Role of m6A RNA methylation in Regulation of Translation in Human Glioblastoma

摘要 胶质母细胞瘤是最具侵袭性的人类脑癌类型，预后很差。近期 研究表明，肿瘤内转录表达存在广泛的异质性，并增加了肿瘤的可塑性 人类胶质瘤干细胞，被认为是治疗失败和肿瘤复发的原因。的作用 M6A RNA和ALKBH5在脑胶质瘤干细胞自我更新和致瘤性维持中的作用 最近已经被证明了。关于m6A RNA甲基化在体内的作用，目前知之甚少。 人类癌细胞翻译效率的调节。此外，m6A RNA甲基化的作用 包括m6A“写入器”(甲基转移酶，如Mettl14、METTL3)和“擦除器”(去甲基酶)的机械 例如ALKBH5，FTO)，在人脑胶质瘤干细胞向 分化的胶质瘤细胞尚未被研究。 我们假设，m6A RNA甲基化水平的改变会影响某些特定基因的翻译速度 人脑胶质瘤干细胞向分化细胞转化过程中的转录。我们建议M6A的损失 RNA甲基化提高了带有m6A峰区序列基序的转录本的翻译速度 它们与特定miRNAs的种子序列互补。这些miRNAs相互作用并调节 人脑胶质瘤细胞中的M6A RNA机制。 我们建议的具体目标将提供对M6A角色的全面和亚型特定洞察 RNA在hGSC和分化的GC之间的翻译调节中的作用： -SA1：进行RNA测序、MERIP测序、核糖体测序和计算分析 额外的人脑胶质瘤干细胞和分化的脑胶质瘤细胞 -SA2：确定m6A RNA甲基化峰内的miRNA结合的功能意义和 MiRNAs如何调控RNA甲基化机制。 我们的建议是对m6A RNA甲基化在调节 HGSCs向分化细胞转化过程中的翻译。此外，它还介绍了特定的角色 通过调节RNA甲基化机制，miRNAs作为HGSC翻译的调节者。自.以来 HGSCs向分化细胞的可逆性转变是肿瘤复发的关键因素 攻击性，识别这一过程的分子调控因子可能为治疗提供新的靶点 干预措施。

Abstract Glioblastoma represents the most aggressive type of human brain cancer with dismal prognosis. Recent studies have revealed extensive intratumoral heterogeneity in transcript expression and increased plasticity of human glioma stem cells, which are thought to contribute to treatment failure and tumor recurrence. The role of m6A RNA and ALKBH5 in the regulation of self-renewal and maintenance of tumorigenicity of glioma stem cells has been recently demonstrated. Limited knowledge exists with regards to the role of m6A RNA methylation in the regulation of translation efficiency in human cancer cells. Moreover, the role of m6A RNA methylation machinery that includes m6A “writers” (methyltransferases e.g. Mettl14, Mettl3) and “erasers” (demethylases e.g. ALKBH5, FTO), in the regulation of translation during the transition of human glioma stem cells to differentiated glioma cells has not been studied. We hypothesize that alterations at the level of m6A RNA methylation influences the rate of translation of certain transcripts during the transition of human glioma stem cells to differentiated cells. We propose that loss of m6A RNA methylation increases the rate of translation of transcripts with sequence motifs of their m6A peak regions that are complementary to the seed sequences of specific miRNAs. These miRNAs interact and regulate the m6A RNA machinery in human glioma cells. The specific aims of our proposal will provide a comprehensive and subtype specific insight into the role of m6A RNA in the regulation of translation during the transition between hGSCs and differentiated GCs: -SA1: Perform RNA sequencing, MeRIP sequencing, Ribo-sequencing and computational analysis of 10 additional human glioma stem cells and differentiated glioma cells -SA2: Determine the functional significance of the miRNA binding within the m6A RNA methylation peaks and how the miRNAs regulate RNA methylation machinery. Our proposal constitutes a functional exploration of the role of m6A RNA methylation in the regulation of translation during the transition of hGSCs to differentiated cells. In addition, it introduces the role of specific miRNAs as regulators of hGSC translation through modulation of the RNA methylation machinery. Since the reversible transition of hGSCs to differentiated cells constitutes a critical denominator of tumor recurrence and aggression, identification of molecular regulators of this process could provide novel targets for therapeutic interventions.