Targeting NF-kB in Atherosclerosis

描述（由申请人提供）：动脉粥样硬化是一种缓慢进展的中大动脉慢性炎症性疾病，在西方社会是造成高死亡率的原因。因此，制定有效的策略来预防或治疗动脉粥样硬化是一个非常重要的目标，将深刻影响人类的健康保健。在疾病的每个阶段，发展中的动脉粥样硬化病变内的促动脉粥样硬化因子激活关键细胞类型，包括内皮细胞（EC）、平滑肌细胞和巨噬细胞。当被激活时，这些细胞表达细胞因子、趋化因子、细胞表面分子和酶，加剧动脉粥样硬化的发生。与动脉粥样硬化相关的主要信号机制是NF-kB家族转录因子的激活。活性NF-kB已在动脉粥样硬化斑块中得到证实，NF-kB对促动脉粥样硬化基因表达的重要性已在体外得到证实。然而，目前尚不清楚NF-:kB在动脉粥样硬化过程中是否起着积极或消极的作用，并且体内抑制NF-kB信号传导对动脉粥样硬化的影响尚不清楚。此外，最近已经描述了导致不同NF-kB物种激活的两种主要信号机制，称为经典和非典型NF-kB途径。迄今为止，只有经典途径与动脉粥样硬化有关；然而，新出现的证据强烈支持非规范途径在动脉粥样硬化发展过程中的作用。我们的目标是确定抑制NF-:kB信号对动脉粥样硬化发展的总体影响。我们将结合我们开发的最先进的药理学和遗传学方法，并采用体外和体内动脉粥样硬化模型来实现这一目标。这些方法将提供对疾病进展的分子和生化机制的更深入理解，并将确定在动脉粥样硬化中治疗阻断NF-kB的有效性。我们将验证的假设是：“抑制NF-kB的激活将阻止动脉粥样硬化的发展”。我们将通过以下三个具体目标来解决这个问题：(1)定义ec内在经典和NC NF-kB信号在动脉粥样硬化过程中的作用；(2)确定经典和NC NF-kB激活在EC中促动脉粥样硬化信号传导和基因表达中的作用；(3)在体内确定NEMO结合域（NBD）肽对动脉粥样硬化的影响。这些研究将首次直接探讨非规范NF-kB通路在动脉粥样硬化中的作用。此外，NBD肽从未在动脉粥样硬化模型中进行过体内测试，我们预测这种独特的方法将证明系统性阻断经典NF-kB信号传导可阻止动脉粥样硬化的发展。成功实现我们的目标将为动脉粥样硬化患者靶向经典和非典型NF-kB信号的潜在治疗价值提供重要的新见解。因此，我们的建议非常重要，并广泛影响人类健康关注的一个关键领域。

DESCRIPTION (provided by applicant): Atherosclerosis is a slowly progressing chronic inflammatory disease of the medium and large arteries that is responsible for high mortality rates in western society. Consequently, developing effective strategies to prevent or cure atherosclerosis is a highly significant objective that will profoundly impact human health care. At each stage of the disease, pro-atherogenic factors within the developing atherosclerotic lesion activate crucial cell types including endothelial cells (EC), smooth muscle cells and macrophages. When activated, these cells express cytokines, chemokines, cell surface molecules and enzymes that exacerbate atherogenesis. A major signaling mechanism associated with atherosclerosis is activation of the NF-kB family of transcription factors. Active NF-kB has been demonstrated in atherosclerotic plaques, and the importance of NF-kB for pro-atherogenic gene expression has been established in vitro. However, it remains unclear whether NF-:kB plays an overall positive or negative role during atherogenesis, and the effects of inhibiting NF-kB signaling on atherosclerosis in vivo are poorly understood. Furthermore, two major signaling mechanisms leading to the activation of distinct NF-kB species, named the classical and non-canonical NF-kB pathways, have been described recently. To date, only the classical pathway has been implicated in atherogenesis; however, emerging evidence strongly supports a role for the non-canonical pathway during the development of atherosclerosis. Our goal is to determine the overall effects of inhibiting NF-:kB signaling on the development of atherosclerosis. We will accomplish this using a combination of novel state-of-the-art pharmacological and genetic approaches that we have developed and by employing both in vitro and in vivo models of atherosclerosis. These approaches will provide both a deeper understanding of the molecular and biochemical mechanisms underlying disease progression and will determine the effectiveness of therapeutically blocking NF-kB in atherosclerosis. The hypothesis we will test is: "Inhibiting NF-kB activation will prevent the development of atherosclerosis". We will address this by pursuing the following three specific aims: (1) To define the roles of EC-intrinsic classical and NC NF-kB signaling during atherogenesis; (2) To determine the role of classical and NC NF-kB activation in pro-atherogenic signaling and gene expression in EC; (3) To determine the effects of the NEMO binding domain (NBD) peptide on atherosclerosis in vivo. These studies will directly address the role of the non-canonical NF-kB pathway in atherogenesis for the first time. Furthermore, the NBD peptide has never been tested in vivo in models of atherosclerosis and we predict that this unique approach will demonstrate that systemically blocking classical NF-kB signaling prevents the development of atherosclerosis. Successfully accomplishing our aims will provide crucial novel insight into the potential therapeutic value of targeting classical and non-canonical NF-kB signaling in atherosclerosis patients. Consequently, our proposal is highly significant and broadly impacts a critical area of human health concern.