约为总miRNA的〜70％），在HCC的启动和进展过程中，在人类的启动和进展过程中。主要HCC。 miR-122的抑制是预后和转移不良的HCC的签名。在培养中对HCC细胞的研究表明，在体外和裸鼠中，miR-122充当肿瘤抑制剂。为了了解miR-122的生物学功能，尤其是在肝癌发生中，我们已经产生了条件敲除小鼠（由PI的R21授予支持）。这些小鼠在越过木板小鼠时表达100倍的miR-122，并随着年龄的增长而自发发展肝炎，这在喂食胆碱缺乏饮食后促进了促进肝癌的饮食。更重要的是，当暴露于有效的肝癌二乙基硝基胺时，miR-122删除的（KO）小鼠更容易受到HCC的影响。基于这些观察结果，我们假设miR-122在维持肝功能方面起着至关重要的作用，而miR-122的丧失易患肝病，包括癌症。该提案的具体目的是：AIM 1。研究miR-122在通过喂养胆碱缺陷饮食诱导的HCC相关的非酒精性脂肪肝疾病（NAFLD）小鼠模型中的作用。 1A）将检查miR-122 /（KO）和miR-122fl / fl（对照）小鼠对CDAA饮食的敏感性，通过比较肝脏损伤（凋亡，脂肪变性，脂肪变性或脂肪肝，炎症，纤维化）和肝肿瘤（形成肝脏肝脏）（形成）这些小鼠之间的腺瘤和癌）和1B）将评估miR-122靶标的参与。 AIM 2。研究miR-122在二乙基硝基胺（DEN）诱导的肝癌发生中的作用。如AIM 1所述，将监测注射DEN的小鼠的病理/分子变化。AIM 3。检查单独或与化学治疗剂结合使用miR-122的治疗潜力，以抑制DEN模型中体内肿瘤生长。在肿瘤发育的早期阶段（MRI可视化）将每周注射小鼠，每周注射4周的miR-122微米（以特异性靶向HCC细胞），将与肿瘤生长的回归进行比较。用炒RNA纳米颗粒处理的小鼠。这项研究将阐明最丰富的肝脏特异性microRNA在维持正常肝生理学以及其对动物模型中肝细胞癌的治疗功效方面的功能。公共卫生相关性：外行的摘要迫切需要对肝癌（HCC）进行新的治疗方法，因为在美国，HCC的发生率大大增加，它们的惨淡预后以及该癌症对当前可用治疗方案的不良反应。该项目的重点是miR-122在肝细胞癌（HCC）发展和进展中的功能丧失，这是最常见的人类恶性肿瘤之一。我们先前的研究表明，在人和啮齿动物起源的HCC中，miR-122的下调。现在，我们打算表明，该肝脏特异性miR的功能丧失在HCC的启动和进展中起因果作用。如果是这种情况，这些研究将为这种无法治愈的疾病开发新的miR-122疗法开辟道路。因此，在不久的将来，建立miR-122模拟物在动物模型中的作用（在临床前试验中）将是治疗这种致命疾病的主要里程碑。