Synthetic 3D Model of the Carotid Artery to Study Exercise-Induced Changes in Endothelial Gene Expression

用于研究运动引起的内皮基因表达变化的颈动脉合成 3D 模型

• 批准号: 10406351
• 负责人: Alvaro N Gurovich
• 金额: $ 15.1万
• 依托单位: UNIVERSITY OF TEXAS EL PASO
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-05-17 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10406351
• 关键词: 3-Dimensional; 3D Print; Actins; Aerobic Exercise; Affect; Age; Anatomy; Applications Grants; Area; Atherosclerosis; Biochemical; Biocompatible Materials; Biological Availability; Biomechanics; Blood Vessels; Blood flow; Cardiovascular Diseases; Cardiovascular system; Carotid Arteries; Cells; Characteristics; Clinical; Clinical Markers; Collagen; Computer Models; Cultured Cells; Data; Digital Imaging and Communications in Medicine; Down-Regulation; Endothelial Cells; Endothelin-1; Endothelium; Epigenetic Process; Epoprostenol; Exercise; Future; Gene Expression; Genes; Genetic; Genetic Transcription; Glycocalyx; Health Professional; Homeostasis; Human; Hypertension; Image; Immunohistochemistry; In Vitro; Inflammatory; Intercellular adhesion molecule 1; Laboratories; Length; Magnetic Resonance Imaging; Mechanics; Medicine; Messenger RNA; Modeling; Molecular; Nitric Oxide; Nurses; Obesity; Oxidative Stress; Pathway interactions; Patients; Pattern; Physiological; Polymerase Chain Reaction; Printing; Process; Property; Proteins; Rehabilitation therapy; Reporting; Research; Rest; Reverse Transcription; Shapes; Stress; Stress Fibers; Stroke; Structural Protein; Surface; Technology; Testing; Translations; Tunica Intima; Vascular Cell Adhesion Molecule-1; Western Blotting; alpha Actinin; base; bioprinting; cardiovascular health; cardiovascular risk factor; cerebrovascular; endothelial dysfunction; exercise intensity; exercise physiologist; exercise program; experimental study; hemodynamics; heparin proteoglycan; human model; hypercholesterolemia; imaging study; improved; in vivo; mechanotransduction; personalized medicine; physical therapist; programs; protective factors; protein expression; receptor; shear stress; standard care; stressor; stroke survivor; three-dimensional modeling; transcription factor; translational approach

Summary Approximately nine out of 10 cerebrovascular attacks are due to atherosclerosis. Additionally, endothelial dysfunction is currently accepted as the first pathophysiological step toward atherosclerosis. Endothelial cell homeostasis and gene expression is highly regulated via shear stress, which is directly associated with blood flow changes. Aerobic exercise (AX) has been associated with improved cardiovascular (CV) health. However, only ~50% of the beneficial effects of AX are explained via improvements on traditional CV risk factors (e.g., hypertension, hypercholesterolemia, obesity). The remainder ~50% of the beneficial effects of AX are unknown. Moreover, traditionally controlled AX does not provide personalized medicine, which could account for a high number of non-responders. Therefore, the main purpose of this proposal is to develop a 3D synthetic model of the human carotid artery using 3D bio-printing technology to simulate in vivo personalized AX-induced blood flow patterns and endothelial shear stress and to determine gene expression/transcription and molecular changes in endothelial cultured cells in vitro. Based on previous reports and our preliminary data we hypothesize that a 3D synthetic model of the carotid artery will respond to exercise-induced blood flow patterns as a normal carotid artery. In addition, we hypothesize that endothelial cultured cells under similar blood flow patterns and shear stress will increase the expression of atherosclerosis-protective mRNA/proteins (e.g., eNOS, PGI2, and SOD) and structural mRNA/proteins (e.g., actin, heparin sulfate proteoglycan [glycocalyx], and α-actinin-bundled stress fibers), and a decrease of pro- atherosclerosis and pro-inflammatory mRNA/protein expression (e.g., ICAM-1, VCAM-1, and ET-1) in a similar intensity-dependent manner. First, we will determine biomechanical properties (e.g., vessel distensibility and compliance) of the carotid artery in vivo during resting conditions and at 3 AX intensities in healthy, young subjects, patients with stroke, and age-matched controls. Then, subjects will undergo a magnetic resonance imaging (MRI) study to determine the exact shape (e.g., length and contour) of same tested carotid artery and the images will be used to build a 3D synthetic model via 3D bio-printing. The 3D synthetic model will mimic more anatomical and hemodynamic conditions, which will allow for more physiological in vitro experiments. Secondly, we will perform several flow patterns in endothelial cultured cells seeded on the 3D synthetic model. Flow patterns will be similar to those patterns observed during in vivo studies. After applying the different flow patterns, cells will be collected and processed to determine changes in specific gene transcription factors and protein expression. By characterizing blood flow patterns during different intensities of AX and determining the gene expression/transcription and molecular changes in endothelial cells under these same blood flow patterns, using a ‘reverse’ translation approach, we will explain the mechanisms of the cardiovascular protective factors associated with AX as personalized medicine, especially in stroke survivors.

概括 大约十分之九的脑血管发作是由于动脉粥样硬化引起的。 目前认为功能障碍是动脉粥样硬化的第一个病理生理步骤。 体内平衡和基因表达通过剪切应力受到高度调节，这与血液直接相关 有氧运动 (AX) 与改善心血管 (CV) 健康有关。 只有约 50% 的 AX 有益效果可以通过改善传统的 CV 风险因素（例如， AX 的其余约 50% 的有益作用是 此外，传统控制的 AX 不提供个性化医疗，这可以说明。 因此，该提案的主要目的是开发 3D 方案。 利用3D生物打印技术模拟活体人体颈动脉合成模型 个性化 AX 诱导的血流模式和内皮剪切应力并确定基因 体外培养的内皮细胞的表达/转录和分子变化 基于之前的研究。 报告和我们的初步数据，我们打乱颈动脉的 3D 合成模型将做出反应 此外，我们将运动引起的血流模式视为正常的颈动脉。 在相似的血流模式和剪切应力下培养的细胞会增加 动脉粥样硬化保护性 mRNA/蛋白（例如 eNOS、PGI2 和 SOD）和结构 mRNA/蛋白（例如 肌动蛋白、硫酸肝素蛋白聚糖 [糖萼] 和 α-肌动蛋白捆绑的应力纤维），以及亲- 动脉粥样硬化和促炎性 mRNA/蛋白表达（例如 ICAM-1、VCAM-1 和 ET-1）具有相似的作用 首先，我们将确定生物力学特性（例如，血管扩张性和 在健康、年轻的人中，在静息条件下和 3 AX 强度下体内颈动脉的顺应性 然后，受试者将接受磁共振检查。 成像（MRI）研究以确定相同测试颈动脉的确切形状（例如长度和轮廓）和 这些图像将用于通过 3D 生物打印构建 3D 合成模型。 3D 合成模型将进行模仿。 更多的解剖学和血流动力学条件，这将允许进行更多的生理体外实验。 其次，我们将在 3D 合成模型上接种的内皮培养细胞中执行几种流动模式。 流动模式将与应用不同流动后在体内研究中观察到的模式相似。 模式，将收集和处理细胞以确定特定基因转录因子和 通过表征不同 AX 强度期间的血流模式并确定 在相同血流下内皮细胞的基因表达/转录和分子变化 模式，使用“反向”翻译方法，我们将解释心血管的机制 与 AX 作为个性化医疗相关的保护因素，尤其是中风幸存者。