Integrated System for Mechanoelectrical Studies of Cardiac Myofibroblasts

心脏肌成纤维细胞机电研究集成系统

• 批准号: 8311649
• 负责人: DANIEL H REICH
• 金额: $ 19.92万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-08-05 至 2014-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8311649
• 关键词: Actins; Adhesives; Advanced Development; Aleurites; Architecture; Arrhythmia; Biological Models; Calcium; Cardiac; Cell Shape; Cells; Cicatrix; Collaborations; Coupling; Cues; Development; Disease; Disease Progression; Duchenne muscular dystrophy; Electric Stimulation; Electrophysiology (science); Elements; Fibroblasts; Generations; Glaucoma; Glioma; Goals; Individual; Investigation; Joints; Laboratories; Link; Magnetism; Measurement; Measures; Mechanical Stimulation; Mechanics; Membrane; Membrane Potentials; Microfabrication; Monitor; Muscle; Myocardial Infarction; Myocardium; Myofibroblast; Nanotechnology; Pattern; Physics; Play; Polycystic Kidney Diseases; Printing; Process; Production; Proteins; Research; Research Personnel; Resolution; Role; Shapes; Signal Transduction; Source; Stress; Stress Fibers; System; Techniques; Technology; Time; Tissues; Traction; Universities; base; bone cell; cell growth; cell type; elastomeric; flexibility; innovation; insight; nanoparticle; nanowire; novel; response; sensor; tumorigenesis; voltage; voltage clamp

DESCRIPTION (provided by applicant): In diverse cell types such as muscle, fibroblasts and bone, the cells can generate contractile or traction forces while being responsive to electrophysiological and mechanical signals. Although subsets of these interactions have been studied under the themes of mechanotransduction and excitation-contraction coupling, a full understanding of the interrelationship between cell mechanics and cell electrophysiology requires the simultaneous measurement of cell force, cell voltage and cell current during the application of external forces. The goal of this project is to assemble, for the first time, an integrated measurement and force application system that will enable the functional role of mechanosensitive channels, transmembrane voltage, and specific membrane currents to be examined in cells having different cytoskeletal architectures. The model system that will be studied is the cardiac myofibroblast, a cell that plays a prominent role in myocardial infarcts and scar formation and is capable of force sensing, force production, and electrophysiological activity. In recent years, micropatterned flexible substrates, such as arrays of elastomeric micrometer-scale posts that deflect under cellular traction forces have emerged as novel systems for measuring cellular contractility with sub-cellular resolution. Further, by embedding magnetic nanoparticles in some of the posts, it is now possible to apply forces to adherent cells while measuring their mechanical responses via the surrounding non-magnetic posts. In this project, magnetic micropost-based force generation and measurement systems will be combined with electrophysiological techniques to enable simultaneous electrical and mechanical stimulation and readout of single cells. This project will be a joint effort between the laboratories of two Investigators with complementary expertise in cardiac electrophysiology, experimental physics, patterned cell growth, magnetism, microfabrication, and cell mechanics. Specific Aim 1 will integrate single cell traction force measurements via the micropost arrays with electrophysiological measurements of transmembrane potential and membrane ionic currents. Micro contact printing will be used to vary the shape of the myofibroblasts to manipulate their internal distribution of actin stress fibers, and to explore the effect of sub-cellular force distributions on transmembrane potential and currents. Specific Aim 2 will integrate magnetic actuation of mechanical forces and strains into the electromechanical readout system developed in Aim 1, and will study the mutual interactions of cellular mechanical and electrophysiological responses to external force and strain. By creating a unified technology for the study of cellular mechanoelectrical function, this project will open new directions for the elucidation of biologically and clinically important systems. )

描述（由申请人提供）：在肌肉，成纤维细胞和骨骼等各种细胞类型中，细胞可以产生收缩或牵引力，同时对电生理和机械信号响应。尽管这些相互作用的子集已在机械转移和激发诱导耦合的主题下进行了研究，但对细胞力学与细胞电生理学之间的相互关系的充分理解需要同时测量外部力期间细胞力，细胞电压和细胞电流。该项目的目的是首次组装一个集成的测量和力应用系统，该系统将使机械敏感通道，跨膜电压和特定的膜电流的功能作用在具有不同细胞骨架骨骼架构的细胞中进行检查。将研究的模型系统是心脏肌纤维细胞，该细胞在心肌梗死和疤痕形成中起着重要作用，并且能够感应力，力产生和电生理活性。近年来，微图案的柔性底物，例如在细胞牵引力下偏转的弹性微米尺度柱阵列已成为用于用亚细胞分辨率测量细胞收缩力的新型系统。此外，通过将磁性纳米颗粒嵌入某些柱子中，现在可以将力施加到粘附细胞上，同时通过周围的非磁性柱测量其机械响应。在这个项目中，将基于磁性微型的力产生和测量系统与电生理技术结合使用，以同时对单细胞进行电气和机械刺激和读数。该项目将是两个研究人员的实验室共同努力，这些研究人员在心脏电生理学，实验物理学，图案化细胞生长，磁性，微结构和细胞力学方面具有互补的专业知识。特定的目标1将通过微型矩阵将单细胞牵引力测量与跨膜电位和膜离子电流的电生理测量结果相结合。微接触打印将用于改变肌纤维细胞的形状，以操纵其内部肌动蛋白应力纤维的分布，并探索亚细胞力分布对跨膜电位和电流的影响。特定的目标2将将机械力和应变的磁性致动纳入AIM 1中开发的机电读出系统，并将研究细胞机械和电生理对外力和应变的相互相互作用。通过创建统一的技术来研究细胞机械功能，该项目将为阐明生物学和临床重要的系统打开新的方向。 ）