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关节炎和其他几种慢性炎症疾病中，炎症性细胞因子IL-1水平高，与疾病的严重程度相关，而抗炎IL-4水平则低或不可检测。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和肺泡骨破坏的丧失。 MMP-3具有广泛的底物特异性，可以激活其他Pro-MMP。在患病部位的水平增加中，与严重程度和进展相关。 IL-4抑制IL-1诱导的MMP-3在牙周炎患者中分离的人牙龈成纤维细胞中的表达。将父亲位点（Stromelysin IL-1响应元件）确定为抑制MMP-3诱导IL-1诱导的阻遏元件。它也是影响转录的常见5​​T/6T多态性的位点，5T等位基因与较高的表达相关。该部位的基因型也与MMP-3的组织水平以及包括牙周炎在内的多种疾病的易感性或严重程度有关。在心血管疾病（与牙周炎有关的疾病）中，高表达5T等位基因的纯合性与心肌梗塞和动脉瘤有关，而低表达6T等位基因与动脉粥样硬化有关。因此，很明显，必须严格控制该基因的调节以维持正确的组织稳态，并且了解这些控制机制对于多种病理学很重要。 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.染色质免疫沉淀（CHIP）测定将用于确定在几种细胞类型和条件下在5T和6T位点的体内结合的转录因子，并且还将比较与副因素的相互作用。还将测试通过小疗法RNA（siRNA）和/或使用化学抑制剂对每个因素的敲除效果。 2）确定IL-4抑制MMP-3表达的机理，特别集中在STAT6，P300/CBP和/或蛋白质磷酸酶2A（PP2A）的作用上。这些研究将利用瞬时转染，siRNA和实时PCR。在此过程中，我们希望获得有关MMP-3调节涉及的复杂基因调节机制的信息，但也继续为PCOM的研究环境以及对整个研究的学生培训，尤其是分子生物学的培训。牙周炎是美国成人牙齿脱落的最常见原因，并有助于其他几种疾病的发展。 MMP-3表达与牙周炎的支撑结构的破坏有关，并且在其他慢性炎症的疾病中也起作用。 MMP-3启动子中的一个常见启动子多态性已被证明会影响转录并具有许多疾病关联（包括牙周炎），这表明MMP-3过多或太少MMP-3可能会产生病理后果，并且必须严格控制该基因以维持适当的组织稳态。我们对这些转录控制机制以及它们有时失败的方式增加了，对于提高我们对各种病理的理解至关重要。

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.

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

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