MicroRNA involvement in the regulation of biliary epithelium

MicroRNA参与胆管上皮的调节

• 批准号: 8470161
• 负责人: Florin Selaru
• 金额: $ 15.87万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-01 至 2016-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8470161
• 关键词: Adhesions; Algorithms; Apoptosis; Behavior; Biliary; Cell Adhesion; Cell Cycle; Cell Cycle Progression; Cell Cycle Regulation; Cell Line; Cell physiology; Cells; Characteristics; Cholangiocarcinoma; Data; Disease; Epithelium; Functional RNA; Funding; Gene Expression Regulation; Genes; Goals; Growth; Homeostasis; Human; In Vitro; Incidence; Inflammatory; Knock-out; Laboratories; Link; Liver; MADH4 gene; Malignant Neoplasms; Measures; Messenger RNA; MicroRNAs; Mitotic Cell Cycle; Modeling; Molecular Biology; Mus; Neoplasm Metastasis; Neoplasms; Neoplastic Cell Transformation; PTEN gene; Pathway interactions; Patients; Phenotype; Physiology; Play; Principal Investigator; Proteins; Regulation; Role; Stress; Testing; Training; United States; Xenograft Model; Xenograft procedure; biliary tract; cell behavior; cell growth; cholangiocyte; hepatobiliary cancer; in vivo; in vivo Model; inhibitor/antagonist; mRNA Differential Displays; meetings; neoplastic; new therapeutic target; primary sclerosing cholangitis; public health relevance; research study; response

DESCRIPTION (provided by applicant): MicroRNAs (miRs) are a new class of small non-coding RNAs that may regulate up to 30% of human genes. miRs are involved in a myriad of cellular processes: proliferation, differentiation, apoptosis and responses to stress, and were shown to be involved in normal cholangiocyte physiology as well as in cholangiocarcinomas (CCA). CCAs are cancers arising from the biliary tract. They are the 2nd most common primary hepatobiliary cancers in the United States, and their incidence is increasing globally. The survival of patients with CCA is dismal, usually measured in months. Our preliminary studies in cholangiocytes identified one hundred and thirty two miR species with differential expression in cholangiocytes vs. neoplastic cholangiocytes. We hypothesize that among these miRs, there are several miR species that have an etiologic role in neoplasia formation and homeostasis. Furthermore, we hypothesize that these miRs have important role in the normal cholangiocyte homeostasis, regulating cell cycle progression, proliferation and/or apoptosis. In the current application, we propose a step- wise approach to identify miRs with phenotypic impact, followed by careful characterization of their function. Specifically, we propose to: 1. Identify 4 miRs that have a phenotypic impact on cholangiocytes in vitro (Aim 1). We will select miR species that have similar phenotypic impact on human (Aim 1a) and murine (Aim 1b) cell lines. The reason for this restriction is that we plan on using the SMAD4/PTEN murine model of cholangiocarcinoma for our in vivo studies. Next, we propose to 2. Further validate the in vitro phenotypic action of the candidate miRs through in vivo studies (Aim 2). By applying a step-wise algorithm, we plan on verifying the miR action in a xenograft model (Aim 2a) and then to further study the miRs with phenotypic impact in a liver-specific SMAD4/PTEN-deleted murine model of CCA. At the conclusion of Aims 1 and 2, we will have identified 4 miR species with phenotypic impact (cell cycle, proliferation, apoptosis) on cholangiocytes in vitro and in vivo. These miR species are very likely to be involved in normal cholangiocyte cellular processes as well as altered as cholangiocytes become neoplastic. Finally, we propose to 3. Identify mechanism of action for miR species identified in Aim 1 and confirmed in Aim 2 (Aim 3). We will identify mRNA and protein targets for these miRs as well as explore the pathways connecting these miRs to their mRNA targets. I believe that through funding this application and execution of the proposed experiments, several goals will be met: 1. my training in molecular biology will be perfected with the final goal of contributing to my independence as principal investigator; 2. the current understanding of the normal physiology of cholangiocytes as well as the neoplastic transformation will be advanced. Ultimately, I am hopeful that through the findings of this project, we will make significant progress towards understanding how and why inflammatory conditions of the biliary tree, such as Primary Sclerosing Cholangitis, progress to neoplasia.

描述（由申请人提供）：microRNA（mirs）是一类新的非编码RNA，可调节多达30％的人类基因。 miR参与了无数的细胞过程：增殖，分化，凋亡和对压力的反应，并被证明参与了正常的胆管细胞生理学以及胆管癌（CCA）。 CCA是由胆道引起的癌症。它们是美国第二大最常见的原发性肝胆碱癌，它们的发病率在全球范围内增加。 CCA患者的生存期很沮丧，通常在几个月内测量。 我们在胆管细胞中的初步研究鉴定了一百三十种MiR物种在胆管细胞中具有差异表达与肿瘤胆管细胞。我们假设在这些miR中，有几种miR物种在肿瘤形成和稳态中具有病因。此外，我们假设这些miR在正常的胆管细胞稳态中具有重要作用，从而调节细胞周期的进展，增殖和/或凋亡。在当前的应用中，我们提出了一种分步方法，以识别具有表型影响的miR，然后仔细地表征其功能。具体而言，我们提出：1。识别4个对体外胆管细胞具有表型影响的miR（AIM 1）。我们将选择对人（AIM 1A）和鼠（AIM 1B）细胞系具有相似表型影响的miR物种。这种限制的原因是我们计划将胆管癌的Smad4/Pten鼠模型用于体内研究。接下来，我们提议2。通过体内研究进一步验证候选miR的体外表型作用（AIM 2）。通过应用逐步算法，我们计划在异种移植模型（AIM 2A）中验证MIR作用，然后进一步研究具有表型影响的MIR在肝脏特异性SMAD4/PTEN删除的CCA模型中。在目标1和2的结论中，我们将确定4种具有表型影响（细胞周期，增殖，细胞凋亡）对胆管细胞的体外和体内的miR物种。这些miR物种很可能参与正常的胆管细胞细胞过程，并且随着胆管细胞的变化而变化。最后，我们建议3。确定在AIM 1中鉴定并在AIM 2中确认的MIR物种的作用机理（AIM 3）。我们将确定这些miR的mRNA和蛋白质靶标，并探索将这些miR与它们的mRNA靶标连接的途径。 我认为，通过资助此应用程序和拟议实验的执行，将实现几个目标：1。我在分子生物学方面的培训将得到完善，这是为我作为首席研究员独立的最终目标； 2。当前对胆管细胞正常生理的理解以及肿瘤转化的理解。最终，我希望通过该项目的发现，我们将取得重大进展，以了解胆汁树的炎症状况以及为什么诸如原发性硬化性胆管炎的炎症状况以及为什么发展为肿瘤。