Unraveling ApoE4 Promotion of Cardiometabolic Disease

揭示 ApoE4 对心血管代谢疾病的促进作用

基本信息

批准号：
10192811
负责人：
PHILIP W SHAUL
金额：
$ 56.9万
依托单位：
UT SOUTHWESTERN MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-07-01 至 2024-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10192811
关键词：
Address Adhesions Amino Acids Aorta Apolipoprotein E Atherosclerosis Beta Cell Blood Platelets Blood Vessels Blood coagulation Cardiometabolic Disease Cardiovascular Diseases Catalytic Domain Cell Culture Techniques Cell Line Co-Immunoprecipitations Coronary Arteriosclerosis DAB2 gene Data Data Set Development Endothelial Cells Endothelium Enzyme Activation Genes Genetic Goals Health Hepatocyte Human Individual Insulin Insulin Resistance Intervention LDL-Receptor Related Protein 1 Ligands Link Lipids Liver MAPK8 gene Mass Spectrum Analysis Mediating Metabolic Metabolic Diseases Mus Muscle Nitric Oxide Synthase Non-Insulin-Dependent Diabetes Mellitus Observational Study Pathogenesis Population Process Protein Isoforms Protein Phosphatase 2A Regulatory Subunit PR53 Protein Subunits Protein phosphatase Proteins Research Risk SP600125 Signal Transduction Skeletal Muscle Testing Thrombosis Validation Vascular Diseases Venous Thrombosis Work apolipoprotein E receptor 2 apolipoprotein E-3 apolipoprotein E-4 base blood glucose regulation cardiometabolism cardiovascular disorder risk cohort genetic variant genome wide association study in vivo inhibitor/antagonist lipid transport monocyte novel therapeutics pre-clinical preservation prevent programs recruit scaffold von Willebrand Factor

项目摘要

Project Summary/Abstract Compared with individuals expressing the most common genetic variant of apolipoprotein E (apoE), apoE3, those harboring apoE4 are at increased risk of both cardiovascular disease and type 2 diabetes mellitus (T2DM). Although apoE classically participates in lipid transport, the basis for apoE4-associated risk remains unclear, and there are no interventions available to break the link between apoE4 and vascular and metabolic disorders. We recently discovered that the insulin resistance observed in mice expressing human apoE4 is driven by skeletal muscle insulin resistance mediated by the apoE receptor apoER2 in endothelial cells. Studies in culture showed that the underlying mechanism is apoE4-apoER2-induced inhibition of endothelial cell insulin transport, which has a major impact on insulin action in muscle. In other recent work we determined that thrombosis is markedly more severe in apoE4- versus apoE3-expressing mice. Cell culture studies further indicated that this may be due to apoE4 promotion of von Willebrand factor (vWF) secretion by endothelial cells. The Overall Goal of the proposed research is to elucidate how apoE4 actions on endothelium influence glucose homeostasis and vascular disease pathogenesis. Aim 1 will determine how apoE4 causes insulin resistance. A non-biased query of apoER2-interacting proteins and ensuing studies of insulin transport in cultured endothelial cells suggest involvement of Dab2-interacting protein (DAB2IP) and JIP1, which both modulate JNK signaling. Using endothelial cell-specific deletion in apoE3- and apoE4- expressing mice, we will test the hypothesis that apoE4 promotes insulin resistance via endothelial apoER2 through DAB2IP- and JIP1-dependent mechanisms. Aim 2 will determine how apoE4 promotes atherosclerosis and thrombosis. Studies of proteins interacting with apoER2 in endothelium and apoE4 enhancement of endothelial cell vWF secretion and monocyte adhesion indicate possible participation of the protein phosphatase PP2A. In apoE3- and apoE4-expressing mice we will test the hypothesis that apoE4 promotes atherosclerosis and thrombosis via endothelial apoER2-related activation of PP2A. In Aims 1 and 2, we will additionally use GWAS-based strategies to determine if the operative genes in mice are modifier genes influencing apoE4-related risk of metabolic and vascular disease in humans. In Aim 3 we will devise interventions against apoE4 and its mechanisms of action to preserve metabolic and vascular health. In apoE4 mice the effect of JNK inhibition on insulin resistance and the effect of PP2A inhibition on atherosclerosis and thrombosis will be evaluated. We will also use base editing to genetically correct apoE4 to apoE3 in vivo in the hepatocytes of apoE4 mice. Editing efficiency and impact on apoE4-related insulin resistance, atherosclerosis and thrombosis will be evaluated. By accomplishing these Aims and unraveling how apoE4 promotes cardiometabolic disease, it is expected that novel therapies can then be developed to neutralize an important genetic contribution to metabolic and vascular disease in 15 to 20% of the population.

项目概要/摘要与表达最常见的载脂蛋白 E (apoE) 遗传变异的个体相比， apoE3 和含有 apoE4 的人患心血管疾病和 2 型糖尿病的风险增加糖尿病（T2DM）。尽管 apoE 通常参与脂质转运，但 apoE4 相关风险的基础仍不清楚，并且没有可用的干预措施来打破 apoE4 与血管和血管之间的联系代谢紊乱。我们最近发现在表达人类基因的小鼠中观察到胰岛素抵抗 apoE4 是由内皮细胞中的 apoE 受体 apoER2 介导的骨骼肌胰岛素抵抗驱动的细胞。培养研究表明，潜在机制是 apoE4-apoER2 诱导的抑制内皮细胞胰岛素转运，对肌肉中的胰岛素作用有重大影响。在最近的其他工作中我们确定表达 apoE4 的小鼠与表达 apoE3 的小鼠相比，血栓形成明显更严重。细胞培养研究进一步表明，这可能是由于 apoE4 促进血管性血友病因子 (vWF) 由内皮细胞分泌。拟议研究的总体目标是阐明 apoE4 如何发挥作用对内皮细胞影响葡萄糖稳态和血管疾病发病机制。目标 1 将决定如何 apoE4 会导致胰岛素抵抗。对 apoER2 相互作用蛋白的无偏见查询以及随后的研究培养的内皮细胞中的胰岛素转运表明 Dab2 相互作用蛋白 (DAB2IP) 和 JIP1，两者均调节 JNK 信号传导。在 apoE3- 和 apoE4- 中使用内皮细胞特异性缺失表达小鼠，我们将测试 apoE4 通过内皮 apoER2 促进胰岛素抵抗的假设通过 DAB2IP 和 JIP1 依赖机制。目标 2 将决定 apoE4 如何促进动脉粥样硬化和血栓形成。内皮细胞中与 apoER2 和 apoE4 相互作用的蛋白质研究内皮细胞 vWF 分泌和单核细胞粘附的增强表明可能参与蛋白磷酸酶 PP2A。在表达 apoE3 和 apoE4 的小鼠中，我们将检验 apoE4 的假设通过内皮 apoER2 相关的 PP2A 激活促进动脉粥样硬化和血栓形成。在目标 1 和 2 中，我们还将使用基于 GWAS 的策略来确定小鼠的操作基因是否是修饰基因影响人类 apoE4 相关代谢和血管疾病风险的基因。在目标 3 中，我们将设计针对 apoE4 的干预措施及其作用机制，以保持代谢和血管健康。在 apoE4 小鼠 JNK 抑制对胰岛素抵抗的影响以及 PP2A 抑制对胰岛素抵抗的影响将评估动脉粥样硬化和血栓形成。我们还将使用碱基编辑来对 apoE4 进行基因校正 apoE4小鼠肝细胞体内的apoE3。编辑效率及其对 apoE4 相关胰岛素的影响将评估抵抗力、动脉粥样硬化和血栓形成。通过实现这些目标并解开 apoE4如何促进心脏代谢疾病，预计可以开发出新的疗法来中和 15% 至 20% 人口中代谢和血管疾病的重要遗传因素。