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

项目摘要 项目3。氧化特异性表位在CVD和NASH中的作用 脂质过氧化是动脉粥样硬化和NASH的中心事件，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， 我们称之为“氧化特异性表位”（OSE）。它们是促炎性的， 炎症由于OSE主要是非酶脂质过氧化的产物， 直到现在还不能中和它们，它们在诸如动脉粥样硬化的疾病状态中的体内实际作用 和NASH是未知的。项目3的重点是了解OSE在动脉粥样硬化和 NASH及其导致这些疾病的共同（或独特）机制。这现在是可行的 基于我们最近开发的组成型表达单链抗体的转基因小鼠， 靶向OxPL-E06-scFv小鼠-或靶向MDA/MAA-IK 17-scFv小鼠。在最近出版的和新的 我们的初步数据表明，在消耗NASH产生饮食的小鼠中靶向OxPL， 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患者发生CVD的风险增加，并有共同的风险因素。然而，NAFLD赋予 CVD的额外风险高于已知的共同风险因素。本项目将检验以下假设： OSE是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共同（或不同）机制： 哪些OSE促进了这些疾病。具体目标1将使用E06-scFv转基因小鼠来研究E06-scFv的作用。 OxPL在小鼠模型中的动脉粥样硬化和肝脂肪变性/NASH/纤维化中的作用，并确定是否靶向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OxPL的中和作用影响动脉粥样硬化形成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， 小鼠模型与目标1平行研究。具体目标3是OSE在CVD中作用的转化研究 和NAFLD/NASH，并将寻求确定靶向OxPL和/或MDA/MAA是否不仅可以预防 疾病进展，但导致现有疾病的消退。这将使用新的 产生条件性表达靶向OSE的抗体的转基因小鼠， 在疾病发生后，这将允许研究确定靶向OSE是否可以治疗 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 NASH和动脉粥样硬化之间的联系因为抗体介导的中和OSE的方法可以靶向 NASH和动脉粥样硬化的同时，这些研究可能导致创新的转化应用。

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.