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

描述（由申请人提供）：根据目前的范式，癌症起源于DNA水平的功能障碍。因此，对癌症表观遗传调控的研究历来集中在DNA甲基化、组蛋白修饰和染色质重塑的影响上（后两者是DNA突变的结果）。然而，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。一个潜在的RNA表观遗传机制是RNA编辑，或通过脱氨酶对转录的RNA进行位点特异性修饰。众所周知，RNA编辑会影响剪接、氨基酸编码、microrna （mirna）及其靶标。最近，有证据支持腺苷到肌苷（a -to- I） RNA编辑在癌症中的直接致病作用（Chen et al., 2013）。

DESCRIPTION (provided by applicant): 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.