In Vitro Human Tissue-Engineered Blood Vessel Disease Model of Progeria

早衰症体外人体组织工程血管疾病模型

• 批准号: 9980460
• 负责人: George A Truskey
• 金额: $ 46.21万
• 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 2022-05-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9980460
• 关键词: 20 year old; Address; Aging; Alternative Splicing; Animal Model; Animals; Antisense Oligonucleotides; Apoptosis; Arterial Fatty Streak; Arteries; Atherosclerosis; Autophagocytosis; Bacterial Artificial Chromosomes; Biological Markers; Blood Vessels; CCI-779; Cardiovascular Diseases; Cardiovascular system; Cause of Death; Cell Aging; Cell Count; Cell Differentiation process; Cell Nucleus; Cells; Cellularity; Cessation of life; Clinical Research; Combined Modality Therapy; Coronary artery; Cultured Cells; Development; Disease; Disease Progression; Disease model; Drug Combinations; Effectiveness; Endothelium; Ensure; Environment; Epigenetic Process; Exhibits; Farnesyl Transferase Inhibitor; Fibroblasts; Fibrosis; Gene Expression; Gene Mutation; Genes; Hour; Human; Human Engineering; Impairment; In Vitro; Individual; Inflammation; Inflammation Mediators; Inflammatory; Knock-in; Lamin Type A; Lamins; Lesion; Lipids; Lonafarnib; Measurement; Medial; Mediating; Methylene blue; Mitochondria; Mitosis; Modeling; Mus; Myocardial Infarction; Nuclear; Nuclear Protein; Oxidative Stress; Parents; Patients; Perfusion; Pharmaceutical Preparations; Pharmacotherapy; Phenotype; Physiological; Point Mutation; Pravastatin; Preparation; Process; Production; Progeria; Proteins; RNA Splicing; Rare Diseases; Running; Rupture; Shapes; Signal Pathway; Signal Transduction; Site; Smooth Muscle Myocytes; Stroke; Structure; Syndrome; System; Testing; Therapeutic; Thick; Tissue Engineering; Treatment Effectiveness; Vascular Diseases; Zoledronic Acid; aged; biomarker identification; calcification; carbene; clinical candidate; drug action; drug candidate; histone methylation; human tissue; improved; in vivo; in vivo Model; induced pluripotent stem cell; microphysiology system; mouse model; normal aging; novel; novel therapeutics; osteogenic; protein degradation; response

Abstract Hutchinson-Gilford Progeria Syndrome (HGPS) is a rare autosomal dominant disease of accelerated aging which leads to death between 7 and 20 years of age. The disease arises from point mutations that produce an alternately spliced form of the nuclear protein lamin A, known as progerin, that accumulates in the cell nucleus. Mouse models of HGPS exhibit many phenotypical similarities with the HGPS lamin gene mutation, but atherosclerosis does not develop, suggesting a limit to the suitability of animal models. Since cardiovascular disease represents the primary cause of death among those with HGPS, we propose to use a novel tissue engineered blood vessel microphysiological system to develop biomarkers for the disease and assess the effectiveness of treatment against relevant physiological measurements. We have developed arteriolar-scale endothelialized tissue-engineered blood vessels (TEBVs) using smooth muscle cells (SMCs) derived from induced pluripotent stem cells (iPSCs) using healthy and HGPS cells. The TEBVs can be produced and perfused at physiological flow conditions within a few hours of preparation and exhibit endothelial-mediated vasoactivity and respond to inflammatory mediators. We can perform standard functional tests and examine the effects of inflammatory signals, thus tracking the progression of the disease in the same vessel. The HGPS-TEBVs provide a more realistic in vitro environment than cells cultured on plastic and can help advance the process of discovering novel therapeutics and identification of biomarkers. In this project, we will test the hypotheses that tissue-engineered blood vessels made with cells derived from individuals with HGPS recapitulate in vitro the structure and activity found in vivo and can aid in assessing the effectiveness and mode of action suitable drug candidates for clinical studies. In Aim 1, we will test the hypothesis that TEBVs with cells derived from HGPS patients have the same phenotype as a mouse model of HGPS. We will assess (1) the relative contribution of reduced cell number and oxidative stress on the altered function of HGPS- TEBVs, (2) the effect of flow on EC NRF2 activity and oxidative genes it regulates, and (3) compare TEBV structure and function with the mouse model for HGPS. Control conditions will consist of TEBVs prepared with cells derived from a parent of one of the HGPS patients. In Aim 2, we will modify our system to run multiple TEBVs simultaneously and test the hypothesis that combination therapies have been ineffective because they have not restored SMC number, differentiation, and vasoactivity. In Aim 3, we will assess the suitability of novel treatments for progeria to alter the HGPS phenotype in the TEBVs. We will examine the effect of agents which improve mitochondrial function and or protein degradation, alone or in combination with lonafarnib and anti-sense oligonucleotides that inhibit progerin production. Corresponding studies in mice will be performed to assess whether the HGPS-TEBV model reproduces changes to vessels found in mouse model of HGPS.

抽象的 Hutchinson-Gilford progeria综合征（HGP）是一种罕见的常染色体显性疾病的加速衰老疾病 导致7至20岁之间的死亡。该疾病来自产生的点突变 核蛋白层粘连蛋白A的剪接形式，称为雌蛋白，积聚在细胞核中。 HGPS的小鼠模型与HGPS层粘连蛋白基因突变表现出许多表型相似性，但是 动脉粥样硬化没有发展，这表明动物模型的适用性有限。由于心血管 疾病代表了HGP患者死亡的主要原因，我们建议使用新的组织 工程的血管微生物生理系统开发了该疾病的生物标志物，并评估 针对相关生理测量的治疗有效性。我们已经开发了小动脉尺度 使用源自源自的平滑肌细胞（SMC）的内皮化组织工程血管（TEBV） 使用健康和HGPS细胞诱导多能干细胞（IPSC）。可以生产tebvs 在制备后的几个小时内，在生理流动条件下灌注并展示了内皮介导的 血管活性和对炎症介质的反应。我们可以执行标准功能测试并检查 炎症信号的影响，从而跟踪同一血管中疾病的进展。这 HGPS-TEBV与在塑料上培养的细胞相比，在体外环境中提供了更现实的体外环境 发现新型疗法和生物标志物鉴定的过程。在这个项目中，我们将测试 假设由源自HGP的个体衍生的细胞制成的组织设计的血管 在体外概括体内发现的结构和活性，可以帮助评估有效性和 作用方式适合用于临床研究的候选药物。在AIM 1中，我们将测试TEBVS的假设 来自HGPS患者的细胞具有与HGP小鼠模型相同的表型。我们将评估 （1）细胞数减少和氧化应激对HGPS-功能改变的相对贡献 TEBV，（2）流动对IT调节的EC NRF2活性和氧化基因的影响，（3）比较TEBV HGP的鼠标模型的结构和功能。控制条件将由准备的TEBV组成 源自HGPS患者之一的母体的细胞。在AIM 2中，我们将修改系统以运行多个 同时进行TEBV并检验组合疗法无效的假设，因为它们 尚未恢复SMC数，分化和血管活性。在AIM 3中，我们将评估 后代的新型治疗方法改变了TEBV中的HGP表型。我们将检查代理的效果 改善线粒体功能和 /或蛋白质降解，单独或与lonafarnib和 抑制孕激素产生的抗义寡核苷酸。将对小鼠进行相应的研究 评估HGPS-TEBV模型是否会重现HGP小鼠模型中的血管的变化。