Redox-Control of MMP-1 and Senescence

描述（由申请人提供）：基质金属蛋白酶-1 （MMP-1）或间质胶原酶是一种分泌蛋白，与许多年龄相关的退行性疾病的病因有关。在许多癌症中，MMP-1的异常表达也与疾病的严重程度和患者预后不良有关。近年来，MMP-1已被证明在生长因子、细胞因子和信号肽的蛋白水解释放和激活中发挥重要作用，这些细胞因子和信号肽也具有调节衰老微环境的潜力。控制衰老相关（SA） MMP-1表达的调控机制尚不清楚。我们的研究结果表明，氧化剂在调节MMP-1表达中起重要作用。本应用旨在确定氧化敏感信号网络是否与年龄相关的MMP-1生成增加有关。我们将验证以下假设：jun - n-末端激酶（JNK）的氧化还原激活导致MMP-1表达的年龄依赖性增加，这种氧化还原敏感信号通路控制Ets-1转录因子的活性和导致MMP-1表达增强的染色质重塑事件。我们的研究分为四个具体目的：目的1将确定，1)SA MMP-1表达的增加是否对JNK活性的分子或药物抑制敏感，2)JNK激活是否具有氧化还原反应，3)SA MMP-1表达是否归因于JNK调节磷酸酶的氧化失活；Aim 2将确定，1)MMP-1启动子的氧化还原反应调控区域，该区域有助于其衰老表达，2)Ets-1募集到MMP-1启动子是否具有SA和氧化还原反应，以及3)评估衰老是否导致选择性利用含有MMP-1等位基因的SNP，该等位基因通常与某些癌症的频率和易感性增加有关；目的3将确定氧化敏感性染色质修饰和重塑事件，控制衰老相关的MMP-1的产生；Aim 4将使用小鼠氧化应激衰老模型来确定基于抗氧化的治疗是否有效，1)减少H2O2的稳态产生，2)调节激酶信号传导，3)降低MMPs的SA表达。我们已经与衰老生物学、自由基化学和遗传学领域的专家结盟，制定了一项提案，解决与衰老和肿瘤生物学相关的相关问题，并在PA-07-278中概述，衰老细胞的分泌模式。潜在的总体假设是，衰老微环境对退行性疾病的宽容性质归因于SA氧化剂的产生和MMP的异常产生。最后，也许最重要的是，我们的研究将从根本上促进我们对自由基对许多SA疾病的贡献的理解。公共卫生相关性。衰老过程与许多退行性疾病有关，包括关节炎、动脉粥样硬化、癌症和神经变性。与许多这些疾病过程相关的是基质金属蛋白酶-1的不受控制的表达。我们的实验室已经确定，年龄相关的MMP-1表达增加可以通过基于抗氧化剂的治疗来预防。了解各种抗氧化化合物如何阻断基质金属蛋白酶-1的产生，对于设计安全有效的治疗策略以治疗许多与年龄相关的退行性疾病至关重要。

DESCRIPTION (provided by applicant): Matrix metalloproteinase-1 (MMP-1) or interstitial collagenase is a secreted protein which contributes to the etiology of many age-related degenerative diseases. Aberrant expression of MMP-1 in many cancers is also linked to both severity of disease and poor patient outcome. In recent years MMP-1 has been shown to play a prominent role in the proteolytic release and activation of growth factors, cytokines and signaling peptides which also have the potential to modulate the senescent microenvironment. The regulatory mechanisms that control senescence associated (SA) MMP-1 expression are unknown. Our findings indicate that oxidants play an important role in regulating MMP-1 expression. This application seeks to define whether oxidant-sensitive signaling networks are responsible for age associated increases in MMP-1 production. We will test the hypotheses that redox activation of Jun-N-Terminal Kinase (JNK) is responsible for the age-dependent increases in MMP-1 expression and that this redox-sensitive signaling pathway controls the activity of the Ets-1 transcription factor and chromatin remodeling events that lead to enhanced MMP-1 expression. Our studies are divided into four specific aims: Aim 1 will determine,1) if the SA increase in MMP-1 expression is sensitive to molecular or pharmacologic inhibition of JNK activity, 2) whether JNK activation is redox-responsive and 3) if SA MMP-1 expression is attributed to oxidative-inactivation of JNK regulatory phosphatases; Aim 2 will determine, 1) the redox responsive regulatory regions of the MMP-1 promoter that contribute to its senescence expression, 2) whether Ets-1 recruitment to the MMP-1 promoter is SA and redox- responsive, and 3) evaluate if senescence leads to the selective utilization of the SNP containing MMP-1 allele that is commonly associated with enhanced frequency and susceptibility to certain cancers; Aim 3 will determine the oxidant-sensitive chromatin modifying and remodeling events that control senescence associated MMP-1 production; and Aim 4 will determine, using a murine aging model of oxidative stress, if antioxidant based therapies are effective in, 1) decreasing the steady state production of H2O2, 2) modulating kinase signaling and 3) decreasing the SA expression of MMPs. We have aligned ourselves with experts in the fields of aging biology, free radical chemistry and genetics to develop a proposal which tackles relevant issues related to both aging and tumor biology and that were outlined in the PA-07-278, The Secretory Pattern of Senescent Cells. The underlying overall hypothesis is that the permissive nature of the senescent microenvironment to degenerative disease is attributed to SA oxidant production and aberrant MMP production. Finally, and perhaps most importantly, it is anticipated that our studies will fundamentally advance our understanding of the contribution of free radicals to many SA diseases. Public Health Relevance. The process of aging is associated with many degenerative disease processes including arthritis, atherosclerosis, cancer and neurodegeneration. Associated with many of these disease processes is the uncontrolled expression of matrix metalloproteinase-1. Our laboratory has established that the age-associated increases in MMP-1 expression can be prevented by antioxidant-based therapies. A fundamental understanding of how the various antioxidant compounds block matrix metalloproteinase-1 production is essential for the design of safe and effective therapeutic strategies for the future treatment of many age associated degenerative diseases.