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. !

项目摘要 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在DNA甲基化、组蛋白 修饰和染色质重塑（最后两个是DNA突变的结果）。但鉴于 DNA水平上的突变特征不足以对许多癌症亚型进行分类，研究人员 也开始探索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，或转录RNA的位点特异性修饰， 脱氨酶。已知RNA编辑影响剪接、氨基酸编码和微RNA（miRNA）及其表达。 目标的最近，有证据表明腺苷对肌苷（A-对-B-）的直接致病作用。 I）癌症中的RNA编辑（Chen等人，2013年）。在这里，我试图确定胞嘧啶到尿嘧啶（C到U）编辑， 由Apobec-1介导，可以在癌症进展中发挥致病作用。基因实验以前 表明Apobec-1参与了肠癌的Apcmin/+小鼠模型中的疾病进展。 因此，我建议进行研究，以了解Apobec-1介导的RNA编辑在细胞凋亡中的机制作用。 在这个肿瘤模型中，癌症进展的背景下。我在原代哺乳动物细胞中的初步数据表明 C-to-U编辑可以通过修饰miRNA靶位点来影响编辑转录物的翻译水平。我 假设Apobec-1介导的RNA编辑促进肿瘤细胞中更具侵袭性的表型， 改变miRNA靶向以影响基因表达。这项工作将通过开发下一个 代测序、生物信息学和分子生物学技术。利用小肠组织 Apcmin/+小鼠，我将：（a）在整个转录组中对C到U编辑位点进行编目;（B）鉴定潜在的miRNA 受Apobec-1编辑影响的靶向位点，重点关注那些发生在潜在癌症相关基因中的位点; (c)进行功能测定以评估所鉴定的靶标的重要性。这些实验旨在 在Apobec-1介导的RNA编辑和癌症之间建立直接的机制联系。此外，他们将 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，并且将 产生新的诊断工具和化疗靶点。 !