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: 9929937
• 负责人: George A Truskey
• 金额: $ 5.71万
• 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/9929937
• 关键词: 20 year old; Aging; Alternative Splicing; Animal Model; Atherosclerosis; Biological Markers; Blood Vessels; Cardiovascular Diseases; Cause of Death; Cell Nucleus; Cells; Cessation of life; Clinical Research; Complement; Crystallization; Cultured Cells; Development; Disease; Disease Progression; Disease model; Effectiveness; Endothelial Cells; Endothelium; Environment; Exhibits; Gene Expression; Gene Mutation; Genes; Goals; Hour; Human Engineering; In Vitro; Individual; Inflammation Mediators; Inflammatory; Lamin Type A; Lamins; Measurement; Mediating; Nuclear Protein; Pathway interactions; Phenotype; Physiological; Point Mutation; Preparation; Process; Progeria; Rare Diseases; Signal Transduction; Smooth Muscle Myocytes; Structure; Syndrome; Testing; Tissue Engineering; Treatment Effectiveness; Vascular Endothelium; biomarker identification; cell type; drug candidate; human tissue; in vivo; induced pluripotent stem cell; insight; microphysiology system; mouse model; novel; novel therapeutics; parent grant; transcriptome sequencing

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 the overall 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. The goal of this diversity supplement is to support Ms. Crystal Kennedy to examine how the vascular endothelium is altered by HGPS and how the function of endothelial cells and smooth muscle cells is altered a gene editing mechanism to correct the disease. In Aim 1 of the Supplement we will use RNASeq to examine the differential change in gene expression of HGPS endothelial cells and compare with the results from healthy endothelium. This aim complements AIM 1.2 of the parent grant and will enable us to identify functional pathways altered by HGPS. In Aim 2, we will correct HGPS using gene editing and assess the impact on the function of TEBVs and endothelium. This study is consistent with Specific Aim 3 of the parent grant, which aims to assess novel HGPS treatments.

抽象的 哈钦森-吉尔福德早衰综合症 (HGPS) 是一种罕见的加速衰老的常染色体显性遗传疾病 导致 7 至 20 岁之间的死亡。该疾病源于点突变，产生 核蛋白核纤层蛋白 A（称为早老蛋白）的交替剪接形式，在细胞核中积累。 HGPS 小鼠模型与 HGPS 核纤层蛋白基因突变表现出许多表型相似性，但是 动脉粥样硬化不会发展，这表明动物模型的适用性有限。由于心血管 疾病是 HGPS 患者死亡的主要原因，我们建议使用一种新型组织 设计血管微生理系统来开发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。我们开发了动脉规模 使用来源于平滑肌细胞（SMC）的内皮化组织工程血管（TEBV） 使用健康细胞和 HGPS 细胞诱导多能干细胞 (iPSC)。 TEBV 可以生产并 在制备后几个小时内在生理流动条件下进行灌注，并表现出内皮介导的 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 提供了比塑料上培养的细胞更真实的体外环境，可以帮助推进 发现新疗法和识别生物标志物的过程。在整个项目中，我们将测试 组织工程血管由来自 HGPS 个体的细胞制成的假设 在体外重现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。这种多样性补充的目标是支持 Crystal Kennedy 女士检查 HGPS 如何改变血管内皮以及血管内皮的功能如何 内皮细胞和平滑肌细胞通过改变基因编辑机制来纠正疾病。目标 1 在补充材料中，我们将使用 RNASeq 来检查 HGPS 基因表达的差异变化 内皮细胞并与健康内皮细胞的结果进行比较。该目标是对 AIM 1.2 的补充 家长资助，将使我们能够识别 HGPS 改变的功能途径。在目标 2 中，我们将纠正 HGPS 使用基因编辑并评估对 TEBV 和内皮功能的影响。这项研究是 与母基金的具体目标 3 一致，该目标旨在评估新型 HGPS 治疗方法。