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

基本信息

批准号：
10471015
负责人：
George A Truskey
金额：
$ 7.33万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
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/10471015
关键词：
Affect Animal Model Anti-Inflammatory Agents 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 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 Proteins 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 effectiveness 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 progression marker repaired response shear stress spatial temporal variation systemic inflammatory response therapeutic target

项目摘要

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.

抽象的动脉粥样硬化的开始和进展受系统性炎症和个体的影响患有自身免疫性疾病（例如类风湿关节炎）患有发展的风险心血管疾病。同样，类风湿关节炎中的慢性和全身性炎症会诱导肌肉浪费和功能丧失。减少炎症的疗法有效治疗类风湿关节炎和有可能减少心血管疾病的严重程度。克服动物的局限性复制某些关键疾病表型的模型，但不是基本机制，我们建立了健康人骨骼肌和心脏肌肉的功能性人类微生物生理系统（HMP）使用原始和IPS衍生的细胞进行内皮化组织工程血管（ETBEV），并评估对药物和促炎性细胞因子的反应。这些模型复制结构和关键功能天然组织的结构和功能至少4周。这些体外组织系统准确地模拟对药物的反应。我们在这个项目中的目标是开发临床相关 HMPS病模型检查类风湿关节炎（RA）肌肉功能障碍和动脉粥样硬化的风险运动在衰减与疾病相关的炎症中的作用。为了实现这一目标，我们将扩大我们的初步结果以开发和验证使用促进流条件的早期动脉粥样硬化模型内皮功能障碍，巨噬细胞积累，泡沫细胞的形成和血管活性改变。我们将通过添加巨噬细胞和细胞因子来重现骨骼和心肌中的RA表型在RA中存在，并证明对肌肉的模拟运动条件会产生肌动物，从而减少在此RA模型中的炎症。然后，我们将开发一个用于ETEBV的集成灌注系统和心肌，并表明RA模型可以增加ETEBV中的巨噬细胞积累心脏束，并评估对运动和药物治疗动脉粥样硬化和炎症的反应。我们将使用CRISPR基因编辑技术来生成突变以促蛋白转化酶枯草蛋白/kexin 9型（PCSK9）和影响IL-6脱落的基因，以评估其对内皮功能障碍的影响和 ETEBV中的泡沫细胞形成，以及骨骼和心脏肌肉束的炎症。我们将概括有或没有RA的模型中的细胞因子和代谢产物，并证明了疾病进展和在存在常见的抗炎治疗干预措施的情况下，生物标志物被降低动脉粥样硬化并评估运动的影响。同样，在RA肌肉模型中，我们将检查是否是否基因变异会导致影响肌肉功能和对运动反应的细胞因子谱的改变；这些可能指向新的与疾病相关的生物标志物和治疗靶标。这个项目的结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一般框架，基因变异在疾病严重程度和药物发育中的作用。