Define the oncogenic role of METTL3 in the pathogenesis of chronic lymphocytic leukemia

定义 METTL3 在慢性淋巴细胞白血病发病机制中的致癌作用

• 批准号: 10717803
• 负责人: Lili Wang
• 金额: $ 55.62万
• 依托单位: BECKMAN RESEARCH INSTITUTE/CITY OF HOPE
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2028-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10717803
• 关键词: Address; Adoptive Transfer; Automobile Driving; B-Lymphocytes; Biology; CRISPR screen; Cell Line; Cell Proliferation; Cells; Chemotherapy-Oncologic Procedure; Chronic Lymphocytic Leukemia; Clinical; Clinical Trials; Country; Data; Data Set; Defect; Deposition; Development; Disease; Disease Progression; Essential Genes; Genes; Genetic Transcription; Growth; Hematologic Neoplasms; Hematopoietic stem cells; In Vitro; Interleukins; Knock-out; Knockout Mice; Knowledge; Maintenance; Malignant - descriptor; Malignant Neoplasms; Messenger RNA; Methyltransferase; Modeling; Modification; Molecular; Mus; Mutation; Oncogenes; Oncogenic; Oral; Outcome; Pathogenesis; Pathway interactions; Patients; Pharmacologic Substance; Play; Post-Transcriptional Regulation; Prognosis; Prognostic Marker; Proliferating; Proteins; Proteome; Proteomics; RNA; RNA Splicing; Regimen; Regulation; Reporting; Role; Sampling; Site; Spliceosomes; Technology; Transcript; Translations; Treatment Protocols; Up-Regulation; adult leukemia; cancer therapy; chemotherapy; chronic lymphocytic leukemia cell; design; experimental study; genome editing; genome-wide; improved; in vivo; independency; inhibitor; insight; mouse model; novel; novel therapeutic intervention; novel therapeutics; overexpression; posttranscriptional; predictive marker; protein complex; protein expression; ribosome profiling; small molecule inhibitor; targeted treatment; therapeutic target; therapeutically effective; transcription factor; transcriptome; transcriptome sequencing

Project Summary Aggressive chronic lymphocytic leukemia (CLL) remains incurable despite with improved chemotherapy regimens and targeted therapies. Better understanding of the biology underlying aggressive CLL is expected to design novel therapies. We recently discovered that high spliceosome complex protein expression results in aberrant RNA splicing, is associated with aggressive disease and serves as an independent predictive marker for poor prognosis in CLL, highlighting RNA splicing dysregulation underlies aggressive CLL. Through an integrated transcriptome and proteome analysis on normal and primary CLL B cells, we discovered that METTL3, an RNA methyltransferase that deposits N6-methyladenosine (m6A) modification on mRNA, is strongly implicated as a key regulator for RNA splicing in CLL. METTL3 is a highly expressed protein in CLL, associated with poor clinical outcome, and significantly correlated with splicing factor protein expression. Moreover, efficacious small molecule inhibitors of METTL3 have been developed and have shown promising results in hematological malignancy. Treatment with METTL3 inhibitor results in growth defect in CLL cell lines and decreases RNA splicing factor protein expression. Moreover, we have obtained evidence that METTL3 overexpression is oncogenic by rending IL-3 independency in Ba/F3 cell line model. Based on these preliminary data, we hypothesize that METTL3 potentially acts as an oncogene and drives the onset and progression of CLL through the regulation of RNA splicing network, making it a potential target for treating aggressive CLL. To address this hypothesis, we propose to investigate METTL3 is required for both development and maintenance of CLL using murine models (Aim 1). We will dissect the molecular mechanism underlying the targets of RNA splicing network in CLL and understand how they are regulated by METTL3 (Aim 2). Moreover, we will determine whether METTL3 is a viable therapeutic target in treating aggressive CLL and if targeting both METTL3 and RNA splicing regulatory network has a synergistic effect in CLL (Aim 3). Collectively, the results of the experiments proposed in this application will establish METTL3 as an CLL oncogene, elucidate the m6A-modification on target transcripts that modulated by METTL3 to promote and maintain CLL, and evaluate a new therapeutic approach for aggressive CLL.

项目摘要 侵袭性慢性淋巴细胞白血病（CLL）尽管化疗得到改善，但仍然无法治愈 方案和靶向治疗。更好地理解侵袭性CLL的生物学基础， 设计新的疗法。 我们最近发现，高剪接体复合蛋白表达导致RNA剪接异常， 与侵袭性疾病相关，并作为CLL预后不良的独立预测标志物， 突出RNA剪接失调是侵袭性CLL的基础。通过整合的转录组， 通过对正常和原代CLL B细胞的蛋白质组分析，我们发现胃L3，一种RNA甲基转移酶， 在mRNA上沉积N6-甲基腺苷（m6 A）修饰，强烈暗示是RNA的关键调节因子 CLL中的剪接。CLL中L3是一种高表达的蛋白质，与临床结局差相关， 与剪接因子蛋白表达显著相关。此外，有效的小分子抑制剂， 已经开发了胃L3，并在血液恶性肿瘤中显示出有希望的结果。治疗 胃L3抑制剂导致CLL细胞系的生长缺陷并降低RNA剪接因子蛋白表达。 此外，我们已经获得证据表明，胃L3过表达是通过使IL-3非依赖性致癌的， 在Ba/F3细胞系模型中。基于这些初步数据，我们假设胃L3可能作为一种 癌基因，并通过调节RNA剪接网络驱动CLL的发作和进展， 它是治疗侵袭性CLL的潜在靶点。为了解决这一假设，我们建议研究胃L3 是使用鼠模型发展和维持CLL所必需的（Aim 1）。我们将解剖 CLL中RNA剪接网络靶点的分子机制，并了解它们是如何 受AAPL 3（目标2）监管。此外，我们将确定胃L3是否是一个可行的治疗靶点， 治疗侵袭性CLL，如果靶向胃L3和RNA剪接调控网络， 对慢性淋巴细胞白血病（CLL）的影响（目标3）。 总的来说，本申请中提出的实验的结果将确立胃L3作为CLL 癌基因，阐明m6 A修饰的目标转录本，调节胃L3，以促进和 维持CLL，并评估侵袭性CLL的新治疗方法。