Mechanisms of HbF Activation by Non-deletional HPFH
非缺失 HPFH 激活 HbF 的机制
基本信息
- 批准号:8721479
- 负责人:
- 金额:$ 37万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2013
- 资助国家:美国
- 起止时间:2013-08-15 至 2017-05-31
- 项目状态:已结题
- 来源:
- 关键词:3-DimensionalAdultAdverse effectsAffectAfrican AmericanAmino AcidsBindingBinding SitesBiochemicalBiological AssayBone Marrow CellsCell Culture TechniquesCellsChromatinChromatin StructureChromosomes, Artificial, YeastClinicalComplexCoupledDNA BindingDNA-Binding ProteinsDataDeoxyribonuclease IDevelopmentDiseaseDistalEMSAEnvironmentErythropoiesisEthylnitrosoureaFetal HemoglobinFluorescence-Activated Cell SortingFunctional disorderGene ActivationGene ExpressionGene SilencingGenesGlobinGoalsGreekGreen Fluorescent ProteinsGrowthHemoglobin F DiseaseHereditary DiseaseHumanHypersensitivityIntercistronic RegionKnowledgeLabelLeadLightLocus Control RegionMass Spectrum AnalysisModelingModificationMolecularMolecular BiologyMolecular ConformationMolecular TargetMusMutagenesisMutationMutation SpectraNatureNucleotidesOutcome StudyPatientsPhenotypePoint MutationPopulationProcessPromoter RegionsProtein BindingProteinsProteomicsRegulatory ElementReporterReportingRepressionResearchResolutionSickle Cell AnemiaSiteStable Isotope LabelingStagingStem cellsStructureSwitch GenesTestingTherapeuticTranscription Repressor/CorepressorTransgenesTransgenic MiceUp-Regulationbaseeffective therapyembryo/fetusfetalgene interactiongene repressionhematopoietic tissuehistone modificationin vivomeltingmutantnew therapeutic targetnovelpreventpromoterprotein complexpublic health relevancetransgene expression
项目摘要
DESCRIPTION (provided by applicant): Understanding the molecular mechanisms underlying the human ?- to ¿-globin gene switch has long been recognized as important in the treatment of sickle cell disease (SCD), since a wealth of evidence has demonstrated that increased fetal hemoglobin (HbF) significantly decreases the pathophysiology associated with this disease. Thus, knowledge of how to reactivate ?-globin (HbF) in adult erythropoiesis will benefit SCD patients. Our goal in this study is to understand ?-globin gene silencing during the adult stage of
definitive erythropoiesis. Our clinical goal is to identify new molecular targets based on the outcome of this study that can be modulated therapeutically for up-regulation of ?-globin synthesis to treat SCD. Non-deletional hereditary persistence of fetal hemoglobin (HPFH) point mutations are likely to be highly informative regarding mechanisms of ?-globin gene repression and activation, but to date have not been studied extensively at the mechanistic level. The proposed study will test the hypothesis that HPFH mutations prevent silencing or maintain activation of ?-globin gene expression by abrogating recruitment of transcriptional repressor complex ("repressosome") components normally located at the ?-globin gene or more distal intergenic regions of the locus, or alternately, by creating a favorable chromatin structure that allows the ?-globin genes to partly outcompete the ¿-globin gene for interaction with the locus control region (LCR), or both. Several HPFH mutations have been introduced into our human ¿-globin locus yeast artificial chromosome (¿-YAC) including the -566, -195, -175, and -117 ,?-globin non-deletional HPFH point mutations and transgenic mice have been produced. In Specific Aim 1, we will study the mechanisms by which the - 566, -195, -175, and -117 HPFH mutations disrupt ?-globin gene repression and/or alter the ?-globin locus chromatin domain during development in vivo using murine models and molecular biology/biochemical approaches including transgene structure/expression studies, 3C, histone modification, DNase I sensitivity, co-activator/repressor recruitment, Bcl11A recruitment, and synergy between the HPFH mutations. In Specific Aim 2 we will examine how these four HPFH mutations alter DNA-binding protein complexes in the A?-globin promoter using proteomics of isolated chromatin segments (PICh) and Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC) coupled with mass spectrometry (MS). In Specific Aim 3, we will generate and identify novel HPFH mutations within the A?-globin gene promoter using a cell-based reporter assay to select for HPFH mutations, followed by phenotypic characterization of these mutations in ¿-YAC transgenic mice and determination of the DNA-binding activity at these sites. The knowledge we gain from these studies will reveal novel therapeutic targets for which highly-specific treatments may be developed to increase HbF for the treatment of SCD without the side-effects associated with broad-spectrum therapies.
描述(由申请人提供):了解人类?至β-珠蛋白基因开关长期以来被认为在镰状细胞病(SCD)的治疗中是重要的,因为大量证据已经证明增加的胎儿血红蛋白(HbF)显著降低与这种疾病相关的病理生理学。所以,知道如何重新激活?成人红细胞生成中的血红蛋白(HbF)将有益于SCD患者。我们在这项研究中的目标是了解?珠蛋白基因沉默在成年期
确定性红细胞生成我们的临床目标是根据这项研究的结果确定新的分子靶点,这些靶点可以在治疗上进行调节,以上调?珠蛋白合成来治疗SCD。胎儿血红蛋白(HPFH)点突变的非缺失性遗传持续性可能是关于?珠蛋白基因的抑制和激活,但迄今尚未在机制水平上进行广泛研究。这项拟议的研究将检验HPFH突变阻止沉默或维持激活的假设。球蛋白基因的表达,通过取消募集转录阻遏复合物(“阻遏体”)的组成部分,通常位于?珠蛋白基因或基因座的更远端的基因间区域,或者,通过创建有利的染色质结构,珠蛋白基因与基因座控制区(LCR)的相互作用部分地胜过β-珠蛋白基因,或两者兼而有之。几个HPFH突变已被引入到我们的人类<$-珠蛋白基因座酵母人工染色体(<$-YAC)中,包括-566,-195,-175和-117,?已经产生了珠蛋白非缺失HPFH点突变和转基因小鼠。在具体目标1中,我们将研究-566、-195、-175和-117 HPFH突变破坏?珠蛋白基因抑制和/或改变?使用小鼠模型和分子生物学/生物化学方法,包括转基因结构/表达研究、3C、组蛋白修饰、DNA酶I敏感性、共激活子/阻遏物募集、Bcl 11 A募集和HPFH突变之间的协同作用,在体内发育过程中对珠蛋白基因座染色质结构域的影响。在具体目标2中,我们将研究这四个HPFH突变如何改变A?使用分离的染色质片段的蛋白质组学(PICh)和细胞培养物中氨基酸的稳定同位素标记(SILAC)结合质谱(MS)对珠蛋白启动子进行分析。在具体目标3中,我们将在A?珠蛋白基因启动子使用基于细胞的报告基因测定来选择HPFH突变,随后在<$-YAC转基因小鼠中对这些突变进行表型表征并测定这些位点处的DNA结合活性。我们从这些研究中获得的知识将揭示新的治疗靶点,可以开发高度特异性的治疗方法,以增加HbF治疗SCD,而不会产生与广谱治疗相关的副作用。
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 }}
KENNETH R PETERSON其他文献
KENNETH R PETERSON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('KENNETH R PETERSON', 18)}}的其他基金
Regulation of Globin Gene Switching by O-GlcNAc Post-Translational Modification
O-GlcNAc 翻译后修饰调控球蛋白基因转换
- 批准号:
8610686 - 财政年份:2014
- 资助金额:
$ 37万 - 项目类别:
Regulation of Globin Gene Switching by O-GlcNAc Post-Translational Modification
O-GlcNAc 翻译后修饰调控球蛋白基因转换
- 批准号:
8784216 - 财政年份:2014
- 资助金额:
$ 37万 - 项目类别:
Regulation of Globin Gene Switching by O-GlcNAc Post-Translational Modification
O-GlcNAc 翻译后修饰调控球蛋白基因转换
- 批准号:
8995201 - 财政年份:2014
- 资助金额:
$ 37万 - 项目类别:
Mechanisms of HbF Activation by Non-deletional HPFH
非缺失 HPFH 激活 HbF 的机制
- 批准号:
8578291 - 财政年份:2013
- 资助金额:
$ 37万 - 项目类别:
Mechanisms of HbF Activation by Non-deletional HPFH
非缺失 HPFH 激活 HbF 的机制
- 批准号:
8854128 - 财政年份:2013
- 资助金额:
$ 37万 - 项目类别:
相似海外基金
Co-designing a lifestyle, stop-vaping intervention for ex-smoking, adult vapers (CLOVER study)
为戒烟的成年电子烟使用者共同设计生活方式、戒烟干预措施(CLOVER 研究)
- 批准号:
MR/Z503605/1 - 财政年份:2024
- 资助金额:
$ 37万 - 项目类别:
Research Grant
Early Life Antecedents Predicting Adult Daily Affective Reactivity to Stress
早期生活经历预测成人对压力的日常情感反应
- 批准号:
2336167 - 财政年份:2024
- 资助金额:
$ 37万 - 项目类别:
Standard Grant
RAPID: Affective Mechanisms of Adjustment in Diverse Emerging Adult Student Communities Before, During, and Beyond the COVID-19 Pandemic
RAPID:COVID-19 大流行之前、期间和之后不同新兴成人学生社区的情感调整机制
- 批准号:
2402691 - 财政年份:2024
- 资助金额:
$ 37万 - 项目类别:
Standard Grant
Elucidation of Adult Newt Cells Regulating the ZRS enhancer during Limb Regeneration
阐明成体蝾螈细胞在肢体再生过程中调节 ZRS 增强子
- 批准号:
24K12150 - 财政年份:2024
- 资助金额:
$ 37万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
Migrant Youth and the Sociolegal Construction of Child and Adult Categories
流动青年与儿童和成人类别的社会法律建构
- 批准号:
2341428 - 财政年份:2024
- 资助金额:
$ 37万 - 项目类别:
Standard Grant
Understanding how platelets mediate new neuron formation in the adult brain
了解血小板如何介导成人大脑中新神经元的形成
- 批准号:
DE240100561 - 财政年份:2024
- 资助金额:
$ 37万 - 项目类别:
Discovery Early Career Researcher Award
Laboratory testing and development of a new adult ankle splint
新型成人踝关节夹板的实验室测试和开发
- 批准号:
10065645 - 财政年份:2023
- 资助金额:
$ 37万 - 项目类别:
Collaborative R&D
Usefulness of a question prompt sheet for onco-fertility in adolescent and young adult patients under 25 years old.
问题提示表对于 25 岁以下青少年和年轻成年患者的肿瘤生育力的有用性。
- 批准号:
23K09542 - 财政年份:2023
- 资助金额:
$ 37万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
Identification of new specific molecules associated with right ventricular dysfunction in adult patients with congenital heart disease
鉴定与成年先天性心脏病患者右心室功能障碍相关的新特异性分子
- 批准号:
23K07552 - 财政年份:2023
- 资助金额:
$ 37万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
Issue identifications and model developments in transitional care for patients with adult congenital heart disease.
成人先天性心脏病患者过渡护理的问题识别和模型开发。
- 批准号:
23K07559 - 财政年份:2023
- 资助金额:
$ 37万 - 项目类别:
Grant-in-Aid for Scientific Research (C)