Role of PRC1 in RUNX1-ETO-mediated transcriptional control
PRC1 在 RUNX1-ETO 介导的转录控制中的作用
基本信息
- 批准号:10445091
- 负责人:
- 金额:$ 42.29万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2021
- 资助国家:美国
- 起止时间:2021-07-05 至 2026-06-30
- 项目状态:未结题
- 来源:
- 关键词:AML1-ETO fusion proteinAcute Myelocytic LeukemiaAffectAffinity ChromatographyAllelesBiological AssayCBFA2T1 geneCD34 geneCEBPA geneCRISPR/Cas technologyCancer EtiologyCell Culture TechniquesCell NucleusCellsChimeric ProteinsChromatinChromosomal translocationCloningClustered Regularly Interspaced Short Palindromic RepeatsCollaborationsComplementComplexCoupledCouplingDNA Binding DomainDNA-Directed RNA PolymeraseDRPLA proteinDataDevelopmentDrug CombinationsDrug DesignEnhancersEnvironmentEnzymesEpigenetic ProcessFamily memberFutureGFI1B geneGene ExpressionGenesGeneticGenetic TranscriptionGenomeGenomicsGoalsHDAC3 geneHematopoieticHematopoietic stem cellsHistonesHumanImpairmentIntestinal NeoplasmsKAI1 geneLeukemic CellLinkMass Spectrum AnalysisMediatingMethodsMethylationModelingMolecularMutateMutationMyelogenousNormal CellNuclearNucleosomesOutcomePRC1 ProteinPatientsPhenotypePoint MutationPrognostic MarkerProtacProteinsRNA InterferenceRUNX1 geneReagentRegulator GenesRelapseRepressionRoleRunningSodium ChlorideSolid NeoplasmSystemTestingThalidomideTimeTranscriptTranscriptional RegulationTumor Suppressor ProteinsWorkYeastsanalogbasecell transformationchemical geneticschemotherapydifferential expressionestablished cell linehistone modificationinhibitorinsightleukemogenesismouse geneticsmouse modelnovelprogramsprotein degradationrecruitside effectsmall hairpin RNAsuccesst(821) acute myeloid leukemiat(821)(q22q22)targeted treatmenttooltranscription factortranscriptome sequencingtumorigenesisubiquitin isopeptidaseubiquitin-protein ligaseyeast two hybrid system
项目摘要
The goal of this application is to understand how the t(8;21), the most frequent chromosomal
translocation associated with acute myeloid leukemia (AML), sets the stage for secondary
mutations to accumulate and develop into AML. Understanding how the encoded AML1-ETO
fusion protein alters epigenetic wiring, is critical to finding less debilitating therapies that yield
better outcomes. Both AML1 (RUNX1) and ETO/MTG family members also suffer point mutations
in solid tumors, and the ETO family members Mtgr1 (CBFA2T2) and Mtg16 (CBFA2T3) are tumor
suppressors in mouse models of intestinal neoplasia, so understanding how ETO contacts histone
modifying enzymes has great impact outside of the t(8;21). In our preliminary data, we have used
CRISPR/Cas9 technology to modify the 3’ end of the endogenous AML1-ETO with FKBP12F36V-
HA or 3XFLAG tags to selectively and rapidly degrade AML1-ETO. We have coupled this system
with state-of-the-art genomics such as precision nuclear run-on transcription sequencing (PROseq)
and Cut&Run to establish a chemical genetic system to unambiguously define the mechanism of
transcriptional control by AML1-ETO. Critically, this allows us to define the earliest, and presumably
direct, changes in transcription upon inactivation of the fusion protein. Our preliminary PROseq data
demonstrate that enhancers within key hematopoietic regulatory genes such as CEBPA are
reactivated within 2 hr of adding dTAG47 to Kasumi-1 cell cultures. These novel reagents allow us
to define changes in histone modifications and RNA polymerase dynamics to define the action of
AML1-ETO at defined loci and throughout the genome. Moreover, our preliminary data already
provide a paradigm shift: even though AML1-ETO bound enhancers have been repressed since
the establishment of these cell lines, they were reactivated with a time course that matched the
degradation of the fusion protein. Thus, continued expression of AML1-ETO is needed to maintain
repression, at many loci, while other loci are more permanently silenced. Finally, we used CRISPR
to allow rapid purification of AML1-ETO coupled with MUDPIT and identified a new chromatin
modifying complex as potentially mediating AML1-ETO-dependent repression. We hypothesize that
AML1-ETO recruits histone modifying enzymes to rewire the epigenetic landscape to suppress
CEBPA, PU.1 and GFI1B to impair myeloid differentiation. This sets the stage for secondary
epigenetic mutations that reinforce these changes such as inactivation of ASXL1/2. We will directly
test this hypothesis by defining the molecular contacts that control AML1-ETO recruitment of
repression complexes and use chemical genetics to test if these contacts are required for AML1-
ETO-regulated transcription.
这个应用程序的目标是了解t(8;21),最常见的染色体
与急性髓细胞白血病(AML)相关的易位,为继发性白血病奠定了基础。
突变积累并发展成AML。了解编码的AML 1-ETO
融合蛋白改变了表观遗传连接,对于寻找减少衰弱的治疗方法至关重要,
更好的结果。AML 1(RUNX 1)和ETO/MTG家族成员也存在点突变
在实体瘤中,ETO家族成员Mtgr 1(CBFA 2 T2)和Mtg 16(CBFA 2 T3)是肿瘤
在小鼠肠肿瘤模型中的抑制因子,因此了解ETO如何接触组蛋白
修饰酶在t(8;21)之外具有很大的影响。在我们的初步数据中,
CRISPR/Cas9技术用FKBP 12 F36 V修饰内源性AML 1-ETO的3'末端-
HA或3XFLAG标签选择性和快速降解AML 1-ETO。我们把这个系统
利用最先进的基因组学,如精确核连续转录测序(PROseq)
和Cut&Run建立一个化学遗传系统,以明确定义
AML 1-ETO的转录控制。重要的是,这使我们能够定义最早的,
在融合蛋白失活后直接改变转录。我们的初步PROseq数据
证明了关键造血调控基因(如CEBPA)中的增强子
在向Kasumi-1细胞培养物中加入dTAG 47后2小时内重新活化。这些新试剂让我们
定义组蛋白修饰和RNA聚合酶动力学的变化,以定义
AML 1-ETO在确定的基因座和整个基因组中。此外,我们的初步数据已经
提供了一个范式转变:尽管AML 1-ETO结合增强子自
这些细胞系的建立,它们被重新激活的时间过程,
融合蛋白的降解。因此,需要AML 1-ETO的持续表达来维持
抑制,在许多位点,而其他位点更永久沉默。最后,我们用CRISPR
允许快速纯化与MUDPIT偶联的AML 1-ETO,并鉴定了一种新的染色质
修饰复合物可能介导AML 1-ETO依赖性抑制。我们假设
AML 1-ETO招募组蛋白修饰酶重新连接表观遗传景观,以抑制
CEBPA、PU.1和GFI 1B损害骨髓分化。这就为中学的发展奠定了基础。
表观遗传突变加强了这些变化,如ASXL 1/2的失活。我们会直接
通过定义控制AML 1-ETO募集的分子接触来验证这一假设。
阻遏复合物,并使用化学遗传学来测试这些接触是否需要AML 1-
ETO调节的转录。
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
数据更新时间:{{ journalArticles.updateTime }}
{{
item.title }}
{{ item.translation_title }}
- DOI:
{{ item.doi }} - 发表时间:
{{ item.publish_year }} - 期刊:
- 影响因子:{{ item.factor }}
- 作者:
{{ item.authors }} - 通讯作者:
{{ item.author }}
数据更新时间:{{ journalArticles.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ monograph.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ sciAawards.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ conferencePapers.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ patent.updateTime }}
SCOTT W HIEBERT其他文献
SCOTT W HIEBERT的其他文献
{{
item.title }}
{{ item.translation_title }}
- DOI:
{{ item.doi }} - 发表时间:
{{ item.publish_year }} - 期刊:
- 影响因子:{{ item.factor }}
- 作者:
{{ item.authors }} - 通讯作者:
{{ item.author }}
{{ truncateString('SCOTT W HIEBERT', 18)}}的其他基金
Role of PRC1 in RUNX1-ETO-mediated transcriptional control
PRC1 在 RUNX1-ETO 介导的转录控制中的作用
- 批准号:
10298288 - 财政年份:2021
- 资助金额:
$ 42.29万 - 项目类别:
Role of PRC1 in RUNX1-ETO-mediated transcriptional control
PRC1 在 RUNX1-ETO 介导的转录控制中的作用
- 批准号:
10652591 - 财政年份:2021
- 资助金额:
$ 42.29万 - 项目类别:
Regulation of transcription and tumor suppression by MTG family members
MTG 家族成员对转录和肿瘤抑制的调节
- 批准号:
9265416 - 财政年份:2014
- 资助金额:
$ 42.29万 - 项目类别:
Regulation of transcription and tumor suppression by MTG family members
MTG 家族成员对转录和肿瘤抑制的调节
- 批准号:
8696545 - 财政年份:2014
- 资助金额:
$ 42.29万 - 项目类别:
Regulation of transcription and tumor suppression by MTG family members
MTG 家族成员对转录和肿瘤抑制的调节
- 批准号:
9055662 - 财政年份:2014
- 资助金额:
$ 42.29万 - 项目类别:
FASEB SRC on HDACs, Sirtuins and Reversible Acetylation in Signaling and Disease
FASEB SRC 关于 HDAC、Sirtuins 以及信号传导和疾病中的可逆乙酰化
- 批准号:
8595760 - 财政年份:2013
- 资助金额:
$ 42.29万 - 项目类别:
Hdac3 as a therapeutic target in BCL6-dependent lymphoma
Hdac3 作为 BCL6 依赖性淋巴瘤的治疗靶点
- 批准号:
8607465 - 财政年份:2012
- 资助金额:
$ 42.29万 - 项目类别:
Establishing that Hdac3 is a therapeutic target in a subset of B cell Lymphoma
确定 Hdac3 是 B 细胞淋巴瘤亚型的治疗靶点
- 批准号:
9924498 - 财政年份:2012
- 资助金额:
$ 42.29万 - 项目类别:
Hdac3 as a therapeutic target in BCL6-dependent lymphoma
Hdac3 作为 BCL6 依赖性淋巴瘤的治疗靶点
- 批准号:
8431792 - 财政年份:2012
- 资助金额:
$ 42.29万 - 项目类别:
Hdac3 as a therapeutic target in BCL6-dependent lymphoma
Hdac3 作为 BCL6 依赖性淋巴瘤的治疗靶点
- 批准号:
8230922 - 财政年份:2012
- 资助金额:
$ 42.29万 - 项目类别:
相似海外基金
Computing analysis of leukemic stem cell dynamics in acute myelocytic leukemia
急性粒细胞白血病白血病干细胞动力学的计算分析
- 批准号:
19K08356 - 财政年份:2019
- 资助金额:
$ 42.29万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
Generation of immunotoxins with super-targeting mAb in the acute myelocytic leukemia
在急性髓细胞白血病中使用超靶向单克隆抗体产生免疫毒素
- 批准号:
23501309 - 财政年份:2011
- 资助金额:
$ 42.29万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
DETERMINANTS OF RESPONSE OF ACUTE MYELOCYTIC LEUKEMIA
急性粒细胞白血病反应的决定因素
- 批准号:
3556971 - 财政年份:1980
- 资助金额:
$ 42.29万 - 项目类别:
DETERMINANTS OF RESPONSE OF ACUTE MYELOCYTIC LEUKEMIA
急性粒细胞白血病反应的决定因素
- 批准号:
3556968 - 财政年份:1980
- 资助金额:
$ 42.29万 - 项目类别:
ERADICATION OF ACUTE MYELOCYTIC LEUKEMIA CELLS BY MAB THERAPY
通过 MAB 疗法根除急性粒细胞白血病细胞
- 批准号:
3889304 - 财政年份:
- 资助金额:
$ 42.29万 - 项目类别: