Exosomes in the Pathogenesis of Diabetic Atherosclerosis & its Treatment Opportunities

外泌体在糖尿病动脉粥样硬化发病机制中的作用

• 批准号: 10266066
• 负责人: Robert Raffai
• 金额: --
• 依托单位: VETERANS AFFAIRS MED CTR SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10266066
• 关键词: Anti-Inflammatory Agents; Antiinflammatory Effect; Aorta; Applications Grants; Arterial Fatty Streak; Arteries; Atherosclerosis; Automobile Driving; Biomedical Engineering; Blood Circulation; Blood Glucose; CRISPR/Cas technology; Cardiovascular Diseases; Cell Culture Techniques; Cells; Cessation of life; Cholesterol; Cultured Cells; Data; Development; Diabetes Mellitus; Diabetic mouse; Diagnosis; Disease; Engineering; Enrollment; Exposure to; Gene Expression; General Population; Genes; Glucose; Goals; Healthcare Systems; Hematopoiesis; Hematopoietic stem cells; Human; Hyperglycemia; Immune; Immune system; In Vitro; Individual; Inflammation; Inflammatory; Infusion procedures; Interleukin-4; Leukocytes; Leukocytosis; Link; Lipid Bilayers; Lipid-Laden Macrophage; Lipids; Mediator of activation protein; Medical; Medical center; Metabolic; MicroRNAs; Modeling; Mus; Nucleic Acids; Pathogenesis; Peripheral; Peripheral arterial disease; Plasma; Process; Production; Property; Proteins; Reporting; Research; Risk; Role; Signal Transduction; Signaling Molecule; Source; Spleen; System; Testing; Therapeutic; Time; Veterans; base; cell type; cohort; cytokine; design; diabetic; disability; disorder control; exosome; extracellular vesicles; human subject; intercellular communication; macrophage; microvesicles; military veteran; monocyte; non-diabetic; novel; novel therapeutic intervention; oxidized low density lipoprotein; premature; protective factors; stem cell proliferation; stressor; systemic inflammatory response; trafficking; transcriptome sequencing; vascular inflammation; vesicular release

The goal of this proposal is to understand why people with diabetes develop severe fatty-rich plaques called atherosclerosis in arteries. Accelerated atherosclerosis has been linked to several types of cardiovascular diseases that cause disabilities and premature death amongst diabetic individuals. Unfortunately, such serious medical problems are very frequent among veterans enrolled in the VHA Health Care System as they are almost three times more likely to develop diabetes than people in the general population. One reason that has been identified to enhance atherosclerosis among diabetic individuals is high blood sugar, also called “hyperglycemia”. But exactly how hyperglycemia enhances atherosclerosis is not known. Our recent research focus has centered on exploring the role of microvesicles released by cells, called “exosomes”, as a source of inflammation in atherosclerosis. Preliminary findings presented in our revised grant proposal show that human diabetic subjects diagnosed with advance peripheral atherosclerotic cardiovascular disease accumulate pro-inflammatory exosomes in their bloodstream. Our findings also show that diabetic mice accumulate pro-inflammatory exosomes in their bloodstream. Furthermore, our findings show that such diabetic plasma exosomes can increase the number of white blood cells that accumulate in arteries when they are infused into non-diabetic mice. Interestingly, our data demonstrate that pro-inflammatory exosomes can be produced by macrophages cultured in glucose-rich medium that simulates diabetic hyperglycemia. Remarkably, our data also show that macrophages can produce anti-inflammatory exosomes when they cultured in medium that contains protective cytokines such as interleukin-4. Based on our extensive new findings, we propose to explore whether diabetic hyperglycemia enhances the progression of atherosclerosis through exosomes that communicate pro-inflammatory signaling in the immune system and the vessel wall. We also aim to produce exosomes that can serve to overcome the effects of hyperglycemia to suppress the progression of diabetic atherosclerosis. In our First Aim we will define the extent to which hyperglycemia causes the production of proinflammatory exosomes in the bloodstream of veterans suffering from advanced atherosclerotic disease. We will do the same by studying mouse models of diabetes. Next, we will test the ability of diabetic plasma exosomes to enhance vascular inflammation and atherosclerosis when infused into hyperlipidemic mice. We will also test whether macrophages cultured in medium that contains elevated levels of glucose or bad cholesterol called oxLDL, will produce exosomes that can induce atherosclerosis when infused into non-diabetic mice. Lastly, we will explore if a class of signaling molecule called microRNA carried by exosomes are responsible for their inflammatory signaling. In our Second Aim we will seek to produce therapeutic exosomes. Our strategy will consist of isolating exosomes from macrophages cultured in medium that contains protective factors including interleukin-4. Our second strategy will be to produce bioengineered exosomes from cultured macrophage tailored to produce desired levels of protective miRNA and test whether such exosomes can exert anti-inflammatory properties to reduce the progression of atherosclerosis in diabetic mice.

该提案的目的是了解为什么糖尿病患者会出现严重的富含脂肪的斑块，称为 动脉粥样硬化。加速动脉粥样硬化与多种类型的心血管疾病有关 导致糖尿病患者残疾和过早死亡的疾病。不幸的是，如此严重 在参加 VHA 医疗保健系统的退伍军人中，医疗问题非常频繁，因为他们几乎 患糖尿病的可能性是普通人群的三倍。原因之一是 已确定会加剧糖尿病患者动脉粥样硬化的是高血糖，也称为“高血糖”。 但高血糖究竟如何促进动脉粥样硬化尚不清楚。 我们最近的研究重点集中在探索细胞释放的微泡的作用，称为 “外泌体”，作为动脉粥样硬化炎症的来源。我们修订后的拨款中提出的初步调查结果 提案表明，人类糖尿病受试者被诊断患有晚期外周动脉粥样硬化性心血管疾病 疾病会在血液中积累促炎性外泌体。我们的研究结果还表明，糖尿病小鼠 在血液中积累促炎性外泌体。此外，我们的研究结果表明，此类糖尿病患者 血浆外泌体输注后可以增加动脉中积聚的白细胞数量 进入非糖尿病小鼠。有趣的是，我们的数据表明促炎外泌体可以通过 在模拟糖尿病高血糖的富含葡萄糖的培养基中培养的巨噬细胞。值得注意的是，我们的数据还 表明巨噬细胞在含有以下成分的培养基中培养时可以产生抗炎外泌体 保护性细胞因子，例如白介素-4。 基于我们广泛的新发现，我们建议探索糖尿病高血糖是否会增强 通过在免疫系统中传递促炎信号的外泌体来促进动脉粥样硬化的进展 系统和容器壁。我们还旨在生产可以克服以下影响的外泌体 高血糖可抑制糖尿病动脉粥样硬化的进展。 在我们的首要目标中，我们将定义高血糖导致促炎物质产生的程度 患有晚期动脉粥样硬化疾病的退伍军人血液中的外泌体。我们也会做同样的事 通过研究糖尿病小鼠模型。接下来，我们将测试糖尿病血浆外泌体增强糖尿病血浆的能力 当输注到高脂血症小鼠体内时，会引起血管炎症和动脉粥样硬化。我们还将测试是否 在含有升高水平的葡萄糖或称为 oxLDL 的坏胆固醇的培养基中培养的巨噬细胞，将 当注入非糖尿病小鼠体内时，会产生可诱导动脉粥样硬化的外泌体。最后，我们将探讨 外泌体携带的一类名为 microRNA 的信号分子是否对其炎症负责 发信号。 在我们的第二个目标中，我们将寻求生产治疗性外泌体。我们的策略将包括隔离 来自在含有白细胞介素 4 等保护因子的培养基中培养的巨噬细胞的外泌体。我们的 第二个策略是从培养的巨噬细胞中生产生物工程外泌体，以生产 所需水平的保护性 miRNA 并测试此类外泌体是否可以发挥抗炎特性 减少糖尿病小鼠动脉粥样硬化的进展。