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Determining the Role of DNA Methylation Deregulation in Oncogenesis

确定 DNA 甲基化失调在肿瘤发生中的作用

基本信息

批准号：
10132255
负责人：
G Greg Wang
金额：
$ 34.86万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-04-01 至 2023-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10132255
关键词：
Acetylation Acute Acute Myelocytic Leukemia Affect Animal Model Animals Apoptosis Binding Binding Sites CRISPR/Cas technology Cancer Patient Cancerous Cell Line Cell Proliferation Cells Cellular Stress Chemicals Clinic Clinical Collection Complex Cytosine DNA DNA Methylation DNA Modification Methylases DNA Sequence Alteration Data Development Enhancers Enzymes Epigenetic Process Event FLT3 gene Gene Activation Gene Expression Genes Genomics Goals Growth Histone Acetylation Histones Human Hypersensitivity In Vitro Induced Mutation Knock-out MEIS1 gene MLLT3 gene Malignant - descriptor Malignant Neoplasms Mediating Methylation Methyltransferase Modeling Molecular Mus Mutate Mutation Myelogenous Myeloid Cells Oncogene Deregulation Outcome Paper Pathogenesis Pathway interactions Patients Phenotype Phosphotransferases Prognosis RUNX1 gene Recurrence Regulation Reporting Research Role Sampling Signal Pathway Site Somatic Mutation Survival Rate Technology Testing Therapeutic Transcription Elongation Translating base cancer cell cancer therapy cell transformation epigenome epigenomics hematopoietic stem cell self-renewal histone methyltransferase human disease improved in vivo inhibitor/antagonist innovation insight knock-down leukemia loss of function mutant neoplastic cell novel novel strategies patient derived xenograft model programs recruit screening self-renewal senescence small molecule inhibitor stem cell self renewal transcription factor transcriptome treatment strategy tumor tumorigenesis

项目摘要

PROJECT SUMMARY/ABSTRACT Somatic mutation of DNA (cytosine-5)-methyltransferase 3A (DNMT3AMut) occurs in >20-30% of acute myeloid (AML) patients making it one of the most frequently mutated genes in this deadly human disease. DNMT3AMut correlates with poor clinical outcome of AML. However, the molecular mechanism by which DNMT3AMut contributes to AML development remains far from clear. We reported that DNMT3AMut promotes cell transformation and establishes a bona fide AML phenotype in mice possessing the initiating RAS mutation. Integrated transcriptome and epigenomic profiling of this new murine AML model revealed that DNMT3AMut binds directly to enhancers of target genes inducing focal DNA hypo-methylation at binding sites. These DNMT3AMut-mediated events result in aberrant activation of a gene-expression program controlling stem cell self-renewal (notably a Meis1 node), anti-differentiation and tumor cell pro-survival. Our discovery-based epigenetic inhibitor screen further identified the DOT1L histone methyltransferase to be essential for DNMT3AMut-induced aberrant gene activation. We hypothesize that DNMT3AMut induces focal DNA hypo- leukemia methylation at gene enhancers promoting DOT1L-dependent activation of self-renewal (notably Meis1), anti- differentiation and tumor pro-survival genes, which collectively contribute to AML pathogenesis. Dissecting the molecular events and mechanism underlying DNMT3AMut -mediated AML progression should provide critical insights into new treatment strategies. Towards this goal, we will use cutting-edge CRISPR/Cas9 technologies to define the causal role for enhancer DNA hypomethylation due to DNMT3AMut in inducing aberrant gene activation and promoting AML determine what DNMT3AMut-activated gene pathways are essential for AML development in mice in vitro and in vivo (aim 1); we will through loss-of-function studies in our established murine AML models (aim 2); and third, in a translational aim, we will use human AML cell lines and primary patient-derived xenograft (PDX) models bearing DNMT3AMut to define effects of DNMT3AMut on epigenomic alteration, gene expression deregulation and malignant growth in human AML (aim 3). We expect to define the DNMT3AMut-induced epigenetic/gene changes in AML and expect to identify the pathway by which DNMT3AMut promotes AML progression. Because certain identified pathways such as DOT1L and Bcl2 are potentially druggable with the existing compounds , completion of our proposed research should not only promote a new mechanistic understanding of DNMT3AMut-associated AML but will also yield innovative therapeutics for the treatment of affected cancer patients.

项目摘要/摘要 DNA(胞嘧啶-5)-甲基转移酶3A(DNMT3AMut)的体细胞突变发生在20-30%的急性髓系白血病 (AML)患者使其成为这种致命人类疾病中最频繁的突变基因之一。 DNMT3AMut与急性髓系白血病的临床转归相关。然而，其分子机制 DNMT3AMut对AML发展的贡献仍远不清楚。我们报道了DNMT3AMut促进细胞转化，并在具有启动RAS突变的小鼠中建立真正的AML表型。对这一新的小鼠AML模型的整合转录组和表观基因组图谱显示DNMT3AMut 直接与靶基因的增强子结合，在结合部位引起DNA的局部低甲基化。这些 DNMT3AMut介导的事件导致控制干细胞的基因表达程序异常激活自我更新(特别是MEIS1结节)、抗分化和促进肿瘤细胞存活。我们基于发现的表观遗传抑制物筛选进一步鉴定DOT1L组蛋白甲基转移酶是 DNMT3AMut诱导的基因异常激活。我们推测DNMT3AMut可诱导局灶性DNA缺失。白血病基因增强子甲基化促进DOT1L依赖的自我更新激活(特别是Meis1)，反式分化和肿瘤促生存基因共同参与了AML的发病机制。解剖学研究分子事件及其机制 DNMT3AMut -介导的AML进展应提供关键的洞察新的治疗策略。为了实现这一目标，我们将使用尖端的CRISPR/Cas9技术探讨DNMT3AMut基因增强子DNA低甲基化在诱发基因突变中的作用激活和促进AML决定DNMT3AMut激活的基因通路是什么急性髓系白血病在小鼠体内和体外发育所必需的 (目标1)；我们将通过在我们建立的医院进行功能丧失研究小鼠AML模型(目标2)；第三，在翻译目标中，我们将使用人AML细胞株和原代携带DNMT3AMut的患者来源异种移植模型研究DNMT3AMut对表观基因组的影响人急性髓系白血病的改变、基因表达失控和恶性生长(目标3)。我们希望定义 DNMT3AMut诱导AML表观遗传学/基因改变并有望确定DNMT3AMut的通路促进急性髓系白血病的进展。因为某些已识别的通路，如DOT1L和BCL2，可能可与现有化合物一起下药，我们建议的研究的完成不仅应该促进一个新的机械地理解 DNMT3AMut关联但也将为AML带来创新的治疗方法治疗受影响的癌症患者。

项目成果

期刊论文数量（10）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Epigenetic Perturbations by Arg882-Mutated DNMT3A Potentiate Aberrant Stem Cell Gene-Expression Program and Acute Leukemia Development.

DOI：
10.1016/j.ccell.2016.05.008
发表时间：
2016-07-11
期刊：
Cancer cell
影响因子：
50.3
作者：
Lu R;Wang P;Parton T;Zhou Y;Chrysovergis K;Rockowitz S;Chen WY;Abdel-Wahab O;Wade PA;Zheng D;Wang GG
通讯作者：
Wang GG

R-loop and its functions at the regulatory interfaces between transcription and (epi)genome.

DOI：
10.1016/j.bbagrm.2021.194750
发表时间：
2021-11
期刊：
Biochimica et biophysica acta. Gene regulatory mechanisms
影响因子：
0
作者：
Kim A;Wang GG
通讯作者：
Wang GG

Structural basis for DNMT3A-mediated de novo DNA methylation.

DOI：
10.1038/nature25477
发表时间：
2018-02-15
期刊：
Nature
影响因子：
64.8
作者：
Zhang ZM;Lu R;Wang P;Yu Y;Chen D;Gao L;Liu S;Ji D;Rothbart SB;Wang Y;Wang GG;Song J
通讯作者：
Song J

Polycomb Gene Silencing Mechanisms: PRC2 Chromatin Targeting, H3K27me3 'Readout', and Phase Separation-Based Compaction.