Transcriptional Regulation of Matrix Metalloproteinase-3 (MMP-3)

基质金属蛋白酶 3 (MMP-3) 的转录调控

• 批准号: 7303944
• 负责人: RUTH C BORGHAEI
• 金额: $ 22.5万
• 依托单位: PHILADELPHIA COLLEGE OF OSTEOPATHIC MED
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-28 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7303944
• 关键词: Adult; Adverse effects; Affect; Alleles; Aneurysm; Animal Model; Anti-Inflammatory Agents; Anti-inflammatory; Atherosclerosis; Bacteria; Be++ element; Beryllium; Binding; Biological Assay; COS Cells; Cardiovascular Diseases; Cells; Chemicals; Chromatin; Chronic; Complex; Condition; Development; Diabetes Mellitus; Disease; Disease Association; EP300 gene; Elements; Environment; Equilibrium; Fibroblasts; Fright; Gene Expression; Gene Expression Regulation; Genes; Genetic Enhancer Element; Genetic Polymorphism; Genetic Transcription; Genotype; Gingiva; Homeostasis; Human; IL4 gene; Indium; Inflammation; Inflammatory; Integration Host Factors; Interleukin-1; Interleukin-4; Interleukins; Joints; Link; Low Birth Weight Infant; Matrix Metalloproteinases; Messenger RNA; Modeling; Molecular; Molecular Biology; Myocardial Infarction; NF-kappa B; Numbers; Other Therapy; Pathology; Patients; Periodontitis; Peroxisome Proliferator-Activated Receptors; Plasmids; Play; Polymerase Chain Reaction; Predisposition; Production; Progress Reports; Protein phosphatase; Proteins; Publications; Recombinants; Regulation; Regulator Genes; Relative (related person); Research; Rheumatoid Arthritis; Risk; Role; STAT6 gene; Series; Severities; Severity of illness; Site; Small Interfering RNA; Stenosis; Stromelysin 1; Structure; Students; Substrate Specificity; TNFRSF5 gene; Testing; Therapeutic; Time; Tissue Sample; Tissues; Tooth Loss; Training; Transcription Factor AP-1; Transcriptional Activation; Transcriptional Regulation; Transfection; Tumor Necrosis Factor-alpha; Tumor Necrosis Factors; Up-Regulation; alveolar bone; base; cell type; chromatin immunoprecipitation; cytokine; dimer; gene therapy; human TNF protein; improved; in vivo; inhibitor/antagonist; knock-down; mutant; p65; promoter; research study; transcription factor; zinc-binding protein

DESCRIPTION (provided by applicant): In periodontitis, as well as rheumatoid arthritis and several other chronic inflammatory conditions, levels of inflammatory cytokine IL-1 are high and correlate with disease severity, while levels of anti-inflammatory IL-4 are low or undetectable. Furthermore, decreasing levels of IL-4 are correlated with increasing severity. IL-1 contributes to loss of attachment and alveolar bone destruction by increasing expression of matrix metalloproteinases. MMP-3 has broad substrate specificity and can activate other pro-MMP. It is found in increased levels in diseased sites, correlated with severity and progression. IL-4 suppresses the IL-1 induced expression of MMP-3 in human gingival fibroblasts isolated from patients with periodontitis. The SIRE site (stromelysin IL-1 responsive element) was identified as a repressor element involved in suppressing the IL-1 induction of MMP-3. It is also the site of a common 5T/6T polymorphism that affects transcription, with the 5T allele associated with higher levels of expression. Genotype at this site has also been linked to tissue levels of MMP-3 and to susceptibility or severity of a number of diseases, including periodontitis. In cardiovascular disease (a condition associated with periodontitis), homozygosity for the higher- expressing 5T allele is associated with myocardial infarction and aneurysm, while the lower- expressing 6T allele is associated with atherosclerosis. It is therefore clear that regulation of this gene must be tightly controlled to maintain correct tissue homeostasis, and that understanding these control mechanisms is important for a variety of pathologies. Transcription factors ZBP-89 and NFkB bind to the SIRE site, and evidence suggests that ZBP-89 activates transcription, especially from the 5T site, while NF-?B represses from both 5T and 6T The specific aims of this proposal are to: 1) Study the roles of transcription factors interacting with the MMP-3 promoter, focusing in particular on the polymorphic SIRE site. The chromatin immunoprecipitation (ChIP) assay will be used to determine which transcription factor(s) bind in vivo at the 5T and 6T sites in several cell types and conditions, and interactions with co-factors will also be compared. The effects of knock-down of each of the factors by small-interfering RNA (siRNA) and/or by using chemical inhibitors will also be tested. 2) Determine the mechanism of suppression of MMP-3 expression by IL-4, concentrating in particular on the roles of STAT6, p300/CBP and/or protein phosphatase 2A (PP2A). These studies will utilize transient transfection, siRNA and real-time PCR. In doing so, we hope to gain information about complex gene regulatory mechanisms involved in MMP-3 regulation, but also continue to contribute to the research environment at PCOM and to the training of its students in research in general, and molecular biology in particular. Periodontitis is the most common cause of adult tooth loss in the U.S., and contributes to the development of several other diseases. MMP-3 expression is associated with destruction of support structures in periodontitis, and also plays a role in other diseases with chronic inflammation. A common promoter polymorphism in the MMP-3 promoter has been shown to affect transcription and to have a number of disease associations (including with periodontitis), suggesting that either too much or too little MMP-3 can have pathological consequences and that expression of this gene must be tightly controlled in order to maintain proper tissue homeostasis. Our increased understanding of these transcriptional control mechanisms, and how they sometimes fail, will be important to improve our understanding of a variety of pathologies.

描述（由申请人提供）：在牙周炎以及类风湿性关节炎和几种其它慢性炎性病症中，炎性细胞因子IL-1的水平高并且与疾病严重性相关，而抗炎性IL-4的水平低或检测不到。此外，IL-4水平的降低与严重程度的增加相关。IL-1通过增加基质金属蛋白酶的表达而导致附着丧失和牙槽骨破坏。MMP-3具有广泛的底物特异性，并可激活其他MMP前体。它在患病部位的水平增加，与严重程度和进展相关。IL-4抑制IL-1诱导的牙周炎患者牙龈成纤维细胞MMP-3的表达SIRE位点（基质溶解素IL-1反应元件）被鉴定为参与抑制MMP-3的IL-1诱导的阻遏元件。它也是影响转录的常见5 T/6 T多态性的位点，5 T等位基因与更高的表达水平相关。该位点的基因型也与MMP-3的组织水平以及许多疾病（包括牙周炎）的易感性或严重性有关。在心血管疾病（与牙周炎相关的病症）中，高表达的5 T等位基因的纯合性与心肌梗死和动脉瘤相关，而低表达的6 T等位基因与动脉粥样硬化相关.因此，很明显，该基因的调控必须严格控制，以保持正确的组织稳态，了解这些控制机制是重要的各种病理。转录因子ZBP-89和NF-κ B结合SIRE位点，有证据表明ZBP-89激活转录，特别是从5 T位点，而NF-？B从5 T和6 T两方面抑制该提议的具体目的是：1）研究与MMP-3启动子相互作用的转录因子的作用，特别关注多态性SIRE位点。染色质免疫沉淀（ChIP）试验将用于确定在几种细胞类型和条件下，哪些转录因子在体内结合在5 T和6 T位点，还将比较与辅因子的相互作用。还将测试通过小干扰RNA（siRNA）和/或通过使用化学抑制剂敲低每种因子的效果。2)确定IL-4抑制MMP-3表达的机制，特别关注STAT 6、p300/CBP和/或蛋白磷酸酶2A（PP 2A）的作用。这些研究将利用瞬时转染、siRNA和实时PCR。通过这样做，我们希望获得有关MMP-3调控中涉及的复杂基因调控机制的信息，但也继续为PCOM的研究环境和学生在一般研究中的培训做出贡献，特别是分子生物学。牙周炎是美国成年人牙齿脱落的最常见原因，并导致其他几种疾病的发展。MMP-3的表达与牙周炎中支持结构的破坏有关，并且在其他慢性炎症疾病中也起作用。MMP-3启动子中常见的启动子多态性已被证明影响转录并具有许多疾病相关性（包括牙周炎），这表明MMP-3过多或过少都可能具有病理后果，并且必须严格控制该基因的表达以维持适当的组织稳态。我们对这些转录控制机制的理解，以及它们有时是如何失败的，对于提高我们对各种病理学的理解将是重要的。

项目成果

IL-4 inhibition of IL-1 induced Matrix metalloproteinase-3 (MMP-3) expression in human fibroblasts involves decreased AP-1 activation via negative crosstalk involving of Jun N-terminal kinase (JNK).

• DOI: 10.1016/j.yexcr.2013.04.010
• 发表时间: 2013-06-10
• 期刊: EXPERIMENTAL CELL RESEARCH
• 影响因子: 3.7
• 作者: Chambers, Mariah;Kirkpatrick, Garrett;Evans, Michel;Gorski, Grzegorz;Foster, Sara;Borghaei, CRuth C.
• 通讯作者: Borghaei, CRuth C.

Expression of transcription factor zinc-binding protein-89 (ZBP-89) is inhibited by inflammatory cytokines.

• DOI: 10.2147/plmi.s6249
• 发表时间: 2009-08-01
• 期刊: Pathology and laboratory medicine international
• 影响因子: 0.2
• 作者: Borghaei RC;Chambers M
• 通讯作者: Chambers M

Interleukin-4 inhibition of interleukin-1-induced expression of matrix metalloproteinase-3 (MMP-3) is independent of lipoxygenase and PPARgamma activation in human gingival fibroblasts.

白细胞介素 4 对白细胞介素 1 诱导的基质金属蛋白酶 3 (MMP-3) 表达的抑制与人牙龈成纤维细胞中脂氧合酶和 PPARgamma 的激活无关。

• DOI: 10.1186/1471-2199-8-12
• 发表时间: 2007
• 期刊: BMC molecular biology
• 作者: Stewart,Denise;Javadi,Masoud;Chambers,Mariah;Gunsolly,Chad;Gorski,Grzegorz;Borghaei,RuthC
• 通讯作者: Borghaei,RuthC