The Novel Role and Mechanism of RBM33 in Leukemogenesis

RBM33 在白血病发生中的新作用和机制

基本信息

批准号：
10623163
负责人：
Zhijian Qian
金额：
$ 39.79万
依托单位：
UNIVERSITY OF FLORIDA
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-05-17 至 2027-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10623163
关键词：
Acute Myelocytic Leukemia Binding Binding Sites Biological Process Cells Complex DNA Sequence Rearrangement Data Dependence Development Ensure Essential Genes Gene Expression Gene Expression Regulation Gene Mutation Genes Genetic Models Genomic approach Growth Hematologic Neoplasms Hematopoiesis Hematopoietic stem cells Human Leukemic Cell Machine Learning Maintenance Maps Mediating Messenger RNA Methylation Modification Molecular Mus Oncogenes Pathogenesis Pathogenicity Pathway interactions Patients Play Polyribosomes Post-Transcriptional Regulation Protein Binding Domain Proteomics RNA RNA Binding RNA methylation RNA-Binding Proteins Reader Regulation Ribosomes Role Site TACC3 gene Testing Transcript crosslinking and immunoprecipitation sequencing demethylation epitranscriptome genomic data improved in vivo knock-down leukemia leukemia treatment leukemic stem cell leukemogenesis member mouse genetics mouse model novel novel therapeutic intervention posttranscriptional recruit self-renewal stem stem cell self renewal success targeted treatment transcriptome transcriptome sequencing

项目摘要

Abstract Acute myeloid leukemia (AML) is one of the most common and fatal forms of hematological malignancies caused by gene mutations and genomic rearrangements. The cure rates for AML patients have not significantly improved for decades. The molecular mechanisms underlying the pathogenesis of AML are not fully understood. By analysis of publicly available genomic data using a new machine learning approach, RNA Binding Motif Protein 33 (RBM33), an RNA binding protein, is identified as an essential gene in AML. However, the biological function of RBM33 is unknown yet. Our preliminary studies provide the first compelling evidence suggesting a novel function of RBM33 in regulating m6A RNA demethylation. More importantly, we showed that RBM33 knockdown significantly inhibited growth and survival of human and mouse leukemia cells. At a molecular level, we have identified a potential downstream target of RBM33 in leukemia cells. ALKBH5 is known as an m6A mRNA demethylase (Eraser), which removes m6A methylated groups from RNA. To date, it remains unknown whether another member of RNA binding proteins is required for ensuring recruitment of ALKBH5 to its mRNA targets. We have recently demonstrated that ALKBH5 has a critical role in AML development and maintenance. We hypothesize that RBM33 plays a critical role in the pathogenesis of AML by regulating ALKBH5-mediated m6A demethylation. To test this hypothesis, we will pursue three specific aims. In Aim1, we will determine a novel role of RBM33 in regulation of dynamic m6A RNA methylation in leukemia cells. In Aim2, we will investigate the role of Rbm33 in AML development and maintenance. In Aim 3, we will determine the downstream pathway that mediates the function of RBM33 in leukemogenesis in AML. We will employ both mouse genetic models as well as human patient-derived mouse models to elucidate the role of RBM33 in normal hematopoiesis and leukemogenesis in vivo, and will combine transcriptome and epitranscriptome analysis to identify the key downstream targets and associated downstream pathways that mediate the role of RBM33 in leukemogenesis. Our studies will uncover a novel role of RBM33 in m6A RNA modification, and define the importance and underlying mechanisms of RBM33 in AML development and maintenance as well as LSC/LIC self-renewal. Thus, the success of our project will significantly advance our understanding of the complex mechanisms underlying the m6A modification-mediated gene regulation in leukemia cells and the critical role of m6A RNA demethylation in leukemogenesis.

抽象的急性髓系白血病 (AML) 是最常见和致命的血液恶性肿瘤之一由基因突变和基因组重排引起。 AML患者的治愈率尚未达到几十年来显着改善。 AML发病机制的分子机制尚不明确完全明白了。通过使用新的机器学习方法分析公开可用的基因组数据，RNA 结合基序蛋白 33 (RBM33) 是一种 RNA 结合蛋白，被确定为 AML 中的必需基因。然而，RBM33的生物学功能尚不清楚。我们的初步研究提供了第一个令人信服的有证据表明 RBM33 在调节 m6A RNA 去甲基化方面具有新功能。更重要的是，我们表明 RBM33 敲低显着抑制人类和小鼠白血病的生长和存活细胞。在分子水平上，我们已经确定了白血病细胞中 RBM33 的潜在下游靶点。 ALKBH5 被称为 m6A mRNA 去甲基化酶（Eraser），可从 RNA 中去除 m6A 甲基化基团。迄今为止，尚不清楚是否需要 RNA 结合蛋白的另一个成员来确保将 ALKBH5 招募到其 mRNA 靶点。我们最近证明 ALKBH5 在 AML 开发和维护。我们假设 RBM33 在疾病的发病机制中发挥着关键作用通过调节 ALKBH5 介导的 m6A 去甲基化来治疗 AML。为了检验这个假设，我们将追求三个具体的目标。在 Aim1 中，我们将确定 RBM33 在调节动态 m6A RNA 甲基化中的新作用白血病细胞。在 Aim2 中，我们将研究 Rbm33 在 AML 开发和维护中的作用。在目标 3 中，我们将确定介导 RBM33 在 AML 白血病发生中的功能的下游途径。我们将采用小鼠遗传模型以及人类患者衍生的小鼠模型来阐明 RBM33 在体内正常造血和白血病发生中的作用，并将转录组和表观转录组分析以确定关键的下游靶点和相关的下游途径介导 RBM33 在白血病发生中的作用。我们的研究将揭示 RBM33 在 m6A RNA 中的新作用修改，并定义 RBM33 在 AML 发展和中的重要性和潜在机制维护以及 LSC/LIC 自我更新。因此，我们项目的成功将极大地推进我们的了解 m6A 修饰介导的基因调控的复杂机制白血病细胞和 m6A RNA 去甲基化在白血病发生中的关键作用。