Systemic Inflammation in Microphysiological Models of Muscle and Vascular Disease

肌肉和血管疾病微生理模型中的全身炎症

• 批准号: 10013428
• 负责人: George A Truskey
• 金额: $ 25.09万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-01 至 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10013428
• 关键词: Affect; Animal Model; Anti-inflammatory; Antibodies; Arteries; Atherosclerosis; Attenuated; Autoimmune Diseases; Biological Markers; Biological Models; Biomechanics; Blood Vessels; CRISPR/Cas technology; Cardiac; Cardiovascular Diseases; Cell Proliferation; Cells; Cholesterol; Chronic; Chronic Disease; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Development; Disease; Disease Progression; Disease model; Drug Modelings; Effectiveness; Endothelial Cells; Endothelium; Environment; Event; Exercise; Exposure to; Feedback; Foam Cells; Functional disorder; Gene Expression; Genes; Genetic Variation; Goals; Heart; Human; Immune; Immune response; In Vitro; Individual; Inflammation; Inflammatory; Inflammatory Response; Interleukin 6 Receptor; Interleukin-6; Laboratories; Lipid-Laden Macrophage; Lipids; Lipoproteins; Liquid substance; Low-Density Lipoproteins; Mediating; Modeling; Muscle; Muscle function; Muscular Atrophy; Mutation; Myocardium; Myopathy; Pathology; Pattern; Perfusion; Pharmaceutical Preparations; Pharmacotherapy; Phase; Phenotype; Population; Process; Proprotein Convertases; Rheumatoid Arthritis; Risk; Role; Severities; Severity of illness; Single Nucleotide Polymorphism; Site; Skeletal Muscle; Smooth Muscle Myocytes; Structure; Subtilisins; Symptoms; System; Technology; Testing; Therapeutic Intervention; Therapeutic exercise; Tissue Engineering; Tissues; Variant; Vascular Diseases; base; cardiovascular disorder risk; clinically relevant; cytokine; disease phenotype; drug development; drug discovery; drug testing; endothelial dysfunction; gain of function mutation; genetic variant; human disease; improved; in vitro Model; in vivo; induced pluripotent stem cell; joint inflammation; loss of function; loss of function mutation; macrophage; microphysiology system; novel therapeutics; population based; protein profiling; repaired; response; shear stress; spatial temporal variation; therapeutic target

ABSTRACT The initiation and progression of atherosclerosis is influenced by systemic inflammation and individuals suffering from autoimmune diseases, such as rheumatoid arthritis, have increased risk of developing cardiovascular diseases. Likewise, chronic and systemic inflammation in rheumatoid arthritis induces muscle wasting and loss of function. Therapies that reduce inflammation effectively treat rheumatoid arthritis and have the potential to reduce the severity of cardiovascular disease. To overcome limitations of animal models replicating some key disease phenotypes, but not the underlying mechanisms, we established functional human microphysiological systems (hMPS) for healthy human skeletal and cardiac muscle and endothelialized tissue-engineered blood vessels (eTBEVs) using primary and iPS-derived cells and assessed the response to drugs and pro-inflammatory cytokines. These models replicate the structure and key functions of the native tissue and maintain their structure and function for at least 4 weeks. These in vitro tissue systems accurately model the response to drugs. Our goal in this project is to develop clinically relevant hMPS disease models to examine rheumatoid arthritis (RA) risk for muscle dysfunction and atherosclerosis and the role of exercise in attenuating disease-associated inflammation. To meet this goal, we will expand our preliminary results to develop and validate an early atherosclerosis model that uses flow conditions promoting endothelial dysfunction, macrophage accumulation, foam cell formation, and altered vasoactivity. We will reproduce the RA phenotype in skeletal and cardiac muscle through addition of macrophages and cytokines present in RA, and demonstrate that simulated exercise conditions on muscle produce myokines that reduce inflammation in this RA model. Then, we will develop an integrated perfusion system for eTEBVs, skeletal and cardiac muscle and show that the RA model can increase macrophage accumulation in eTEBVs and cardiac bundles, and assess the response to exercise and drugs to treat atherosclerosis and inflammation. We will use CRISPR gene editing technology to generate mutations to proprotein convertase subtilisin/kexin type 9 (PCSK9) and genes that affect IL-6 shedding to assess their impact on endothelial dysfunction and foam cell formation in eTEBVs, and inflammation in skeletal and cardiac muscle bundles. We will profile cytokines and metabolites in the models with and without RA, and demonstrate that disease progression and biomarkers are reduced in the presence of common anti-inflammatory therapeutic interventions for atherosclerosis, and assess the effect of exercise. Likewise, in the RA muscle model, we will examine whether gene variants produce alterations in cytokine profiles impacting muscle function and response to exercise; these may point toward new disease-associated biomarkers and therapeutic targets. Results of this project will provide a general framework for in vitro modeling of atherosclerosis and autoimmune diseases and the role of gene variants in disease severity and drug development.

摘要 动脉粥样硬化的发生和发展受全身炎症和个体的影响。 患有自身免疫性疾病，如类风湿性关节炎，会增加发病的风险 心血管疾病。类似地，类风湿性关节炎的慢性和全身性炎症导致肌肉 虚弱和丧失功能。有效消炎的疗法治疗类风湿性关节炎和 有可能降低心血管疾病的严重程度。克服动物的局限性 复制了一些关键疾病表型的模型，但没有复制潜在的机制，我们建立了 用于健康人体骨骼肌和心肌的功能性人体微生理系统(HMPS) 使用原代和iPS来源细胞的内皮化组织工程血管(ETBEV)及其评估 对药物和促炎细胞因子的反应。这些模型复制了结构和关键功能 并保持其结构和功能至少4周。这些体外组织 系统准确地模拟了对药物的反应。我们在这个项目中的目标是开发与临床相关的 用于检测类风湿关节炎(RA)肌肉功能障碍和动脉粥样硬化风险的HMPS疾病模型 以及运动在减轻疾病相关炎症方面的作用。为了实现这一目标，我们将扩大我们的 开发和验证使用血流条件促进的早期动脉粥样硬化模型的初步结果 内皮功能障碍、巨噬细胞聚集、泡沫细胞形成和血管活性改变。我们会 通过添加巨噬细胞和细胞因子在骨骼肌和心肌中复制RA表型 存在于类风湿关节炎中，并证明了肌肉上的模拟运动条件产生的肌动蛋白可以减少 这个RA模型中的炎症。然后，我们将为eTEBV，骨骼开发一个集成的灌流系统 和心肌，表明RA模型可以增加eTEBV和ETEBV的巨噬细胞聚集 心束，评估运动和治疗动脉粥样硬化和炎症的药物的反应。 我们将使用CRISPR基因编辑技术产生突变，以产生枯草杆菌/可信原蛋白转换酶 9型(PCSK9)和影响IL-6脱落的基因，以评估它们对内皮功能障碍和 ETEBV中泡沫细胞的形成，以及骨骼肌束和心肌束中的炎症。我们将为您介绍 细胞因子和代谢物在有和没有类风湿关节炎的模型中的变化，并表明疾病的进展和 在存在常见的抗炎治疗干预措施的情况下，生物标志物会减少 动脉粥样硬化，并评估运动的效果。同样，在RA肌肉模型中，我们将检查 基因变异会导致影响肌肉功能和运动反应的细胞因子谱发生变化； 这些可能指向新的疾病相关生物标记物和治疗靶点。本项目的成果 将为动脉粥样硬化和自身免疫性疾病的体外建模提供一个通用的框架，以及 基因变异在疾病严重性和药物开发中的作用。