Project 1. Diabetes, triglyceride-rich lipoproteins, and advanced atherosclerosis

项目1. 糖尿病、富含甘油三酯的脂蛋白和晚期动脉粥样硬化

• 批准号: 10450861
• 负责人: Karin E. Bornfeldt
• 金额: $ 40.47万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-15 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10450861
• 关键词: ANGPTL3 gene; Address; Affect; Antisense Oligonucleotides; Apolipoproteins; Arterial Fatty Streak; Atherosclerosis; CREB1 gene; Cardiovascular Diseases; Caspase; Cessation of life; Cholesterol; Complement; Cyclic AMP-Responsive DNA-Binding Protein; Data; Diabetes Mellitus; Diabetic mouse; Event; Exhibits; Health; Hemorrhage; Hepatic; Human; Human Genetics; Hypertriglyceridemia; Inflammasome; Insulin Receptor; Insulin Resistance; Insulin deficiency; Insulin-Dependent Diabetes Mellitus; LDL Cholesterol Lipoproteins; Lesion; Lesion by Morphology; Link; Lipids; Lipoproteins; Mediating; Metabolic syndrome; Metabolism; Modeling; Mus; Mutation; Myeloid Cells; Necrosis; Non-Insulin-Dependent Diabetes Mellitus; Pathway interactions; Patients; Phenotype; Plasma; Prediabetes syndrome; Prevention strategy; Proteins; Publishing; Receptor Signaling; Residual state; Risk Factors; Role; Serum; Severities; Testing; Time; Triglyceride Metabolism; Triglycerides; atherosclerosis risk; cardiovascular disorder risk; diabetic; diabetic patient; in vivo; insight; interest; loss of function mutation; macrophage; mouse model; non-diabetic; novel; novel strategies; particle; prevent; trait; transcription factor; treatment effect; treatment strategy; uptake

Both type 1 diabetes mellitus (T1DM), type 2 diabetes mellitus (T2DM) and metabolic syndrome (MetS; also known as pre-diabetes) increase the risk of atherosclerosis, the leading cause of cardiovascular disease (CVD) events and death in diabetic patients. Even with statin treatment to effectively lower LDL cholesterol, diabetes is associated with increased CVD risk linked to abnormal metabolism of triglyceride-rich lipoproteins (TRLs) and their remnant lipoprotein particles (RLPs). Novel strategies to enhance TRL clearance have therefore recently generated significant interest, especially in the setting of diabetes. This application uses three strategies to increase TRL and RLP clearance in diabetes; i) inhibition of apolipoprotein C3 (APOC3), ii) inhibition of angiopoietin-like 3 (ANGPTL3), and iii) hepatic expression of an active form of the transcription factor CREB-H (cAMP response element-binding protein H). The proposed studies build on strong preliminary data demonstrating that diabetes accelerates progression of advanced lesions and that normalizing circulating levels of TRLs prevents atherosclerosis in diabetic mice. Furthermore, diabetes causes increased macrophage death associated with accumulation of cellular cholesterol and caspase 11 activation, which we hypothesize contributes to the more advanced atherosclerotic lesion phenotype. Collectively, our observations support the following hypothesis: Diabetes accelerates advanced atherosclerosis by increasing APOC3 and associated TRLs and RLPs, which in turn induce macrophage lipid loading and exacerbate the death of macrophages by pyroptosis. Such a mechanism could help explain the increased CVD risk in subjects with diabetes. The first specific aim will address whether APOC3-enriched TRLs/RLPs are required for diabetes- mediated atherosclerosis progression. We propose to test this hypothesis and to clarify the role of APOC3 by normalizing plasma APOC3 and/or TRL/RLP levels in mouse models of T1DM and MetS/T2DM by three distinct approaches; the use of an Apoc3 antisense oligonucleotide (ASO), an Angptl3 ASO, which reduces TRLs and other lipoproteins independent of hepatic Apoc3 expression, and hepatic expression of CREB-H, which lowers TRLs without affecting total plasma levels of APOC3. Contrasting the effects of these treatments will provide new insights into the role of APOC3 versus TRL/RLP lowering. The second specific aim will determine whether pyroptosis mediated by APOC3-enriched TRLs/RLPs contributes to lesional macrophage death and advanced lesions in diabetes. We propose to mechanistically test the importance of this pathway by using mice deficient in caspase 11 and gasdermin D, its downstream effector, in myeloid cells. We also expect that pyroptosis will be prevented by TRL-lowering in diabetic mice. The in vivo studies will be complemented by mechanistic studies in macrophages to clarify the pathways responsible in diabetes-induced macrophage death, and by studies addressing mechanisms whereby reduced insulin receptor signalling leads to increased levels of APOC3 in models of T1DM and MetS/T2DM.

两种类型1糖尿病（T1DM），2型糖尿病（T2DM）和代谢综合征（Mets；也 被称为糖尿病前）增加了动脉粥样硬化的风险，动脉粥样硬化是心血管疾病（CVD）的主要原因 糖尿病患者的事件和死亡。即使汀类药物治疗可有效降低LDL胆固醇，糖尿病 与与富含甘油三酸酯的脂蛋白（TRL）异常代谢有关的CVD风险增加有关 以及它们的残留脂蛋白颗粒（RLP）。因此，增强TRL许可的新型策略具有 最近引起了重大兴趣，尤其是在糖尿病的情况下。此应用程序使用三个 增加糖尿病中TRL和RLP清除率的策略； i）抑制载脂蛋白C3（APOC3），II） 抑制血管生成素样3（Angptl3）和iii）肝表达转录的活性形式 因子CREB-H（CAMP响应元件结合蛋白H）。拟议的研究以强大的初步为基础 数据表明，糖尿病会加速晚期病变的进展并使循环正常 TRL的水平阻止了糖尿病小鼠的动脉粥样硬化。此外，糖尿病会导致巨噬细胞增加 与细胞胆固醇和caspase 11激活相关的死亡，我们假设 有助于更先进的动脉粥样硬化病变表型。总的来说，我们的观察结果支持 以下假设：糖尿病通过增加apoc3和 相关的TRL和RLP，这又诱导巨噬细胞脂质载荷并加剧了死亡 巨噬细胞通过凋亡。这样的机制可以帮助解释患者的CVD风险增加 糖尿病。第一个具体目的将解决糖尿病是否需要APOC3富集的TRL/RLP- 介导的动脉粥样硬化进展。我们建议检验这一假设，并阐明Apoc3的作用 将T1DM和MetS/T2DM鼠标模型中的等离子体APOC3和/或TRL/RLP归一化 不同的方法；使用APOC3反义寡核苷酸（ASO），ANGPTL3 ASO，可减少 TRL和其他脂蛋白独立于肝APOC3表达和CREB-H的肝表达， 降低TRL而不会影响apoc3的总血浆水平。对比这些治疗的影响 将提供有关APOC3与TRL/RLP降低的作用的新见解。第二个特定目标将 确定富含APOC3的TRL/RLP介导的凋亡是否有助于病变巨噬细胞 糖尿病的死亡和晚期病变。我们建议通过机械学测试该途径的重要性 使用缺乏caspase 11的小鼠和髓样细胞中的下游效应子Gasdermin D。我们也期望 糖尿病小鼠中的TRL降低将预防凋亡。体内研究将由 巨噬细胞的机械研究以阐明糖尿病诱导的巨噬细胞的途径 死亡，以及通过解决机制的研究，从而减少胰岛素受体信号传导导致增加 T1DM和MetS/T2DM模型中的APOC3级别。