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: 9759965
• 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 2021-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9759965
• 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; Drug effect disorder; 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 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.

摘要 Hutchinson-Gilford Progeria综合征(HGPS)是一种罕见的加速衰老的常染色体显性遗传病 这会导致7到20岁的儿童死亡。这种疾病是由点突变引起的 另一种剪接形式的核蛋白层蛋白A，称为孕激素，在细胞核中积累。 HGPS小鼠模型显示出许多与HGPS lamin基因突变的表型相似之处，但 动脉粥样硬化不会发展，这表明动物模型的适用性受到限制。自心血管疾病以来 疾病是HGPS患者的主要死亡原因，我们建议使用一种新的组织 工程血管微生理系统，为疾病开发生物标记物并评估 对照相关生理指标的治疗效果。我们已经开发出微动脉刻度 血管内皮化组织工程血管(TEBV)的构建 使用健康和HGPS细胞诱导多能干细胞(IPSCs)。TEBV可以生产和 在生理流动条件下准备几小时内灌流，并显示内皮细胞介导的 血管活性和对炎症介质的反应。我们可以执行标准的功能测试并检查 炎症信号的影响，从而在同一血管中跟踪疾病的进展。这个 HGPS-TEBV提供了比塑料上培养的细胞更逼真的体外环境，并有助于 发现新疗法和鉴定生物标记物的过程。在这个项目中，我们将测试 假设组织工程血管是由来自HGPS患者的细胞制成的 在体外概括在体内发现的结构和活性，有助于评估有效性和 作用方式适合临床研究的候选药物。在目标1中，我们将检验TEBV的假设 对于来自HGPS的细胞，患者具有与HGPS小鼠模型相同的表型。我们将评估 (1)细胞数量减少和氧化应激对HGPS功能改变的相对贡献 TEBV；(2)流动对EC NRF2活性及其调控基因的影响；(3)TEBV的比较 用于HGPS的鼠标模型的结构和功能。控制条件将由准备的TEBV组成 来自其中一名HGPS患者父母的细胞。在目标2中，我们将修改我们的系统以运行多个 TEBV同时测试联合疗法无效的假设，因为它们 尚未恢复SMC数量、分化和血管活性。在目标3中，我们将评估 早衰症的新治疗方法改变TEBV中的HGPS表型。我们将检查代理人的效果 可改善线粒体功能和/或蛋白质降解，单独或与氯那法尼和 抑制孕激素产生的反义寡核苷酸。将在老鼠身上进行相应的研究，以 评估HGPS-TEBV模型是否再现了HGPS小鼠模型中发现的血管的变化。