Pivotal Role of Oxidation-specific Epitopes in CVD and NASH.

项目概要 项目 3. 氧化特异性表位在 CVD 和 NASH 中的关键作用 脂质过氧化是动脉粥样硬化和 NASH 的核心事件，导致氧化特异性表位的形成 (OSE)，例如氧化磷脂 (OxPL)、丙二醛 (MDA) 和称为 MAA 的复杂 MDA 加合物， 我们将其称为“氧化特异性表位”（OSE）。它们具有促炎性并促进慢性 炎症。由于 OSE 大部分是非酶脂质过氧化的产物，因此具体机制 迄今为止，还无法中和它们，而它们在动脉粥样硬化等疾病状态中的体内实际作用 和 NASH 未知。项目 3 的重点是了解 OSE 在动脉粥样硬化和 NASH 以及它们导致这些疾病的常见（或独特）机制。现在这是可行的 基于我们最近开发的转基因小鼠，其组成型表达单链抗体 靶向 OxPL（E06-scFv 小鼠）或靶向 MDA/MAA（IK17-scFv 小鼠）。在最近发布的和新的 初步数据表明，在食用产生 NASH 的饮食的小鼠中，针对 OxPL 可以降低 动脉粥样硬化和 NASH。初步研究表明，靶向 MDA/MAA 也可以减少 NASH 模型中动脉粥样硬化和肝脏炎症的进展。 NAFLD 患者患 CVD 的风险增加，并且具有共同的危险因素。然而，NAFLD 赋予 由于已知的共同风险因素，CVD 的额外风险高于此。该项目将测试以下假设： OSE 是一种以前未被认识到的常见风险因素。独特的是，该项目将同时关注 OSE 在动脉粥样硬化和 NASH 发病机制中的作用，并通过以下方式定义常见（或不同）机制 其中 OSE 会促进这些疾病。具体目标1将使用E06-scFv转基因小鼠来研究 OxPL 在小鼠模型中动脉粥样硬化和肝脂肪变性/NASH/纤维化中的作用，并确定是否针对 OxPL 可以同时减轻动脉和肝脏的疾病负担。我们将使用 E06-scFv 小鼠 研究中和 OxPL 影响动脉粥样硬化和肝脏疾病的具体机制。 具体目标 2 将研究 MDA/MAA 在动脉粥样硬化和肝脂肪变性/NASH/纤维化中的作用 与目标 1 并行研究的小鼠模型。具体目标 3 是 OSE 在 CVD 中作用的转化研究 和 NAFLD/NASH，并将寻求确定针对 OxPL 和/或 MDA/MAA 是否不仅可以预防 疾病进展但导致现有疾病消退。这将使用新来完成 产生的转基因小鼠有条件地表达针对 OSE 的抗体，从而提高滴度 疾病确立后才能实现。这将使研究能够确定靶向 OSE 是否具有治疗作用 并减轻疾病负担。这些研究应该提供对新的常见风险因素的理解 将 NASH 与动脉粥样硬化联系起来。因为抗体介导的中和 OSE 方法可以靶向 NASH 和动脉粥样硬化同时发生，这些研究可能会带来创新的转化应用。

PROJECT SUMMARY Project 3. Pivotal Role of Oxidation-specific Epitopes in CVD and NASH Lipid peroxidation, a central event in atherogenesis and NASH, results in formation of oxidation-specific epitopes (OSE) such as oxidized phospholipids (OxPL), malondialdehyde (MDA) and complex MDA adducts termed MAA, which we termed “oxidation-specific-epitopes” (OSE). They are proinflammatory and promote chronic inflammation. Because OSE are mostly products of non-enzymatic lipid peroxidation, mechanisms to specifically neutralize them were unavailable until now and their actual roles in vivo in disease states such as atherosclerosis and NASH are unknown. Project 3 is focused on understanding the role of OSE in both atherosclerosis and NASH and the common (or unique) mechanisms by which they contribute to these diseases. This is now feasible based on our recent development of transgenic mice that constitutively express single chain antibodies that target OxPL--the E06-scFv mice—or target MDA/MAA—the IK17-scFv mice. In recently published and new preliminary data we demonstrate that targeting OxPL in mice consuming NASH producing diets decreases both atherosclerosis and NASH. Preliminary studies demonstrate that targeting of MDA/MAA can also reduce the progression of atherosclerosis and hepatic inflammation in a NASH model. Patients with NAFLD are at increased risk for CVD, and share common risk factors. However, NAFLD confers additional risk for CVD above that due to known shared risk factors. This Project will test the hypothesis that OSE are a previously unrecognized common risk factor. Uniquely, this project will simultaneously focus on the role of OSE in the pathogenesis of atherosclerosis and NASH and define common (or distinct) mechanisms by which OSE promote these diseases. Specific Aim 1 will use the E06-scFv transgenic mice to study the Role of OxPL in Atherosclerosis and Hepatic Steatosis/NASH/Fibrosis in mouse models and determine if targeting OxPL can simultaneously reduce disease burden in both the artery and liver. We will use the E06-scFv mice to investigate the specific mechanisms by which neutralization of OxPL impacts atherogenesis and liver disease. Specific Aim 2 will study the Role of MDA/MAA in Atherosclerosis and Hepatic Steatosis/NASH/ Fibrosis in mouse models in parallel studies to Aim 1. Specific Aim 3 are Translational Studies of the Role of OSE in CVD and NAFLD/NASH and will seek to determine if targeting OxPL and/or MDA/MAA can not only prevent progression of disease but cause regression of existing disease. This will be accomplished using newly generated transgenic mice that conditionally express the antibodies targeting OSE so that enhanced titers can be achieved after disease is established. This will allow studies to determine if targeting OSE can be therapeutic and reduce disease burden. These studies should provide an understanding of novel common risk factors connecting NASH and atherogenesis. Because an antibody-mediated approach to neutralize OSE could target both NASH and atherogenesis simultaneously, these studies may lead to innovative translational applications.