NUCLEAR CAPTURE FOR CELL-TYPE SPECIFIC EPIGENETIC ANALYSIS OF ADIPOCYTES
用于脂肪细胞细胞类型特异性表观遗传分析的核捕获
基本信息
- 批准号:8917210
- 负责人:
- 金额:$ 31.41万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2014
- 资助国家:美国
- 起止时间:2014-08-26 至 2018-06-30
- 项目状态:已结题
- 来源:
- 关键词:AddressAdipocytesAdipose tissueAdultAffectAffinityAntibodiesAntibody AffinityAntigensBloodBone MarrowBrown FatCandidate Disease GeneCell LineCell NucleusCell SeparationCellsChromatinChromatin StructureCognition DisordersCompanionsCytometryDNA MethylationDataDevelopmentDiabetes MellitusDiagnosticDiagnostics ResearchDiseaseDisease ProgressionEndocrine GlandsEnvironmentEpigenetic ProcessEpitopesFluorescenceGene ExpressionGene Expression ProfileGenesGeneticGenetic TranscriptionGoalsGrantHealthHumanHuman GenomeHybridomasIndividualInflammationInvestigationMalignant NeoplasmsMedical ResearchMedicineMembraneMembrane ProteinsMetabolicMetabolic syndromeMethodologyMethodsMicroscopyModificationMolecularMolecular GeneticsMolecular ProfilingMonoclonal AntibodiesMorphologyMusMuscleMutationNeurobiologyNeurodegenerative DisordersNeuronsNuclearNuclear EnvelopeObesityPlayProductionPropertyProteomeReagentRecoveryResearch Project GrantsRoleSamplingSorting - Cell MovementSpecificityStagingT-LymphocyteTechnologyTestingTherapeuticTimeTissuesVisceralWorkadipokinesadiponectinbasecardiovascular disorder riskcell typecostdisorder riskepigenomegenome-wideinsulin sensitivitymammalian genomemeetingsmethylometechnology developmenttherapeutic development
项目摘要
DESCRIPTION (provided by applicant): This proposal addresses two important issues: the technical issue of isolating specific types of adipocytes from adipose depots and other tissues (e.g., muscle, bone marrow) for genetic and epigenetic analysis, and the issue of determining epigenetic changes to subclasses of adipocytes that are associated with obesity. Epigenetic reprogramming of adipocytes, particularly in visceral adipose depots, plays an important role in the transformation of adipose tissue to a metabolically dysfunctional state contributing to increased systemic inflammation and reduced insulin sensitivity. However, the epigenetic reprogramming of subclasses of adipocytes is essentially unexplored due to the technical difficulty of isolating these cells. We are proposing a powerful technology, Capture by Nuclear Antibody (CANA) that will enable cell type-specific epigenetic analysis of different classes of adipocytes. CANA is based on the fact that mammalian genomes encode more than two thousand nuclear trans-membrane proteins, that cell types differ in the expression of these "nuclear membrane antigens (NMAs)", and NMAs may serve as nuclear immuno-epitope tags for the purification of specific nuclei. We hypothesize that subclasses of adipocytes are epigenetically reprogrammed in obese individuals. The goals of this technology development grant are to (1) generate a battery of adipocyte nuclear membrane antigen-specific monoclonal antibodies (mAbs) to affinity purify distinct classes of human adipocyte nuclei and (2) investigate
differences in DNA methylation and RNA expression profiles within two subclasses of human visceral adipocyte nuclei that are associated with obesity. We will meet these goals and set the stage for further testing our hypothesis by pursuing the following Specific Aims: Aim 1. Develop CANA reagent mAbs with clear nuclear subclass specificities. DiSH" (Direct Selection of Hybridomas) technology, which enables the efficient and rapid production of hundreds of mAbs to membrane antigens, will provide more than 200 anti-nuclear (anti-NU) mAbs to nuclei from human visceral adipose tissue and mouse beige and brown adipocyte cell lines. Those with the most useful nuclear subtype specificities will be selected and these mAbs will be used to affinity-purify two subclasses of human visceral adipocyte nuclei. Aim 2. Characterize the genome-wide DNA methylation and RNA expression profiles in two specific subclasses of nuclei affinity purified from the visceral adipose tissue of obese and lean individuals. Epigenetic and gene expression data will be integrated to identify obesity-associated genes. CANA technology addresses one of the greatest challenges to neurobiology today, the difficulty of working with subsets of adult neurons. It will contribute significantly to medical research, diagnostic and therapeutic development in studies of numerous cognitive disorders, including obesity. Once established, the methodology can be expanded to support molecular studies of specific cell types throughout the body.
描述(由申请人提供):该提案解决了两个重要问题:从脂肪库和其他组织中分离特定类型脂肪细胞的技术问题(例如,肌肉,骨髓)进行遗传和表观遗传分析,以及确定与肥胖相关的脂肪细胞亚类的表观遗传变化的问题。脂肪细胞的表观遗传重编程,特别是在内脏脂肪库中,在脂肪组织转化为代谢功能障碍状态中起重要作用,从而导致全身炎症增加和胰岛素敏感性降低。然而,由于分离这些细胞的技术困难,脂肪细胞亚类的表观遗传重编程基本上未被探索。我们正在提出一种强大的技术,即核抗体捕获(CANA),它将使不同类型脂肪细胞的细胞类型特异性表观遗传分析成为可能。CANA基于哺乳动物基因组编码超过2000种核跨膜蛋白,细胞类型在这些“核膜抗原(NMA)"的表达方面不同,并且NMA可以用作用于纯化特定核的核免疫表位标签的事实。我们推测,脂肪细胞的亚类在肥胖个体中表观遗传学上被重编程。这项技术开发资助的目标是(1)产生一组脂肪细胞核膜抗原特异性单克隆抗体(mAb),以亲和纯化不同类别的人类脂肪细胞核,以及(2)研究
在与肥胖相关的人类内脏脂肪细胞核的两个亚类内的DNA甲基化和RNA表达谱的差异。我们将通过追求以下具体目标来实现这些目标,并为进一步检验我们的假设奠定基础:目标1。开发具有明确核亚类特异性的CANA试剂mAb。DiSH”(杂交瘤直接选择)技术能够高效快速地生产数百种针对膜抗原的mAb,将为人类内脏脂肪组织和小鼠米色和棕色脂肪细胞系的细胞核提供200多种抗核(抗NU)mAb。将选择具有最有用的核亚型特异性的那些,并且这些mAb将用于亲和纯化人内脏脂肪细胞核的两个亚类。目标2.描述从肥胖和瘦个体的内脏脂肪组织中纯化的两个特定核亲和力亚类的全基因组DNA甲基化和RNA表达谱。表观遗传学和基因表达数据将被整合,以确定肥胖相关基因。CANA技术解决了当今神经生物学面临的最大挑战之一,即处理成年神经元子集的困难。它将为包括肥胖症在内的许多认知障碍的医学研究、诊断和治疗发展做出重大贡献。一旦建立,该方法可以扩展到支持整个身体的特定细胞类型的分子研究。
项目成果
期刊论文数量(1)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
The Genome and Methylome of a Beetle with Complex Social Behavior, Nicrophorus vespilloides (Coleoptera: Silphidae).
- DOI:10.1093/gbe/evv194
- 发表时间:2015-10-09
- 期刊:
- 影响因子:3.3
- 作者:Cunningham CB;Ji L;Wiberg RA;Shelton J;McKinney EC;Parker DJ;Meagher RB;Benowitz KM;Roy-Zokan EM;Ritchie MG;Brown SJ;Schmitz RJ;Moore AJ
- 通讯作者:Moore AJ
{{
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 }}
Richard Brian Meagher其他文献
Richard Brian Meagher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('Richard Brian Meagher', 18)}}的其他基金
Targeted pan-antibacterial liposomes to control pathogenic mycobacteria
靶向泛抗菌脂质体控制致病性分枝杆菌
- 批准号:
10569282 - 财政年份:2023
- 资助金额:
$ 31.41万 - 项目类别:
Targeted delivery of antifungal drug loaded liposomes to control mucormycosis
靶向递送抗真菌药物脂质体来控制毛霉菌病
- 批准号:
10668509 - 财政年份:2022
- 资助金额:
$ 31.41万 - 项目类别:
Targeted delivery of antifungal drug loaded liposomes to control mucormycosis
靶向递送抗真菌药物脂质体来控制毛霉菌病
- 批准号:
10509943 - 财政年份:2022
- 资助金额:
$ 31.41万 - 项目类别:
Antifungal Liposomes Targeted to Cell Wall Mannans
针对细胞壁甘露聚糖的抗真菌脂质体
- 批准号:
9979038 - 财政年份:2020
- 资助金额:
$ 31.41万 - 项目类别:
NUCLEAR CAPTURE FOR CELL-TYPE SPECIFIC EPIGENETIC ANALYSIS OF ADIPOCYTES
用于脂肪细胞细胞类型特异性表观遗传分析的核捕获
- 批准号:
8760476 - 财政年份:2014
- 资助金额:
$ 31.41万 - 项目类别:
Differential Expression of the Diverse Plant Actins
多种植物肌动蛋白的差异表达
- 批准号:
7931495 - 财政年份:2009
- 资助金额:
$ 31.41万 - 项目类别:
DIFFERENTIAL EXPRESSION OF DIVERSE PLANT ACTIN GENES
多种植物肌动蛋白基因的差异表达
- 批准号:
6039284 - 财政年份:1986
- 资助金额:
$ 31.41万 - 项目类别:
DIFFERENTIAL EXPRESSION OF DIVERSE PLANT ACTIN GENES
多种植物肌动蛋白基因的差异表达
- 批准号:
2684820 - 财政年份:1986
- 资助金额:
$ 31.41万 - 项目类别:
相似国自然基金
支链氨基酸代谢紊乱调控“Adipocytes - Macrophages Crosstalk”诱发2型糖尿病脂肪组织功能和结构障碍的作用及机制
- 批准号:81970721
- 批准年份:2019
- 资助金额:55.0 万元
- 项目类别:面上项目
相似海外基金
Recruitment of brown adipocytes in visceral white adipose tissue by fibroblast growth factor 8b
成纤维细胞生长因子 8b 将棕色脂肪细胞募集到内脏白色脂肪组织中
- 批准号:
321208980 - 财政年份:2016
- 资助金额:
$ 31.41万 - 项目类别:
Research Grants
Enhancing Energy Expending Adipocytes in White Adipose Tissue
增强白色脂肪组织中的能量消耗脂肪细胞
- 批准号:
8827438 - 财政年份:2014
- 资助金额:
$ 31.41万 - 项目类别:
Induction of brown-like adipocytes in white adipose tissue by food-derived factors
食物源性因子在白色脂肪组织中诱导棕色样脂肪细胞
- 批准号:
26450168 - 财政年份:2014
- 资助金额:
$ 31.41万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
WAT-on-a-chip - Development of a micofluidic, microphysiologic in vitro adipose tissue model for high-throughput drug screening based on hiPSC-derived adipocytes.
WAT-on-a-chip - 开发微流体、微生理体外脂肪组织模型,用于基于 hiPSC 衍生脂肪细胞的高通量药物筛选。
- 批准号:
257256526 - 财政年份:2014
- 资助金额:
$ 31.41万 - 项目类别:
Research Fellowships
Enhancing Energy Expending Adipocytes in White Adipose Tissue
增强白色脂肪组织中的能量消耗脂肪细胞
- 批准号:
8828181 - 财政年份:2013
- 资助金额:
$ 31.41万 - 项目类别:
Enhancing Energy Expending Adipocytes in White Adipose Tissue
增强白色脂肪组织中的能量消耗脂肪细胞
- 批准号:
8520690 - 财政年份:2013
- 资助金额:
$ 31.41万 - 项目类别:
Enhancing Energy Expending Adipocytes in White Adipose Tissue
增强白色脂肪组织中的能量消耗脂肪细胞
- 批准号:
8629741 - 财政年份:2013
- 资助金额:
$ 31.41万 - 项目类别:
Effect of exercise training on formation of brite adipocytes within white adipose tissue
运动训练对白色脂肪组织内脂肪细胞形成的影响
- 批准号:
23700778 - 财政年份:2011
- 资助金额:
$ 31.41万 - 项目类别:
Grant-in-Aid for Young Scientists (B)
Investigation for the mechanisms of the emergence of brown adipocytes in white adipose tissue
白色脂肪组织中棕色脂肪细胞出现机制的研究
- 批准号:
21780261 - 财政年份:2009
- 资助金额:
$ 31.41万 - 项目类别:
Grant-in-Aid for Young Scientists (B)
LOUISIANA COBRE: P1: INDUCE THERMOGENIC BROWN ADIPOCYTES IN WHITE ADIPOSE TISSUE
路易斯安那 COBRE:P1:在白色脂肪组织中诱导产热棕色脂肪细胞
- 批准号:
7610781 - 财政年份:2007
- 资助金额:
$ 31.41万 - 项目类别: