Circulatory system and integrated muscle tissue for drug and tissue toxicity

循环系统和综合肌肉组织的药物和组织毒性

基本信息

批准号：
8668710
负责人：
George A Truskey
金额：
$ 15.7万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-07-24 至 2014-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8668710
关键词：
Beds Blood Vessels Caliber Carbon Dioxide Cardiac Output Cardiovascular system Cell Density Cells Computers Contracts Development Diabetes Mellitus Disease Drug toxicity Effectiveness Electrodes Electromagnetics Endothelial Cells Endothelium Evaluation Exercise Fiber Fibroblasts Gases Glucose Heart failure Human Hydrogels Inflammatory Laboratories Liquid substance Measurement Measures Mechanical Stimulation Mechanics Mediating Mesenchymal Stem Cells Metabolic Metabolism Methodology MicroRNAs Microcirculatory Bed Microfabrication Microfluidics Monitor Muscle Muscle Contraction Muscle Fibers Muscle function Muscular Dystrophies Myopathy Nutrient Organ Outcome Oxygen Oxygen Consumption Partial Pressure Performance Perfusion Peripheral arterial disease Pharmaceutical Preparations Phase Physiologic Monitoring Physiological Production Property Pulsatile Flow Pump Relative (related person)Research Resistance Role Running Severities Severity of illness Simulate Skeletal Muscle Smooth Muscle Myocytes Stimulus System Techniques Testing Thick Tissues Toxic effect Toxicity Tests Toxin U-Series Cooperative Agreements Vasoconstrictor Agents Vasodilator Agents Venous Venous system cerivastatin design drug candidate drug testing fatty acid metabolism glucose metabolism in vivo induced pluripotent stem cell metabolomics muscle engineering muscle form muscular system pressure research study response sarcopenia scaffold screening uptake vascular bed

项目摘要

DESCRIPTION (provided by applicant): Skeletal muscle is important for drug and toxicity testing given the relative size of the muscle mass and cardiac output that passes through muscle beds, the key role of muscle in energy substrate metabolism and diabetes, its role in mediating the severity of peripheral arterial disease and heart failure, and the need for therapies for muscle diseases such as muscular dystrophy and sarcopenia. To develop a system for functional and drug testing under physiological conditions, we will incorporate three-dimensional skeletal muscle cultures in a circulatory system that consists of a high-pressure arterial system that carries media to various tissue microcirculatory organ beds and returned via a low-pressure venous system. Arterial vessels will consist of an inner layer of endothelium and layers of differentiated vascular smooth muscle cells or mesenchymal stem cells. A computer controlled pump and valve system will pump small volumes of fluid to mimic arterial flow. Measurement of O2, CO2 and pressure will be used to control flow to the various beds. The modular design of the microcirculatory organ beds facilitates integration with a broad array of other organ and tissue mimics as part of the UH3 phase of the Cooperative Agreement. All experiments will use primary human cells. To generalize the applicability of the test bed, we will develop mature smooth muscle cells and skeletal muscle from iPS cells. In Aim 1, we will fabricate and test a branching network of small caliber blood vessels consisting of several layers of contractile human smooth muscle cells or mesenchymal stem cells and a confluent layer of endothelium. Flow rates, vessel distension and contraction, and the resistance of the microfluidic microcirculatory beds lined with endothelium will control the flow distribution to the different microcirculatory beds. In Aim 2, we will develop three-dimensional constructs of skeletal muscle and fibroblasts under tension. Different levels of oxygen partial pressure in the arterial and venous inflow lines will be used to produce a range of oxygen gradients. The muscle will be connected to posts containing a ferrogel that contracts under electromagnetic stimulation and thereby loads the muscle fibers. Endothelium will cover the outside of the three-dimensional muscle cultures, serving as an interface between the perfusion medium and skeletal muscle fibers. Oxygen gradients across the muscle layer will be controlled by cell density and thickness of the cell layer. We will fabricate an electrode system to electrically stimulate the fibers and measure force production. In Aim 3, we will combine the vascular and muscle units and run the unit for four weeks. Measurement of O2, CO2 and pressure will be used to control overall flow and regulate flow to the different beds. We will assess vessel dilation and muscle function. In Aim 4, the completed system will be used to test the effect of local release of vasodilators and vasoconstrictors on flow distribution, glucose metabolism and oxygen uptake by muscle. We will examine the response of blood vessels and muscle to an inflammatory stimulus. Metabolic profiling will be performed to simulate different physiological conditions and response to drugs and toxins.

描述（由申请人提供）：考虑到肌肉质量的相对大小和通过肌床的心输出量、肌肉在能量底物代谢和糖尿病中的关键作用、其在介导外周动脉疾病和心力衰竭的严重程度以及治疗的需要用于肌肉疾病，如肌营养不良症和肌少症。为了开发一个在生理条件下进行功能和药物测试的系统，我们将把三维骨骼肌培养物纳入一个循环系统中，该系统由高压动脉系统组成，该系统将介质输送到各种组织微循环器官床并通过低压动脉返回。压力静脉系统。动脉血管由内皮内层和分化的血管平滑肌细胞或间充质干细胞层组成。计算机控制的泵和阀门系统将泵送少量液体以模拟动脉血流。 O2、CO2 和压力的测量将用于控制流向各个床的流量。作为合作协议 UH3 阶段的一部分，微循环器官床的模块化设计有助于与广泛的其他器官和组织模拟物集成。所有实验都将使用原代人类细胞。为了推广测试台的适用性，我们将从 iPS 细胞中开发出成熟的平滑肌细胞和骨骼肌。在目标 1 中，我们将制造并测试小口径血管的分支网络，该网络由多层可收缩的人类平滑肌细胞或间充质干细胞和融合的内皮层组成。流速、血管扩张和收缩以及衬有内皮的微流体微循环床的阻力将控制流向不同微循环床的流量分布。在目标 2 中，我们将开发处于张力下的骨骼肌和成纤维细胞的三维结构。动脉和静脉流入管线中不同水平的氧分压将用于产生一系列氧梯度。肌肉将连接到含有铁凝胶的柱子上，铁凝胶在电磁刺激下收缩，从而对肌肉纤维施加负荷。内皮细胞将覆盖三维肌肉培养物的外部，充当灌注介质和骨骼肌纤维之间的界面。肌肉层上的氧气梯度将由细胞密度和细胞层厚度控制。我们将制造一个电极系统来电刺激纤维并测量力的产生。在目标 3 中，我们将结合血管和肌肉单元并运行该单元四个星期。 O2、CO2 和压力的测量将用于控制总体流量并调节流向不同床的流量。我们将评估血管扩张和肌肉功能。在目标 4 中，完整的系统将用于测试血管舒张剂和血管收缩剂的局部释放对流量分布、葡萄糖代谢和肌肉摄氧的影响。我们将检查血管和肌肉对炎症刺激的反应。将进行代谢分析以模拟不同的生理条件以及对药物和毒素的反应。