PO.MCB06.03 · 分子与细胞生物学
RNA 修饰 m6A 的失调促进癌症进展并提供治疗机会
Dysregulation of the RNA modification m6A promotes cancer progression and provides therapeutic opportunities
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
本研究的目的是确定对 RNA 修饰 N6-甲基腺苷（m6A）的敏感检测和定量如何能够实现癌症检测、对肿瘤生物学的机制性洞见以及对治疗反应的改进监测。RNA 修饰是对核碱基或核糖的共转录和转录后改变，是几乎所有 RNA 代谢方面的关键调控因子，包括转录、剪接、定位、翻译、稳定性以及与 RNA 结合蛋白的相互作用。其中，m6A 是人类 mRNA 和长非编码 RNA 中最丰富的修饰，并在癌症中频繁失调。为实现对 RNA 修饰的全面分析，我们应用了一种多重邻近条形码检测，该检测富集修饰的 RNA 片段，在 cDNA 合成过程中编码修饰位点，并使用合成掺入对照进行校准以量化其在每个位点的丰度。该方法允许同时检测 m6A、肌苷和假尿苷，提供对癌症表观转录组的多维、高敏感性视图。将该检测应用于神经母细胞瘤样本和细胞系来源的异种移植瘤模型，揭示了在所检测的所有肿瘤类型中 m6A 丰度的一致性增加。值得注意的是，m6A 峰模式还区分了 MYCN 扩增型（MNA⁺）与 ALT 激活型（ALT⁺）和低风险神经母细胞瘤，表明该方法能够解析生物学和临床上不同的亚组。用 STC-15（一种靶向主要 m6A 写入酶 METTL3 的小分子抑制剂）处理神经母细胞瘤细胞系和异种移植瘤来源样本，导致该检测可测量的 m6A 水平呈剂量依赖性降低。METTL3 抑制与肿瘤细胞活力降低以及对化疗药物（如多柔比星）敏感性增加相关。对 m6A 及相关 RNA 修饰的敏感、定量测量提供了一种强大的策略，用于检测癌症相关的表观转录组失调、剖析神经母细胞瘤的分子异质性，以及监测对 METTL3 靶向干预的治疗反应。
查看英文原文 English abstract
The objective of this study is to determine how sensitive detection and quantitation of the RNA modification N 6-methyladenosine (m6A) can enable cancer detection, mechanistic insights into tumor biology, and improved monitoring of treatment responses. RNA modifications, which are co- and post-transcriptional alterations to nucleobases or ribose, are key regulators of nearly all aspects of RNA metabolism, including transcription, splicing, localization, translation, stability, and interactions with RNA-binding proteins. Among these, m6A is the most abundant modification in human mRNA and long noncoding RNA and is frequently dysregulated in cancer. To enable comprehensive analysis of RNA modifications, we applied a multiplexed proximity-barcoding assay that enriches for modified RNA fragments, encodes modification sites during cDNA synthesis, and quantifies their abundance at each locus using synthetic spike-in controls for calibration. This approach allows concurrent detection of m6A, inosine, and pseudouridine, providing a multidimensional, high-sensitivity view of the cancer epitranscriptome. Application of this assay to neuroblastoma samples and cell line-derived xenograft tumor models revealed consistent increases in m6A abundance across all tumor types examined. Notably, m6A peak patterns also distinguished MYCN-amplified (MNA⁺) from ALT-activated (ALT⁺) and low-risk neuroblastomas, demonstrating that the method can resolve biologically and clinically distinct subgroups. Treatment of neuroblastoma cell lines and xenograft-derived samples with STC-15, a small-molecule inhibitor targeting the primary m6A writer enzyme METTL3, led to a dose-dependent reduction in m6A levels measurable by this assay. METTL3 inhibition was associated with decreased tumor cell viability and increased sensitivity to chemotherapeutics such as doxorubicin. Sensitive, quantitative measurement of m6A and related RNA modifications provides a powerful strategy to detect cancer-associated epitranscriptomic dysregulation, to dissect molecular heterogeneity in neuroblastoma, and to monitor therapeutic responses to METTL3-targeted interventions.
利益披露 Disclosure
G. Stengel, None..
Z. Miles, None..
E. Davis, None..
B. Purse, None.