Targeting NF-kB in Atherosclerosis

描述(申请人提供)：动脉粥样硬化是一种进展缓慢的大中型动脉慢性炎症性疾病，在西方社会导致高死亡率。因此，开发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，将对人类健康保健产生深远的影响。在疾病的每个阶段，发展中的动脉粥样硬化病变中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，从而加剧动脉粥样硬化的形成。与动脉粥样硬化相关的一个主要信号机制是核因子-kB转录因子家族的激活。在动脉粥样硬化斑块中已经证实了活性的核因子-kB，并在体外证实了核因子-kB在促动脉粥样硬化基因表达中的重要性。然而，目前尚不清楚核因子-kB在动脉粥样硬化形成过程中的整体作用是积极的还是消极的，抑制核因子-kB信号在体内对动脉粥样硬化的影响也知之甚少。此外，最近已经描述了导致不同的NF-kB激活的两种主要信号机制，即经典和非典型的NF-kB通路。到目前为止，只有经典途径与动脉粥样硬化的形成有关；然而，新出现的证据有力地支持了非典型途径在动脉粥样硬化发展中的作用。我们的目标是确定抑制NF-：KB信号对动脉粥样硬化发展的整体影响。我们将使用我们开发的新的最先进的药理学和遗传学方法的组合，并通过使用体外和体内动脉粥样硬化模型来实现这一点。这些方法将提供对疾病进展背后的分子和生化机制的更深层次的了解，并将确定在动脉粥样硬化中通过治疗阻断核因子-kB的有效性。我们将检验的假设是：“抑制核因子-kB的激活将防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一问题：(1)确定EC固有的经典和NC核因子-kB信号在动脉粥样硬化形成中的作用；(2)确定经典和NC核因子-kB激活在EC促动脉粥样硬化信号和基因表达中的作用；(3)确定NEMO结合结构域(NBD)在体内对动脉粥样硬化的影响。这些研究将首次直接探讨非典型的核因子-kB途径在动脉粥样硬化形成中的作用。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地阻断经典的NF-kB信号可以防止动脉粥样硬化的发展。成功实现我们的目标将为靶向经典和非典型的核因子-kB信号在动脉粥样硬化患者中的潜在治疗价值提供至关重要的新见解。因此，我们的建议意义重大，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.