Does the RNA editor Apobec-1 shift microRNA targeting to a tumorigenic effect?

RNA 编辑器 Apobec-1 是否会将 microRNA 靶向转变为致瘤作用？

• 批准号: 8648159
• 负责人: Dewi Harjanto
• 金额: $ 5.15万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-01 至 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8648159
• 关键词: 3' Untranslated Regions; Adenosine; Affect; Amino Acids; Area; Automobile Driving; Binding; Bioinformatics; Biological Assay; Cancer Model; Cataloging; Catalogs; Categories; Cells; Characteristics; Chromatin; Code; Custom; Cytosine; DNA; DNA Methylation; DNA Sequence; Data; Deaminase; Development; Disease; Disease Outcome; Disease Progression; Enterocytes; Enzymes; Epigenetic Process; Exhibits; Functional disorder; Gastrointestinal tract structure; Gene Expression; Gene Expression Profile; Gene Mutation; Genes; Genetic; Genomics; Glioma; Goals; High-Throughput Nucleotide Sequencing; Human; Immunoprecipitation; Inosine; Intestinal Cancer; Intestines; Investigation; Knock-out; Life; Link; Literature; Liver; Malignant Neoplasms; Mammalian Cell; Mediating; Methods; MicroRNAs; Mining; Modeling; Modification; Molecular Biology Techniques; Mus; Oncogenic; Outcome; Phenotype; Play; RNA; RNA Editing; RNA Sequences; RNA Splicing; Regulation; Research Personnel; Role; Site; Small Intestinal Adenoma; Testing; Tissues; Transcript; Translations; Tumor Burden; Tumorigenicity; Untranslated Regions; Uracil; Work; adenoma; base; cancer cell; chromatin remodeling; crosslink; gain of function; histone modification; interest; macrophage; mouse model; neoplastic cell; next generation sequencing; novel; novel diagnostics; public health relevance; research study; tool; transcriptome sequencing; tumor; tumor progression; tumorigenesis; tumorigenic

PROJECT SUMMARY According to the current paradigm, cancer originates in dysfunction at the DNA level. As such, studies into the epigenetic regulation of cancer have historically focused on the effects of DNA methylation, histone modifications, and chromatin remodeling (the last two being outcomes of DNA mutation). However, given that mutational signatures at the DNA level are insufficient to classify many subtypes of cancer, researchers have also begun exploring the role that RNA epigenetics may play in cancer progression. One potential RNA epigenetic mechanism of interest is RNA editing, or the site-specific modification of transcribed RNA by deaminases. RNA editing is known to affect splicing, amino acid coding, and microRNAs (miRNAs) and their targets. Very recently, evidence has emerged to support a direct causative role for adenosine to inosine (A-to- I) RNA editing in cancer (Chen et al., 2013). Here, I seek to determine if cytosine to uracil (C-to-U) editing, mediated by Apobec-1, can play causative roles in cancer progression. Genetic experiments had previously suggested an involvement of Apobec-1 in disease progression in the Apcmin/+ mouse model of intestinal cancer. I am therefore proposing studies to understand the mechanistic role of Apobec-1 mediated RNA editing in the context of cancer progression, in this tumor model. My preliminary data in primary mammalian cells suggest that C-to-U editing can affect translation levels of edited transcripts by modifying miRNA target sites. I hypothesize that Apobec-1-mediated RNA editing promotes a more aggressive phenotype in tumor cells by altering miRNA targeting to affect gene expression. This work will be accomplished by exploiting next generation sequencing, bioinformatics, and molecular biology techniques. Using small intestinal tissue from Apcmin/+ mice, I will: (a) catalog C-to-U edit sites throughout the transcriptome; (b) identify potential miRNA target sites affected by Apobec-1 editing, focusing on those occurring in potentially cancer-related genes; and (c) perform functional assays to evaluate the significance of the identified targets. These experiments aim to establish a direct mechanistic link between Apobec-1-mediated RNA editing and cancer. In addition, they will also provide evidence for a novel mechanism of cancer progression that could extend to all cancers, and will yield both novel diagnostic tools and chemotherapeutic targets. !

项目概要 根据目前的范式，癌症起源于 DNA 水平的功能障碍。因此，研究 癌症的表观遗传调控历来集中于 DNA 甲基化、组蛋白的影响 修饰和染色质重塑（最后两个是 DNA 突变的结果）。然而，鉴于 研究人员发现，DNA 水平的突变特征不足以对许多癌症亚型进行分类 还开始探索 RNA 表观遗传学在癌症进展中可能发挥的作用。一种潜在的RNA 感兴趣的表观遗传机制是 RNA 编辑，或者通过以下方式对转录的 RNA 进行位点特异性修饰： 脱氨酶。已知 RNA 编辑会影响剪接、氨基酸编码和 microRNA (miRNA) 及其 目标。最近，出现的证据支持腺苷与肌苷（A-to- I) 癌症中的 RNA 编辑（Chen 等人，2013）。在这里，我试图确定是否胞嘧啶至尿嘧啶（C-to-U）编辑， 由 Apobec-1 介导，可在癌症进展中发挥致病作用。此前曾进行过基因实验 表明 Apobec-1 参与 Apcmin/+ 小鼠肠癌模型的疾病进展。 因此，我建议进行研究以了解 Apobec-1 介导的 RNA 编辑在 在这个肿瘤模型中，癌症进展的背景。我在初级哺乳动物细胞中的初步数据表明 C-to-U 编辑可以通过修改 miRNA 靶位点来影响编辑转录本的翻译水平。我 假设 Apobec-1 介导的 RNA 编辑通过以下方式促进肿瘤细胞更具侵袭性的表型 改变 miRNA 靶向来影响基因表达。这项工作将通过利用下一步来完成 世代测序、生物信息学和分子生物学技术。使用来自 Apcmin/+ 小鼠，我将： (a) 对整个转录组中的 C 到 U 编辑位点进行编目； (b) 识别潜在的 miRNA 受 Apobec-1 编辑影响的靶位点，重点关注潜在癌症相关基因中出现的靶位点；和 (c) 进行功能测定以评估已识别靶标的重要性。这些实验旨在 建立 Apobec-1 介导的 RNA 编辑与癌症之间的直接机制联系。此外，他们还将 还为癌症进展的新机制提供了证据，该机制可以扩展到所有癌症，并将 产生新的诊断工具和化疗靶点。 ！