Histone deacetylase 7 and its interacting proteins in endothelial cells

内皮细胞中组蛋白脱乙酰酶 7 及其相互作用蛋白

基本信息

批准号：
7780590
负责人：
HUNG-YING KAO
金额：
$ 39.25万
依托单位：
CASE WESTERN RESERVE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-02-01 至 2014-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7780590
关键词：
Acute Promyelocytic Leukemia Adhesions Affect Animal Model Apoptosis Apoptotic Binding Biological Biological Assay Blood Vessels Cell Nucleus Cell Proliferation Chemicals Complex Cytokine Activation Cytokine Receptors Cytoplasm DNA Sequence Development Dilatation - action Dominant-Negative Mutation Drug or chemical Tissue Distribution Embryo Endothelial Cells Fibroblasts Gene Expression Gene Targeting Genetic Transcription Goals HDAC4 gene Histone Deacetylase Histones Human In Vitro Infection Inflammation Inflammatory Injury Interferons Interleukins Ionizing radiation Knock-out Knockout Mice Link Lipopolysaccharides Mediating Mediator of activation protein Messenger RNA Modeling Monocyte Chemoattractant Protein-1 Mus Muscle Cells Nuclear Oxidative Stress Pathway interactions Play Proteins Regulation Reporter Repression Role Rupture Series Signal Pathway Signal Transduction Site Skin Carcinogenesis Small Interfering RNA Stimulus Testing Therapeutic Transcriptional Regulation Transfection Tube Tumor Necrosis Factor-alpha Vascular Diseases Vascular Endothelial Cell Virus angiogenesis cell growth cell motility chromatin immunoprecipitation cytokine derepression extracellular histone modification human disease in vivo inhibitor/antagonist interest irradiation member migration monocyte novel overexpression promoter protein expression public health relevance research study response senescence stromelysin 2 transcription factor transcription factor PML tumor ultraviolet

项目摘要

DESCRIPTION (provided by applicant): HDAC7 is a member of the class IIa HDACs that associate with transcription factors including myocyte factor 2 (MEF2) to repress expression of target genes. We show that HDAC7 interacts with PML (promyelocytic leukemia protein). Recent studies in mice have demonstrated a critical role for HDAC7 and PML in the normal development of endothelial cells (ECs). The relationship between HDAC7 and PML is complex. Of primary importance is that the binding of HDAC7 to PML leads to derepression of HDAC7 target genes with clear biological consequences such as altered expression of HDAC7 target genes in the case of endothelial cells. The interaction of HDAC7 and PML also controls the level of PML sumoylation and PML NB formation. All of these effects will be investigated at the mechanistic level with a focus on endothelial cells and their responses to inflammatory stimuli. The specific aims are: 1) To elucidate the mechanisms underlying TNF1-induced accumulation of PML mRNA and protein levels. We will employ chemical inhibitors, siRNA, and dominant-negative expression strategies to delineate the mechanisms controlling TNF1-induced accumulation of PML mRNA, protein levels, PML sumoylation, and formation of PML NBs. 2) To investigate the mechanisms by which HDAC7 mediates TNF1-induced MCP-1 transcription. We hypothesize that TNF1-induced formation of PML NBs sequesters HDAC7, leading to derepression of MMP-10 and MCP-1, both of which are targets of HDAC7. We will test our model using chromatin immunoprecipitation (ChIP), knockdown by siRNA, and transient transfection reporter assays to characterize the MCP-1 promoter. We are also interested in determining whether changes in the histone modifications occur at the MCP-1 promoter upon TNF1 treatment. We will simultane3) To dissect the role of PML and TNF1 in ECs. We will dissect the roles of PML in TNF1-mediated adhesion, migration, proliferation, apoptosis, and remodeling of blood vessels using in vitro and in vivo approaches. In summary, our proposed study is aimed at dissecting the mechanisms and identifying the effectors in cytokine-mediated EC activation. Understanding the pathways that control inflammation, proliferation, apoptosis, and angiogenesis may have therapeutic implications for treating vascular diseases. PUBLIC HEALTH RELEVANCE: Endothelial cells reprogram their transcriptional network in response to cytokine stimulation such as tumor necrosis factor alpha (TNF1), interleukin (IL), lipopolysaccharide (LPS), or interferons (IFNs). Cytokine activation of endothelial cells initiates a series of responses including inflammation, cellular proliferation, apoptosis, and remodeling of blood vessels. The immediate receptors for these cytokines have been intensively studied, whereas the mechanisms underlying transcriptional regulation by downstream effectors remain largely unknown. We have identified histone deacetylase 7 (HDAC7) and promyelocytic leukemia (PML) protein as key mediators in TNF1-induced altered gene expression in human umbilical vascular endothelial cells (HUVECs). We have also shown that PML and HDAC7 play a role in tube formation in HUVECs and human microvascular endothelial cells (HMVECs) in vitro. We will dissect the mechanisms and identify the effectors in TNF1-mediated activation of ECs. Understanding the signaling that control inflammation, proliferation, apoptosis, and angiogenesis may have therapeutic implication in treating vascular diseases.

描述（由申请人提供）：HDAC7是IIA类HDAC的成员，与转录因子相关联，包括肌细胞因子2（MEF2），以抑制靶基因的表达。我们表明HDAC7与PML相互作用（Promyelocytic白血病蛋白）。对小鼠的最新研究表明，HDAC7和PML在内皮细胞（ECS）的正常发育中起着关键作用。 HDAC7和PML之间的关系很复杂。最重要的是，HDAC7与PML的结合导致HDAC7靶基因的压抑具有明显的生物学后果，例如在内皮细胞的情况下，HDAC7靶基因的表达改变了。 HDAC7和PML的相互作用还控制PML Sumoylation和PML NB形成的水平。所有这些影响将在机械水平上研究，重点是内皮细胞及其对炎症刺激的反应。具体目的是：1）阐明TNF1诱导的PML mRNA和蛋白质水平的积累的机制。我们将采用化学抑制剂，siRNA和显性阴性表达策略来描述控制TNF1诱导的PML mRNA积累，蛋白质水平，PML Sumoylation和PML NB的形成的机制。 2）研究HDAC7介导TNF1诱导的MCP-1转录的机制。我们假设TNF1诱导的PML NBS隔离器HDAC7的形成，导致MMP-10和MCP-1的压抑，这两个都是HDAC7的靶标。我们将使用染色质免疫沉淀（CHIP），siRNA敲低以及瞬时转染报告基因测定法测试我们的模型，以表征MCP-1启动子。我们还有兴趣确定TNF1处理后MCP-1启动子发生的组蛋白修饰的变化。我们将同时剖析PML和TNF1在EC中的作用。我们将使用体外和体内方法来剖定PML在TNF1介导的粘附，迁移，增殖，凋亡以及血管重塑的作用。总而言之，我们提出的研究旨在剖析机制并确定细胞因子介导的EC激活中的效应子。了解控制炎症，增殖，凋亡和血管生成的途径可能对治疗血管疾病具有治疗意义。公共卫生相关性：对响应细胞因子刺激的转录网络，例如肿瘤坏死因子α（TNF1），白介素（IL），脂多糖（LPS）或Interferons（IFNS）（IFNS）。内皮细胞的细胞因子激活引发了一系列反应，包括炎症，细胞增殖，凋亡和血管重塑。这些细胞因子的直接受体已经进行了深入的研究，而下游效应子的转录调节的基础机制仍然很大未知。我们已经确定组蛋白脱乙酰基酶7（HDAC7）和前胞细胞白血病（PML）蛋白是TNF1诱导的人脐血管内皮细胞（HUVEC）中基因表达改变的关键介质。我们还表明，PML和HDAC7在体外的HUVEC和人类微血管内皮细胞（HMVEC）中起作用。我们将剖析机制，并确定TNF1介导的EC激活中的效应子。了解控制炎症，增殖，凋亡和血管生成的信号可能对治疗血管疾病具有治疗意义。