Redox-Control of MMP-1 and Senescence

描述（由申请方提供）：基质金属蛋白酶-1（MMP-1）或间质胶原酶是一种分泌性蛋白，可导致许多年龄相关性退行性疾病的病因。MMP-1在许多癌症中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已被证明在蛋白水解释放和激活生长因子、细胞因子和信号肽中起重要作用，这些生长因子、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的表达中起着重要的作用。本申请旨在确定氧化剂敏感性信号传导网络是否负责MMP-1产生中与年龄相关的增加。我们将测试的假设，即氧化还原激活的Jun-N-末端激酶（JNK）是负责MMP-1表达的年龄依赖性增加，这种氧化还原敏感的信号通路控制活性的Ets-1转录因子和染色质重塑事件，导致增强MMP-1的表达。我们的研究分为四个具体的目标：目标1将确定，1）SA增加MMP-1表达是否对JNK活性的分子或药物抑制敏感，2）JNK激活是否是氧化还原反应性的，3）SA MMP-1表达是否归因于JNK调节磷酸酶的氧化失活;目的2将确定，1）MMP-1启动子的氧化还原响应调节区，其有助于其衰老表达，2）Ets-1募集到MMP-1启动子是否是SA和氧化还原响应的，和3）评估衰老是否导致选择性利用含有MMP-1等位基因的SNP，所述SNP通常与对某些癌症的频率和易感性增加相关;目的3将确定控制衰老相关MMP-1产生的氧化剂敏感性染色质修饰和重塑事件;目标4将使用氧化应激的鼠老化模型确定基于抗氧化剂的疗法是否在以下方面有效：1）降低H2 O2的稳态产生，2）调节激酶信号传导和3）降低MMP的SA表达。我们与衰老生物学、自由基化学和遗传学领域的专家合作，提出了一项解决衰老和肿瘤生物学相关问题的提案，该提案在PA-07-278《衰老细胞的分泌模式》中概述。潜在的总体假设是衰老微环境对退行性疾病的容许性质归因于SA氧化剂产生和异常MMP产生。最后，也许是最重要的，预计我们的研究将从根本上推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过程相关，包括关节炎、动脉粥样硬化、癌症和神经退行性疾病。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，与年龄相关的MMP-1表达增加可以通过抗氧化剂为基础的治疗来预防。对各种抗氧化剂化合物如何阻断基质金属蛋白酶-1产生的基本理解对于设计安全有效的治疗策略以用于未来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.