Role of METTL3 and the m6A Epitranscriptome in cancer

METTL3 和 m6A 表观转录组在癌症中的作用

• 批准号: 10302284
• 负责人: Richard I. Gregory
• 金额: $ 63.36万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-12-11 至 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10302284
• 关键词: 5' Untranslated Regions; Address; Affect; Age; Apoptosis; Binding Sites; Biological Assay; CRISPR/Cas technology; Cancer Biology; Cancer Model; Cells; Cellular biology; Characteristics; Co-Immunoprecipitations; Complementary DNA; Complex; Data; Diagnosis; Enzymes; Fibroblasts; Future; Gene Expression; Genes; Genetic Transcription; Genotype; Goals; Human; Immunohistochemistry; In Vitro; Inhibition of Cancer Cell Growth; Investigation; KRASG12D; Lead; Lentivirus; Lentivirus Vector; Lung; Lung Adenocarcinoma; Lung Neoplasms; Malignant Neoplasms; Malignant neoplasm of lung; Maps; Mass Spectrum Analysis; Measures; Messenger RNA; Methods; Methyltransferase; Modeling; Modification; Molecular; Monitor; Mus; Normal tissue morphology; Oncogenes; Oncogenic; Organoids; Pathway interactions; Persons; Phosphorylation; Positioning Attribute; Primary Neoplasm; Proteins; Proteomics; RNA; RNA immunoprecipitation sequencing; RNA methylation; RNA, Messenger, Splicing; Regulation; Regulator Genes; Reporter; Research; Role; Sampling; Survival Rate; Terminator Codon; Testing; Time; Transgenic Organisms; Translations; Woman; Xenograft procedure; cohort; effective therapy; epitranscriptome; experience; experimental study; gain of function; in vivo; insight; knock-down; lung cancer cell; lung tumorigenesis; metaplastic cell transformation; mouse model; mutant; novel; novel therapeutic intervention; overexpression; protein expression; ribosome profiling; small hairpin RNA; therapeutic target; transcriptome; transcriptome sequencing; tumor; tumor growth; tumor initiation; tumor progression; tumorigenesis

Lung adenocarcinoma accounts for about 40% of all lung cancers and is the most common form of lung cancer found in women and in people under the age of forty-five. Strikingly, the 5-year survival rate for lung cancer overall is only about 18 percent, and more than half of people with lung cancer die within one year of diagnosis. This highlights the need for more effective treatment options and underscores the importance of research focused on uncovering and understanding new molecular and cellular pathways that contribute to lung cancer biology. Messenger RNAs (mRNAs) are subject to various posttranscriptional modifications including N6-Methyladenosine (m6A). m6A is the most abundant mRNA modification and is emerging as an important regulator of gene expression that can affect mRNA splicing, export, stability, and translation. m6A is catalyzed the METTL3 methyltransferase complex, and occurs at a characteristic sequence motif at a position close to the translation stop codon of a large subset of mRNAs. The goal of this proposal is to test the central hypothesis that METTL3 is a novel oncogenic factor in lung cancer. The global m6A mRNA `epitranscriptome' will be mapped and measured in a cohort of primary human lung tumor samples. Relative levels of METTL3 in tumors will be measured by immunohistochemistry and correlated with the m6A levels and transcriptome-wide distribution. Loss- and gain-of-function experiments will address the widespread impact of METTL3 (and METTL3-interacting proteins) in controlling target mRNA expression, and will help uncover the molecular and cellular role of METTL3, METTL3-interacting proteins, and a selection of downstream targets mRNAs, in lung cancer cell biology. Finally, the effects of METTL3 manipulation in lung tumor initiation and progression will be explored using a mouse lung cancer model, as well as a novel lung organoid model and a panel of assays will be deployed to examine the underlying molecular mechanism. Successful completion of the proposed studies will help establish METTL3 as a possible future therapeutic target for lung adenocarcinoma and other cancers.

肺腺癌约占所有肺癌的40%，是肺腺癌中最常见的一种。 女性和45岁以下人群中发现的癌症。引人注目的是，肺移植的5年存活率 肺癌的发病率只有18%，超过一半的肺癌患者在一年内死亡。 诊断.这突出了需要更有效的治疗方案，并强调了以下方面的重要性： 研究重点是发现和理解新的分子和细胞途径，有助于 肺癌生物学信使RNA（mRNA）受到各种转录后修饰 包括N6-甲基腺苷（m6 A）。m6 A是最丰富的mRNA修饰，正在成为一种新的基因修饰。 基因表达的重要调节因子，可影响mRNA剪接、输出、稳定性和翻译。m6A 被胃L3甲基转移酶复合物催化，并发生在一个特征序列基序上， 在一个大的mRNA亚群中靠近翻译终止密码子的位置。这项提案的目的是测试 核心假设是胃L3是肺癌中的一种新的致癌因子。全球m6 A mRNA 将在一组原发性人肺肿瘤样品中绘制和测量“表转录组”。相对 通过免疫组织化学测量肿瘤中的胃L3水平，并与m6 A水平相关联 和转录组分布。功能丧失和获得实验将解决广泛的 胃L3（和胃L3相互作用蛋白）在控制靶mRNA表达中的影响，并将有助于 揭示了胃L3的分子和细胞作用，胃L3相互作用蛋白，以及一系列 下游靶向mRNA，在肺癌细胞生物学中。最后，研究了针刺L3对肺的影响。 肿瘤的发生和发展将使用小鼠肺癌模型，以及一种新的肺 将部署类器官模型和一组测定来研究潜在的分子机制。 成功完成拟定的研究将有助于确立胃L3作为未来可能的治疗药物 靶向治疗肺腺癌和其他癌症。