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

• 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
• 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

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）：HDAC 7是IIa类HDAC的成员，其与包括肌细胞因子2（MEF 2）在内的转录因子相关以抑制靶基因的表达。我们发现HDAC 7与PML（早幼粒细胞白血病蛋白）相互作用。最近在小鼠中的研究已经证明了HDAC 7和PML在内皮细胞（EC）的正常发育中的关键作用。HDAC 7和PML之间的关系是复杂的。最重要的是HDAC 7与PML的结合导致HDAC 7靶基因的去阻遏，具有明确的生物学后果，例如在内皮细胞的情况下HDAC 7靶基因的表达改变。HDAC 7和PML的相互作用还控制PML类小泛素化和PML NB形成的水平。所有这些作用将在机制水平上进行研究，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TNF 1诱导PML mRNA和蛋白水平积聚的机制。我们将采用化学抑制剂、siRNA和显性负表达策略来描述控制TNF 1诱导的PML mRNA、蛋白水平、PML类小泛素化和PML NB形成的机制。2)研究HDAC 7介导TNF 1诱导的MCP-1转录的机制。我们假设TNF 1诱导的PML NB的形成隔离HDAC 7，导致MMP-10和MCP-1的去抑制，这两者都是HDAC 7的靶点。我们将使用染色质免疫沉淀（ChIP）、siRNA敲低和瞬时转染报告基因测定来测试我们的模型，以表征MCP-1启动子。我们也有兴趣确定组蛋白修饰的变化是否发生在MCP-1启动子TNF-1治疗后。我们将探讨PML和TNF-1在EC中的作用。我们将使用体外和体内的方法来剖析PML在TNF 1介导的粘附、迁移、增殖、凋亡和血管重塑中的作用。总之，我们提出的研究的目的是解剖的机制，并确定在姜黄素介导的EC激活的效应。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：内皮细胞响应于细胞因子刺激（例如肿瘤坏死因子α（TNF 1）、白介素（IL）、脂多糖（LPS）或干扰素（IFN））而重编程其转录网络。内皮细胞的细胞因子活化引发一系列反应，包括炎症、细胞增殖、细胞凋亡和血管重塑。这些细胞因子的直接受体已被深入研究，而下游效应子的转录调控机制仍在很大程度上未知。我们已经确定组蛋白去乙酰化酶7（HDAC 7）和早幼粒细胞白血病（PML）蛋白作为TNF-1诱导的人脐血管内皮细胞（HUVEC）基因表达改变的关键介质。我们还表明，PML和HDAC 7在体外HUVEC和人微血管内皮细胞（HMVEC）中的管形成中发挥作用。我们将剖析的机制，并确定在TNF 1介导的激活内皮细胞的效应。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。