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

• 批准号: 8446381
• 负责人: Kalpana Ghoshal
• 金额: $ 30.23万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-17 至 2015-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8446381
• 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; Functional disorder; Future; Genes; 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; 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; Mutation; 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 inflammation; liver injury; liver transplantation; loss of function; men; migration; mimetics; mortality; mouse model; nanoparticle; non-alcoholic fatty liver; nonalcoholic steatohepatitis; novel; outcome forecast; preclinical study; public health relevance; response; tumor; tumor growth; tumorigenic

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.

描述（由申请人提供）：这项新的 R01 申请的目的是增进我们对 miR-122 在肝癌发生中作用的理解。 HCC 是第五大常见癌症，也是癌症相关死亡的第三大常见原因。在美国，肝癌的发病率正在上升，通过化疗、放疗或其他传统癌症治疗方法治愈或治疗的希望渺茫。其主要危险因素是感染乙型和丙型肝炎病毒，以及接触有毒化学物质，包括酒精，所有这些都会导致慢性肝损伤和炎症。我们利用肝癌发生的动物模型，首次证明了在 HCC 以及人类原发性 HCC 的发生和进展过程中，肝脏中最丰富的肝脏特异性 microRNA（约占总 miRNA 的 70%）miR-122 的下调。 miR-122 的抑制是预后不良和转移的 HCC 的标志。对培养中的 HCC 细胞的研究表明，miR-122 在体外和裸鼠体内发挥肿瘤抑制因子的作用。为了了解 miR-122 的生物学功能，特别是在肝癌发生中的生物学功能，我们培育了条件性基因敲除小鼠（由 PI 的 R21 资助支持）。当与 AlbCre 小鼠杂交时，这些小鼠的 miR-122 表达量减少了 100 倍，并且随着年龄的增长，肝脏中会自发出现肝炎，而在喂养缺乏胆碱的饮食后会促进肝癌的发生。更重要的是，miR-122缺失（KO）小鼠在接触二乙基亚硝胺（一种强效肝癌物质）时更容易患肝癌。基于这些观察结果，我们假设 miR-122 在维持肝功能中发挥着关键作用，而 miR-122 的缺失易导致包括癌症在内的肝病。该提案的具体目标是： 目的 1. 研究 miR-122 在由缺乏胆碱饮食诱导的非酒精性脂肪性肝病 (NAFLD) 相关 HCC 小鼠模型中的作用。 1a) 通过比较这些小鼠之间的肝损伤（细胞凋亡、脂肪变性或脂肪肝、炎症、纤维化）和肝肿瘤（腺瘤和癌的形成）来检查 miR-122/（KO）和 miR-122fl/fl（对照）小鼠对 CDAA 饮食的敏感性，1b）将评估 miR-122 靶标的参与情况。目标 2. 研究 miR-122 在二乙基亚硝胺 (DEN) 诱导的肝癌发生中的作用。将如目标 1 中所述监测注射 DEN 的小鼠的病理/分子变化。目标 3. 检查 miR-122 单独或与化疗药物组合在 DEN 模型中抑制体内肿瘤生长的治疗潜力。在肿瘤发展的早期阶段（通过 MRI 观察），小鼠将每周注射半乳糖基化纳米颗粒中装载的 miR-122 模拟物（将其特异性靶向 HCC 细胞），持续 4 周，并将肿瘤生长的消退情况与用乱序 RNA 纳米颗粒治疗的小鼠进行比较。这项研究将阐明最丰富的肝脏特异性 microRNA 在维持正常肝脏生理机能方面的功能及其在动物模型中对肝细胞癌的治疗功效。

项目成果

Lactosylated gramicidin-based lipid nanoparticles (Lac-GLN) for targeted delivery of anti-miR-155 to hepatocellular carcinoma.

• DOI: 10.1016/j.jconrel.2013.03.020
• 发表时间: 2013-06-28
• 期刊: Journal of controlled release : official journal of the Controlled Release Society
• 作者: Zhang M;Zhou X;Wang B;Yung BC;Lee LJ;Ghoshal K;Lee RJ
• 通讯作者: Lee RJ

MicroRNAs in Liver Health and Disease.

• DOI: 10.1007/s40139-012-0005-4
• 发表时间: 2013-03
• 期刊: Current pathobiology reports
• 作者: Hsu SH;Ghoshal K
• 通讯作者: Ghoshal K

Cationic lipid nanoparticles for therapeutic delivery of siRNA and miRNA to murine liver tumor.

• DOI: 10.1016/j.nano.2013.05.007
• 发表时间: 2013-11
• 期刊: NANOMEDICINE-NANOTECHNOLOGY BIOLOGY AND MEDICINE
• 影响因子: 5.4
• 作者: Hsu, Shu-hao;Yu, Bo;Wang, Xinmei;Lu, Yuanzhi;Schmidt, Carl R.;Lee, Robert J.;Lee, L. James;Jacob, Samson T.;Ghoshal, Kalpana
• 通讯作者: Ghoshal, Kalpana