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

项目总结 项目3.氧化特定表位在CVD和NASH中的关键作用 脂质过氧化是动脉粥样硬化和NASH的中心事件，导致氧化特异性表位的形成 (OSE)如氧化磷脂(OxPL)、丙二醛(MDA)和称为MAA的复杂的MDA加合物， 我们称之为“氧化特异性表位”(OSE)。它们是促炎性的，并促进慢性 发炎。由于OSE主要是非酶类脂质过氧化的产物，因此其特定的机制 到目前为止还不能中和它们，而且它们在体内在动脉粥样硬化等疾病状态中的实际作用 和纳什都不为人所知。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。 根据我们最近的转基因小鼠的发展，这些转基因小鼠结构性地表达单链抗体， 靶向OxPL--E06-scFv小鼠--或靶向MDA/MAA--IK17-scFv小鼠。在最近出版的和新的 初步数据显示，在食用NASH生产饲料的小鼠中，靶向OxPL降低了 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。 非酒精性脂肪肝患者患心血管疾病的风险增加，并具有共同的危险因素。然而，NAFLD授予 由于已知的共同风险因素，超过这一水平的心血管疾病的额外风险。该项目将检验以下假设： OSE是以前未被认识到的共同风险因素。独一无二的是，这个项目将同时专注于 OSE在动脉粥样硬化和NASH发病机制中的作用并确定共同(或不同)的机制 哪些OSE促进了这些疾病的发生。特异靶1将利用E06-scFv转基因小鼠来研究其作用 OxPL在小鼠动脉粥样硬化和肝脂肪变性/NASH/纤维化模型中的作用及靶向研究 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中和影响动脉粥样硬化形成和肝脏疾病的具体机制。 特异性目标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平行研究的小鼠模型：特异性目标3是OSE在CVD中作用的翻译研究 和NAFLD/NASH，并将寻求确定靶向OxPL和/或MDA/MAA是否不仅可以预防 疾病的进展，但会导致现有疾病的退化。这将使用新的 产生的转基因小鼠有条件地表达针对OSE的抗体，以便增强的滴度可以 在疾病确定之后才能实现。这将使研究能够确定靶向OSE是否具有治疗作用 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和动脉粥样硬化联系起来。因为抗体介导的中和OSE的方法可以靶向 同时，NASH和动脉粥样硬化的发生，这些研究可能导致创新的翻译应用。

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.