Dissecting the molecular mechanism of heterochromatin misregulation at the nuclear periphery in cancer
剖析癌症核周边异染色质失调的分子机制
基本信息
- 批准号:10021632
- 负责人:
- 金额:$ 4.06万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2019
- 资助国家:美国
- 起止时间:2019-09-20 至 2020-12-31
- 项目状态:已结题
- 来源:
- 关键词:ArchitectureAtomic Force MicroscopyBehaviorBindingBiochemicalBiochemistryBiological AssayBiological ModelsBiologyBiophysical ProcessBiophysicsBreast Cancer PatientBreast Cancer cell lineBreast cancer metastasisCRISPR interferenceCancer cell lineCell LineCell NucleusCell physiologyCellsCharacteristicsChromatinChromatin FiberChromatin StructureChromosomesComplexConfocal MicroscopyCoupledDNADNA-Protein InteractionDevelopmentDiseaseDistalDown-RegulationElectron MicroscopyEstrogen receptor positiveExhibitsFluorescence MicroscopyGene Expression RegulationGene SilencingGenesGenetic TranscriptionGenomeGoalsHeterochromatinHistologicHistone H3In VitroIndividualInjectionsIntegral Membrane ProteinIntermediate Filament ProteinsLabelLamin Type BLaminsLengthLiquid substanceLysineMCF7 cellMalignant NeoplasmsMechanicsMediatingMediator of activation proteinMembraneMethylationModificationMolecularMonitorMorphologyMusNeoplasm MetastasisNuclearNuclear EnvelopeNuclear Inner MembraneNuclear PleomorphismNucleosomesOrganismOutputPatient-Focused OutcomesPhasePhysiologicalPlayPleomorphismProtein RegionProteinsRecurrenceResearchResearch Project GrantsResolutionRoleSeveritiesShapesSumSystemT47DTechniquesTertiary Protein StructureTestingTimeTumor MarkersUp-RegulationVariantWorkbiophysical propertiescancer celldiagnostic biomarkerdisease phenotypeexperimental studyheterochromatin-specific nonhistone chromosomal protein HP-1histone modificationin vivolamin B receptormalignant breast neoplasmmigrationmouse modelneoplastic cellnoveloverexpressionphysical modelprogramsprotein expressionpublic health relevancereceptor bindingreconstitutionrecruitrestorationsingle moleculetargeted cancer therapytherapy developmenttumor progression
项目摘要
Project Summary/Abstract
A key driver of cancer is the global deregulation of transcription networks in the cell. Transcriptional output
is dependent on chromatin structure and topology; repressed genes are sequestered from active genes into
regions of compacted DNA near the nuclear membrane. Dense chromosomal domains, termed heterochromatin,
are associated with Heterochromatin Protein-1 (HP1) and are tethered to the periphery via an interaction with
the nuclear transmembrane protein, Lamin Binding Receptor (LBR), and the intermediate filament protein, Lamin
B. Consequently, cancer cells that undergo global rewiring of transcription also often exhibit aberrant protein
expression of Lamin B, LBR and HP1. The physical manifestation of the misregulation of these proteins is
distortion in the size and shape of the nuclear envelope. Indeed, this nuclear pleomorphism is used as a
histological marker of tumor progression. In estrogen receptor positive (ER+) breast cancer patients,
downregulation of HP1 and upregulation of Lamin B and LBR is strongly correlated with earlier occurrence of
distal metastasis. Understanding the biophysical properties that govern the assembly of heterochromatin at the
nuclear periphery will facilitate the development of therapies aimed at restoring proper gene regulation. HP1 was
recently found to concentrate DNA and chromatin into liquid-liquid phase separated (LLPS) droplets in vitro. This
suggests a potential mechanism of DNA organization in vivo. To investigate the molecular details of HP1-
mediated compaction and phase separation, I utilized DNA curtains and confocal microscopy. I identified key
regions of HP1 required for multivalent—LLPS interactions and DNA compaction. This preliminary research
focused on DNA, and the graduate work in Aim 1 will build on these studies by evaluating HP1 interactions with
complex nucleosomal substrates. Heterochromatin in vivo is distinguished by evenly spaced nucleosomes and
the trimethylation of histone H3 lysine 9 (H3K9me3). I will make chromatin substrates that range from mono-
nucleosomes to 50kb chromatin fibers with variations in spacing and methylation modification. I will monitor the
binding, oligomerization and phase separation of HP1 on these substrates by a combination of bulk biochemical
assays and a novel single molecule chromatin assay. The proposed postdoctoral research in Aim 2 will focus
on determining a physical model of the nuclear periphery. I will reconstitute the interactions between chromatin
and the lamina with single molecule studies in vitro and super resolution studies in cells. I will use cell lines and
mouse models of breast cancer metastasis to determine the molecular mechanism guiding metastatic recurrence
of ER+ breast cancer patients with high expression of Lamin B and LBR. This research program will propel me
toward my ultimate goal of leading my own lab studying how nuclear topology is coupled to cell fate determination
in development, and the misregulation that leads to disease.
项目概要/摘要
癌症的一个关键驱动因素是细胞内转录网络的整体失调。转录输出
取决于染色质结构和拓扑;被抑制的基因被从活跃的基因中隔离出来
核膜附近的压缩 DNA 区域。致密染色体结构域,称为异染色质,
与异染色质蛋白 1 (HP1) 相关,并通过与
核跨膜蛋白、核纤层蛋白结合受体 (LBR) 和中间丝蛋白、核纤层蛋白
B. 因此,经历转录全局重连的癌细胞也经常表现出异常蛋白
Lamin B、LBR 和 HP1 的表达。这些蛋白质失调的物理表现是
核膜的大小和形状发生扭曲。事实上,这种核多形性被用作
肿瘤进展的组织学标志物。在雌激素受体阳性(ER+)乳腺癌患者中,
HP1 的下调以及 Lamin B 和 LBR 的上调与早期发生密切相关
远端转移。了解控制异染色质组装的生物物理特性
核外围将促进旨在恢复适当基因调控的疗法的开发。 HP1 是
最近发现可以在体外将 DNA 和染色质浓缩到液-液相分离 (LLPS) 液滴中。这
提出了体内 DNA 组织的潜在机制。研究 HP1- 的分子细节
为了介导压实和相分离,我利用了 DNA 帘和共焦显微镜。我确定了关键
多价-LLPS 相互作用和 DNA 压缩所需的 HP1 区域。本次初步研究
重点关注 DNA,目标 1 的研究生工作将通过评估 HP1 与
复杂的核小体底物。体内异染色质的特征是均匀分布的核小体和
组蛋白 H3 赖氨酸 9 (H3K9me3) 的三甲基化。我将制作从单染色质底物到
核小体到 50kb 染色质纤维,间距和甲基化修饰各不相同。我会监控
通过大量生化的组合,HP1 在这些底物上的结合、寡聚化和相分离
测定和新型单分子染色质测定。目标 2 中拟议的博士后研究将集中于
确定核外围的物理模型。我将重建染色质之间的相互作用
以及在体外进行单分子研究和在细胞中进行超分辨率研究的层板。我将使用细胞系和
乳腺癌转移小鼠模型以确定指导转移复发的分子机制
Lamin B 和 LBR 高表达的 ER+ 乳腺癌患者。这个研究计划将推动我
我的最终目标是领导自己的实验室研究核拓扑结构如何与细胞命运决定相结合
发育过程中的失调,以及导致疾病的失调。
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 }}
Madeline Keenen其他文献
Madeline Keenen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('Madeline Keenen', 18)}}的其他基金
Dissecting the mechanistic role of multinucleation in breast and trophoblast cancers
剖析多核在乳腺癌和滋养层细胞癌中的机制作用
- 批准号:
10321010 - 财政年份:2021
- 资助金额:
$ 4.06万 - 项目类别:
Dissecting the mechanistic role of multinucleation in breast and trophoblast cancers
剖析多核在乳腺癌和滋养层细胞癌中的机制作用
- 批准号:
10549720 - 财政年份:2021
- 资助金额:
$ 4.06万 - 项目类别:
Dissecting the mechanistic role of multinucleation in breast and trophoblast cancers
剖析多核在乳腺癌和滋养层细胞癌中的机制作用
- 批准号:
10295856 - 财政年份:2021
- 资助金额:
$ 4.06万 - 项目类别:
Dissecting the mechanistic role of multinucleation in breast and trophoblast cancers
剖析多核在乳腺癌和滋养层细胞癌中的机制作用
- 批准号:
10834418 - 财政年份:2021
- 资助金额:
$ 4.06万 - 项目类别:
相似海外基金
Pushing the envelope: atomic force microscopy imaging of the bacterial outer membrane during growth and division
挑战极限:生长和分裂过程中细菌外膜的原子力显微镜成像
- 批准号:
BB/X007669/1 - 财政年份:2024
- 资助金额:
$ 4.06万 - 项目类别:
Research Grant
Nanoscopic elucidation of dynamic behavior of RNA viral nucleocapsid proteins using high-speed atomic force microscopy (HS-AFM)
使用高速原子力显微镜 (HS-AFM) 纳米级阐明 RNA 病毒核衣壳蛋白的动态行为
- 批准号:
24K18449 - 财政年份:2024
- 资助金额:
$ 4.06万 - 项目类别:
Grant-in-Aid for Early-Career Scientists
Unravelling dengue virus structural dynamics and conformational changes using high-speed atomic force microscopy
使用高速原子力显微镜揭示登革热病毒结构动力学和构象变化
- 批准号:
24K18450 - 财政年份:2024
- 资助金额:
$ 4.06万 - 项目类别:
Grant-in-Aid for Early-Career Scientists
Atomic scale reactivity of small islands of a bimetallic alloy on ceria to small molecules investigated by ultrahigh resolution atomic force microscopy
通过超高分辨率原子力显微镜研究二氧化铈上双金属合金小岛对小分子的原子尺度反应性
- 批准号:
24K01350 - 财政年份:2024
- 资助金额:
$ 4.06万 - 项目类别:
Grant-in-Aid for Scientific Research (B)
State-of-the-art atomic force microscopy facilities for South Australia
南澳大利亚最先进的原子力显微镜设施
- 批准号:
LE240100129 - 财政年份:2024
- 资助金额:
$ 4.06万 - 项目类别:
Linkage Infrastructure, Equipment and Facilities
Pushing the envelope: atomic force microscopy imaging of the bacterial outer membrane during growth and division
挑战极限:生长和分裂过程中细菌外膜的原子力显微镜成像
- 批准号:
BB/X00760X/1 - 财政年份:2024
- 资助金额:
$ 4.06万 - 项目类别:
Research Grant
A New Nano Tip Fabrication Technique for Atomic Force Microscopy
原子力显微镜的新型纳米尖端制造技术
- 批准号:
DP230100637 - 财政年份:2023
- 资助金额:
$ 4.06万 - 项目类别:
Discovery Projects
Magnetic imaging by the locally induced anomalous Nernst effect using atomic force microscopy
使用原子力显微镜通过局部诱发的异常能斯特效应进行磁成像
- 批准号:
23K04579 - 财政年份:2023
- 资助金额:
$ 4.06万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
Characterization of super adhesive aerosols on the basis of individual particle analysis using atomic force microscopy
基于原子力显微镜单个颗粒分析的超粘性气溶胶表征
- 批准号:
22KJ1464 - 财政年份:2023
- 资助金额:
$ 4.06万 - 项目类别:
Grant-in-Aid for JSPS Fellows
Using atomic force microscopy to explore the processes and re-organisations that occur during bacterial growth and division and how these are influenc
使用原子力显微镜探索细菌生长和分裂过程中发生的过程和重组以及它们如何影响细菌
- 批准号:
2887441 - 财政年份:2023
- 资助金额:
$ 4.06万 - 项目类别:
Studentship