The role and mechanism of METTL3/METTL14-mediated RNA modification in the pathogenesis and drug-resistance of AML

METTL3/METTL14介导的RNA修饰在AML发病及耐药中的作用及机制

基本信息

批准号：
10558640
负责人：
Jianjun Chen
金额：
$ 45.09万
依托单位：
BECKMAN RESEARCH INSTITUTE/CITY OF HOPE
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-02-01 至 2024-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10558640
关键词：
11q23 Acute Myelocytic Leukemia Address Affect Anthracycline Ara-C Basic Science Biological Process Biology Bone Marrow Transplantation Cell Differentiation process Cells Chemoresistance Complex Coupled Cytarabine Cytogenetics Data Data Set Daunorubicin Deposition Development Disease Disease remission Drug resistance FLT3 gene Frequencies Genes Goals Hematopoietic Hematopoietic Neoplasms Hematopoietic stem cells Human In Vitro Knock-in Mouse Knockout Mice Leukemic Cell MLL-AF9 MLL-rearranged leukemia Maintenance Malignant Neoplasms Mediating Messenger RNA Methyltransferase Modeling Modification Molecular Mus Mutation NPM1 gene Neoadjuvant Therapy Oncogenic Pathogenesis Pathologic Pathway interactions Patients Play Post-Transcriptional Regulation Prognosis Proteins RNA RNA Stability Reader Refractory Regulation Relapse Reporting Research Design Resistance Role Stress Therapeutic Tissues Transgenic Organisms Translational Research Translations Treatment Failure Validation Xenograft Model Xenograft procedure acute myeloid leukemia cell bioprocess cancer type chemotherapy crosslinking and immunoprecipitation sequencing drug relapse gain of function in vivo insight leukemia leukemia treatment loss of function mouse model mutant novel novel therapeutic intervention novel therapeutics overexpression response ribosome profiling self-renewal side effect stem stem cell self renewal stem-like cell success transcriptome transcriptome sequencing validation studies virtual

项目摘要

PROJECT SUMMARY (ABSTRACT): Background: N6-methyladenosine (m6A) modification is the most abundant internal modification in eukaryotic messenger RNAs (mRNAs) and plays critical roles in various normal bioprocesses. Evidence is emerging that m6A and the associated machinery also play roles in cancers. Acute myeloid leukemia (AML) is one of the most common and fatal forms of hematopoietic malignancies with distinct cytogenetic (e.g., MLL-rearranged) and molecular (e.g., mutations in FLT3 and NPM1) abnormalities. Over 70% of AML patients cannot be cured by contemporary therapies due to drug resistance and relapse. The leukemia stem/initiating cells (LSCs/LICs) are considered to be the root cause for the treatment failure and relapse of AML. Thus, it is critical to better understand the molecular mechanisms underlying AML pathogenesis, LSC/LIC self-renewal and drug resistance, which may lead to the development of effective novel therapeutic strategies to treat AML. RNA m6A modification is deposited by the METTL3-METTL14 methyltransferase complex (MTC; i.e., writer), and may affect the stability and/or translation efficiency of target mRNAs. We and others have reported that both METTL3 and METTL14 play oncogenic roles in MLL-rearranged AML, but their definitive roles in the development and maintenance of other (i.e., non-MLL- rearranged) subtypes of AMLs and especially in LSC/LIC self-renewal remain elusive. Our unpublished preliminary data imply that METTL3 and METTL14 may also play roles in drug resistance in AML, which warrants further systematic studies. Furthermore, their critical targets in AMLs have yet to be well investigated. Hypothesis: METTL3 and METTL14 play essential oncogenic roles in AML pathogenesis, LSC/LIC maintenance and drug resistance through m6A-dependent regulation of expression of critical target genes. Specific Aims: 1) To determine the roles of METTL3 and METTL14 in the development and maintenance of different subtypes of AMLs, as well as in the self-renewal of relevant LSCs/LICs and normal HSCs; 2) To determine the roles of METTL3 and METTL14 in drug resistance of AMLs; and 3) To decipher the molecular mechanisms underlying the pathological roles of METTL3 and METTL14 in AML development/maintenance, LSC/LIC self-renewal and drug resistance. Study Design: 1) We will use inducible Mettl3 and Mettl14 knockout mouse models coupled with mouse AML models, as well as patient-derived xeno-transplantation (PDX) models, to investigate the roles of METTL3 and METTL14 in development/maintenance of different subtypes of AMLs and self-renewal of LSCs/ LICs and normal hematopoietic stem cells (HSCs). 2) We will conduct both gain- and loss-of-function in vivo studies to determine whether METTL3 and METTL14 play critical roles in drug resistance of AMLs. 3) We will perform PAR- CLIP-seq, m6A-seq, RNA-seq, RNA life-time profiling-seq, and ribosome profiling-seq to identify all direct targets of METTL3 and METTL14 in AMLs and to understand how METTL3 and METTL14 regulate expression of their target RNAs, followed by validation and functional studies of a set of top candidate targets.

项目概要（摘要）：背景：N6-甲基腺苷（m6A）修饰是最丰富的内部修饰真核生物信使 RNA (mRNA) 在各种正常生物过程中发挥着关键作用。证据是研究表明 m6A 和相关机制也在癌症中发挥作用。急性髓系白血病 (AML) 是最常见和致命的造血系统恶性肿瘤之一，具有独特的细胞遗传学（例如 MLL 重排）和分子（例如 FLT3 和 NPM1 突变）异常。超过 70% 的 AML 患者无法通过以下方法治愈由于耐药性和复发而导致的当代疗法。白血病干细胞/起始细胞 (LSC/LIC) 是被认为是AML治疗失败和复发的根本原因。因此，更好地理解这一点至关重要 AML发病机制、LSC/LIC自我更新和耐药性的分子机制，可能导致开发有效的新型治疗策略来治疗 AML。 RNA m6A 修饰由 METTL3-METTL14 甲基转移酶复合物（MTC；即 writer），可能会影响稳定性和/或翻译目标 mRNA 的效率。我们和其他人已经报道 METTL3 和 METTL14 都发挥致癌作用在MLL重排的AML中，但它们在其他（即非MLL- AML 的亚型，特别是 LSC/LIC 的自我更新，仍然难以捉摸。我们未发表的初步数据表明 METTL3 和 METTL14 也可能在 AML 耐药中发挥作用，这有理由进一步系统研究。此外，它们在反洗钱中的关键目标尚未得到充分研究。假设：METTL3 和 METTL14 在 AML 发病机制、LSC/LIC 中发挥重要的致癌作用通过 m6A 依赖性调节关键靶基因的表达来维持和耐药性。具体目标： 1）确定METTL3和METTL14在开发和维护中的作用不同亚型的 AML，以及相关 LSC/LIC 和正常 HSC 的自我更新； 2) 至确定 METTL3 和 METTL14 在 AML 耐药中的作用； 3）破译分子 METTL3 和 METTL14 在 AML 发展/维持中病理作用的潜在机制， LSC/LIC 自我更新和耐药性。研究设计： 1) 我们将使用可诱导的Mettl3和Mettl14敲除小鼠模型以及小鼠 AML 模型以及患者来源的异种移植 (PDX) 模型，以研究 METTL3 的作用和 METTL14 在 AML 不同亚型的开发/维护以及 LSC/LIC 的自我更新中的应用正常造血干细胞（HSC）。 2）我们将进行体内功能获得和丧失的研究确定 METTL3 和 METTL14 是否在 AML 耐药中发挥关键作用。 3）我们将执行PAR- CLIP-seq、m6A-seq、RNA-seq、RNA 寿命分析-seq 和核糖体分析-seq 可识别所有直接靶标 METTL3 和 METTL14 在 AML 中的作用，并了解 METTL3 和 METTL14 如何调节其表达目标 RNA，然后对一组顶级候选目标进行验证和功能研究。

项目成果

期刊论文数量（1）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

METTL16 promotes liver cancer stem cell self-renewal via controlling ribosome biogenesis and mRNA translation.

METTL16 通过控制核糖体生物发生和 mRNA 翻译促进肝癌干细胞自我更新。

DOI：
发表时间：
2024-02-01
期刊：
影响因子：
0
作者：
Xue, Meilin;Dong, Lei;Zhang, Honghai;Li, Yangchan;Qiu, Kangqiang;Zhao, Zhicong;Gao, Min;Han, Li;Chan, Anthony K N;Li, Wei;Leung, Keith;Wang, Kitty;Pokharel, Sheela Pangeni;Qing, Ying;Liu, Wei;Wang, Xueer;Ren, Lili;Bi, Hongjie;Yang, Lu;S
通讯作者：
S