Elucidating the role of mRNA m6A methylation in tumorigenesis

项目摘要/摘要 目前缺乏有效的、有针对性的癌症治疗方法，反映了人们对 促进肿瘤发生的机制，并强调需要新的治疗方法。为 几十年来，遗传驱动因素被认为是促进肿瘤发生的唯一机制。近期 研究进展建立了表位转录组学，它详细描述了RNA的调节和功能作用 修饰，作为一种新的肿瘤发生机制和潜在的治疗途径。N6- 甲基腺苷(M6A)是信使RNA(Mrna)最常见的修饰形式，在 基因调控和表达。然而，我们对mRNAm6A甲基化如何有助于 肿瘤的发生是有限的，需要进一步的研究。我的提案的F99阶段将决定 Fe~(2+)/𝛼𝛼-酮戊二酸依赖的双加氧酶碱性B家族成员ALKBH1在细胞周期中的新作用 砷致皮肤肿瘤中m6A基因甲基化的调控。长期接触无机物 受污染的饮用水中的砷是皮肤癌的主要致癌因素。然而， 砷致皮肤肿瘤的机制目前仍知之甚少，治疗也很少。 目标已经确定。我推测ALKBH1通过以下途径促进砷诱导的皮肤肿瘤发生 使m6A在mRNA上去甲基化，并在转录后调节基因表达。这方面的F99阶段 该提案将解决围绕砷分子机制的关键知识空白-- 诱导皮肤致瘤性及建立ALKBH1和/或其靶点作为砷中毒的治疗靶点 皮肤癌。K00阶段将解决一个更广泛的问题，并描述m6A甲基化在 恶变。恶变是一个动态的过程。然而，我们对 推动恶性转化的机制仅限于非转化和非转化的静态比较 转化的细胞和调节这些细胞类型之间转化的过渡状态仍然存在 没有特征的。K00阶段将通过使用m6A纵向测序来解决知识中的这一差距 使用一组经过验证的体外转化模型描述依赖m6A的表位转录组学 癌症。我假设依赖于m6A的表位转录组会在整个过程中动态变化 我将确定新的M6A依赖靶点，这些靶点将增加我们的 理解从正常细胞到致瘤细胞的转化过程。 成功完成该提案的F99和K00阶段将建立M6A的新机制 调节和功能，并确定新的治疗靶点和潜在的癌症发展生物标志物。 我的最终职业目标是成为一名独立的调查员，并管理一个跨学科的实验室 由美国国立卫生研究院资助的研究表位转录和癌症之间相互作用的工作。

PROJECT SUMMARY/ABSTRACT The current lack of effective, targeted cancer therapies reflects a gap in knowledge surrounding the mechanisms that promote tumorigenesis and emphasizes the need for new therapeutic approaches. For decades, genetic drivers were thought to be the sole mechanism that promote tumorigenesis. Recent advances have established epitranscriptomics, which details the regulatory and functional roles of RNA modifications, as an emerging mechanism of tumorigenesis and potential therapeutic avenue. N6- methyladenosine (m6A) is the most prevalent modification on messenger RNA (mRNA) and serves key roles in gene regulation and expression. However, our understanding of how mRNA m6A methylation contributes to tumorigenesis is limited and warrants further investigation. The F99 phase of my proposal will determine the novel role of ALKBH1, a member of the AlkB family of Fe2+/𝛼𝛼-ketoglutarate-dependent dioxygenases, in regulating mRNA m6A methylation in arsenic-induced skin tumorigenicity. Chronic exposure to inorganic arsenic through contaminated drinking water is a major carcinogenic driver of skin cancer. However, the mechanisms that underlie arsenic-induced skin tumorigenicity remain poorly understood and few therapeutic targets have been identified. I hypothesize that ALKBH1 promotes arsenic-induced skin tumorigenesis by demethylating m6A on mRNA and regulating gene expression post-transcriptionally. The F99 phase of this proposal will address critical gaps in knowledge surrounding the molecular mechanisms that underlie arsenic- induced skin tumorigenicity and establish ALKBH1 and/or its targets as therapeutic targets for arsenic-induced skin cancer. The K00 phase will address a broader question and profile the dynamic role of m6A methylation in malignant transformation. Malignant transformation is a dynamic process. However, our understanding of the mechanisms that drive malignant transformation is limited to static comparisons across non-transformed and transformed cells and the transitional states that mediate the transformation between these cell types remain uncharacterized. The K00 phase will address this gap in knowledge by using m6A-sequencing to longitudinally profile the m6A-dependent epitranscriptome using validated models of in vitro transformation across a panel of carcinomas. I hypothesize that the m6A-dependent epitranscriptome changes dynamically throughout malignant transformation and that I will identify novel m6A-dependent targets that will increase our understanding of the processes that underlie the transformation from a normal cell to a tumorigenic cell. Successful completion of the F99 and K00 phases of this proposal will establish novel mechanisms of m6A regulation and function and identify new therapeutic targets and potential biomarkers of cancer development. My ultimate career goal is to become an independent investigator and run an interdisciplinary lab conducting NIH-funded work that investigates the interplay between epitranscriptomics and cancer.