Molecular Pathogenesis of Acute Myeloid Leukemia

急性髓系白血病的分子发病机制

基本信息

批准号：
10678908
负责人：
TIMOTHY J. LEY
金额：
$ 91.43万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-08-13 至 2029-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10678908
关键词：
ATAC-seq Acute Myelocytic Leukemia Back CBFA2T1 gene CBFbeta-MYH11 fusion protein Cell model Cells ChIP-seq Clinical Trials Design Communities Core-Binding Factor DNA DNA sequencing DNMT3B gene DNMT3a DNMT3a mutation Data Analyses Data Set Databases Development Disease Epigenetic Process Event Funding Genetically Engineered Mouse Genomics Goals Growth HDAC1 gene Hematopoietic Hematopoietic Neoplasms Hematopoietic stem cells Histones Human Methods Molecular Mutation PML-RARalpha protein Pathogenesis Patients Pharmaceutical Preparations Precision therapeutics Preleukemia Process Protein Isoforms Proteins Proteomics RUNX1 gene Sampling Series Work acute myeloid leukemia cell bisulfite sequencing blastomere structure embryo cell engineered stem cells epigenomics fitness genetic approach genomic locus induced pluripotent stem cell inhibitor insight leukemia initiating cell mouse model mutant novel overexpression phosphoproteomics programs proteogenomics single cell technology targeted treatment therapy design transcriptome sequencing whole genome

项目摘要

Project Summary The long-term goal of this project is to develop effective, precision therapies directed against the initiating mutations of Acute Myeloid Leukemia (AML). During the current funding period, we performed a series of genomic and epigenomic studies that have clarified the mechanisms that AML initiating mutations use to "reprogram" hematopoietic stem progenitor cells (HSPCs), increasing their fitness for transformation. In the next funding period, we will use primary human AML samples, induced pluripotent stem cells (iPSCs), and genetically engineered mouse models to further evaluate the molecular mechanisms involved in preleukemic reprogramming, and progression to AML. Four well characterized events (that initiate more than half of AML cases) will be studied in detail. We will continue our work with DNMT3A mutations and PML- RARA, and add the study of Core Binding Factor AML fusions (RUNX1-RUNX1T1 and CBFB-MYH11). The "toolkit" for these studies will involve the analysis of preleukemic and fully transformed hematopoietic cells from these models, using bulk DNA and RNA sequencing, whole genome bisulfite sequencing, ATAC-seq, ChIP- seq and/or CUT&RUN to detect the genomic locations of activating and repressive histone marks (and the fusions themselves), and single cell technologies for RNA, DNA, and ATAC-seq. We will also be performing comprehensive proteomic studies to complete "proteogenomic" datasets for these initiating events, including 1) the identification of the hematopoietic proteins that interact with the initiating proteins listed above, and 2) the development of quantitative deep-scale proteomic and phosphoproteomic datasets broadly representative of all AML subtypes. The integrative analysis of these datasets (and their availability to the AML community) should provide important new insights about AML pathogenesis, and suggest mechanistically targeted therapies. In this proposal, we provide one representative example of this process: using novel methods to identify proteins that interact with DNMT3A, we discovered several mutations that disrupt the normal interaction of DNMT3A with an inactive isoform of DNMT3B (DNMT3B3); these mutations destabilize DNMT3A and decrease its activity. Remarkably, we found that we can restore the activity of many mutant DNMT3A proteins by retrovirally overexpressing DNMT3L, a protein that normally interacts with DNMT3A and 3B in embryonic cells to increase their activity. "Addback" of DNMT3L into hematopoietic cells with the Dnmt3aR878H mutation remethylates DNA, and decreases the growth of AML cells initiated by this mutation. Since DNMT3L is epigenetically silenced in nearly all AMLs, a program to identify drugs and genetic strategies to reactivate DNMT3L in AML cells will be developed. We have already found that Romidepsin, an HDAC1 inhibitor, potently induces DNMT3L expression, and a clinical trial designed to evaluate the activity of this drug in DNMT3A mutant AMLs is planned. Additional approaches for developing mechanistically driven therapies designed to thwart initiating mutations will be developed during the next funding period.

项目摘要该项目的长期目标是开发针对该项目的有效的精确疗法引发急性髓样白血病（AML）的突变。在当前的资金期间，我们进行了一系列基因组和表观基因组研究阐明了AML启动突变使用的机制为了“重编程”造血干祖细胞（HSPC），增加了其转化的适应性。在下一个资金期，我们将使用原代人AML样品，诱导多能干细胞（IPSC），和基因工程的小鼠模型，以进一步评估涉及的分子机制 peleukemic重新编程，并发展为AML。四个特征性的事件（这会发起更多将详细研究比一半的AML病例）。我们将继续使用DNMT3A突变和PML- RARA，并添加核心结合因子AML融合的研究（RUNX1-RUNX1T1和CBFB-MYH11）。这对于这些研究这些模型，使用大量DNA和RNA测序，全基因组Bisulfite测序，ATAC-SEQ，芯片 - Seq和/或cut and cun and cun以检测激活和抑制性组蛋白标记的基因组位置（以及融合本身）以及用于RNA，DNA和ATAC-SEQ的单细胞技术。我们还将表演全面的蛋白质组学研究以完成这些启动事件的“蛋白质组学”数据集，包括1）鉴定与起始蛋白相互作用的造血蛋白上面列出，以及2）定量深度蛋白质组学和磷蛋白质组的发展数据集广泛代表所有AML亚型。这些数据集的集成分析（及其 AML社区的可用性）应提供有关AML发病机理的重要新见解，并提供建议机械靶向疗法。在此提案中，我们提供了一个代表性的例子过程：使用新型方法鉴定与DNMT3A相互作用的蛋白质，我们发现了几种突变这破坏了DNMT3A与DNMT3B无活性同工型的正常相互作用（DNMT3B3）；这些突变破坏了DNMT3A并减少其活性。值得注意的是，我们发现我们可以恢复活动通过倒流过表达DNMT3L的许多突变型DNMT3A蛋白（一种通常相互作用的蛋白）胚胎细胞中的DNMT3A和3B以增加其活性。 DNMT3L的“倒数”成造血具有DNMT3AR878H突变的细胞再甲基甲基DNA，并降低了由此引发的AML细胞的生长突变。由于DNMT3L在几乎所有AML中都表观遗传沉默，因此识别药物和遗传的程序将制定在AML细胞中重新激活DNMT3L的策略。我们已经发现Romidepsin，一个 HDAC1抑制剂有效诱导DNMT3L表达，并旨在评估活性计划在DNMT3A突变体AML中使用该药物。开发机械驱动的其他方法旨在阻止启动突变的疗法将在下一个资金期间开发。

项目成果

期刊论文数量（2）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Melanoma in a patient with DNMT3A overgrowth syndrome.

DOI：
10.1101/mcs.a006267
发表时间：
2023-04
期刊：
COLD SPRING HARBOR MOLECULAR CASE STUDIES
影响因子：
1.8
作者：
Chen, David Y.;Sutton, Leslie A.;Ramakrishnan, Sai Mukund;Duncavage, Eric J.;Heath, Sharon E.;Compton, Leigh A.;Miller, Christopher A.;Ley, Timothy J.
通讯作者：
Ley, Timothy J.