Role of m6A mRNA methylation in Neuroblastoma

M6A基因甲基化在神经母细胞瘤中的作用 过去30年的研究努力提供了表观遗传失调的广泛证据 癌症，其中DNA甲基化在肿瘤的生长和生存中起着关键作用。改变DNA的药物 通过抑制DNA甲基转移酶(MTase)的甲基化被用于临床治疗各种恶性肿瘤。 研究发现，哺乳动物信使RNA(MRNAs)也受到各种转录后转录的影响 包括m6A甲基化在内的修饰。我们的目标是绘制N6-甲基腺苷(M6A)的“表位转录组” 探讨神经母细胞瘤在儿童侵袭性肿瘤中的分子和细胞作用。 Nb细胞中的M6A甲基转移酶METTL3。使用已建立的协议，我们将在整个 人神经母细胞系的转录组，并将该方法并行地应用于分析原代 在应用优化方法研究m6A谱之前，来自明确定义的NB小鼠模型的肿瘤 在人类神经母细胞瘤标本中。我们对已公布的微阵列数据集的初步检查显示，METTL3 在一组NB肿瘤中，mRNA表达升高，且METTL3的高表达显著 与较差的患者存活率有关。我们将首次测量METTL3蛋白在肿瘤中的表达。 将研究METTL3在人类NB细胞中的分子和细胞作用，以表征可能的 METTL3在鼻咽癌细胞生物学中的致癌作用这项工作将在全基因组范围内提供第一个转录 任何肿瘤类型的m6A图谱，将在全球范围内识别直接的METTL3靶向信使RNA(MRNAs)，发现 M6A和METTL3在控制mRNA翻译方面的广泛影响，并将使METTL3成为一种 可能是未来治疗NB和其他癌症的治疗靶点。总体而言，这一创新提议 将建立分析肿瘤中m6A表位转录组的方法，并将对 确定表位转录组学领域对了解癌症生物学的重要性。

Role of m6A mRNA methylation in Neuroblastoma Research efforts over the last thirty years have provided widespread evidence of epigenetic dysregulation in cancer, in which DNA methylation plays a critical role in tumor growth and survival. Drugs that alter DNA methylation by inhibiting the DNA methyltransferases (MTase) are used clinically to treat various malignancies. It is emerging that mammalian messenger RNAs (mRNAs) are also subject to various posttranscriptional modifications including m6A methylation. Our goal is to map the N6-methyladenosine (m6A) ‘epitranscriptome’ in the aggressive pediatric cancer neuroblastoma (NB) and to explore the molecular and cellular role of the m6A methyltransferase METTL3 in NB cells. Using an established protocol we will map m6A throughout the transcriptome of human NB cell lines, and in parallel adapt and apply the approach to the analysis of primary tumors from a well-defined NB mouse model before applying the optimized method to investigate m6A profiles in human NB tumor samples. Our preliminary inspection of published microarray datasets reveal that METTL3 mRNA expression is elevated in a subset of NB tumors and that high METTL3 expression significantly correlates with poor patient survival. We will measure for the first time METTL3 protein expression in tumors. The molecular and cellular role of METTL3 in human NB cells will be investigated to characterize the possible oncogenic function of METTL3 in NB cancer cell biology. This work will provide the first transcriptome-wide map of m6A in any tumor type, will globally identify direct METTL3 target messenger RNAs (mRNAs), uncover the widespread impact of m6A and METTL3 in controlling mRNA translation, and will establish METTL3 as a possible therapeutic target for the future treatment of NB and other cancers. Overall, this innovative proposal will establish methods for analyzing the m6A epitranscriptome in tumors and will have a major impact on determining the importance of the field of epitranscriptomics to understanding cancer biology.