Cardiac-Cell Mechanics at the Single-Cell Level: Properties and Interactions

这份提案描述了一项为期五年的培训和研究计划，以扩大PI的电气 她拥有工程学背景，并帮助她获得了心脏细胞生物学方面的经验。为了实现这一点，PI (克莱姆森大学)将与她的导师，克莱姆森大学的Naren Vyavahare博士和Dr。 南卡罗来纳大学的托马斯·博格和伊迪·戈德史密斯博士。培训计划包括1) 细胞生物学和生物化学课程，2)心肌细胞实验技术培训 培养和分子生物学检测，以及3)通过讲习班和 人脉机会。这项培训的目的是让私人投资经理获得申请所需的技能 独立的NIH资金(例如，R01)，并使她成为一名平衡良好的生物医学工程师 研究员。 为了补充培训，PI提出了一项研究计划，旨在结合她的工程学 在她将接受培训的心脏细胞生物学技术方面的专业知识。细胞之间复杂的相互作用 功能和机械应力对许多生理和病理过程都很重要，例如 心脏等机械活动组织中的缺陷和疾病。对这一关系的深刻理解 将有助于更好地设计可用于组织工程化治疗的体外结构 机械活动的组织。目前，这些相互作用的性质还不完全清楚，这主要是由于 传统单细胞机械测量技术的局限性。拟议的研究重点是 了解作为微环境函数的心脏细胞力学和相互作用。首先， 心肌细胞和成纤维细胞的机械特性将作为细胞外基质的函数 使用原子力显微镜进行评估。然后，心肌细胞-成纤维细胞相互作用对细胞的影响 机制将会被确定。最后，将建立基于结构的细胞力学模型，这将使 使用来自成像技术的数据来预测心脏细胞的机械特性。 这项提议的长期目标是更好地了解心肌细胞的机械耦合。 这一理解对于建立有效的心脏组织功能的微观到宏观尺度模型至关重要， 这是开发基于再生医学的体内修复策略的关键。

This proposal describes a five-year training and research plan to broaden the Pi's electrical engineering background and help her gain experience in cardiac cell biology. To achieve this, the PI (Clemson University) will work closely with her mentors, Dr. Naren Vyavahare at Clemson University and Dr. Thomas Borg and Dr. Edie Goldsmith, at the University of South Carolina. The training plan includes 1) coursework in cell biology and biochemistry, 2) hands-on experimental technique training in cardiac cell culture and molecular biology assays, and 3) professional development activities through workshops and networking opportunities. The goal of this training to is allow the PI to gain the skills necessary to apply for independent NIH funding (e.g., R01) and to allow her to become a well-balanced biomedical engineering researcher. To complement the training, the PI proposes a research plan aimed at combining her engineering expertise with the cardiac cell biology techniques in which she will train. The complex interplay of cell function and mechanical stresses is important for many physiological and pathological processes, such as defects and diseases in mechanically active tissues like the heart. A solid understanding of this relationship would enable better design of in vitro constructs that could be used for tissue-engineered treatment of mechanically active tissues. Presently, the nature of these interactions is not fully understood, largely due to limitations in traditional single-cell mechanical measurement techniques. The proposed researchfocuses on understanding cardiac-cell mechanics and interactions as a function of the microenvironment. First, the mechanical properties of cardiomyocytes and fibroblasts as a function of the extracellular matrix will be assessed using atomic force microscopy. Then, the effect of cardiomyocyte-fibroblast interactions on cellular mechanics will be determined. Finally, structure-based cell-mechanics models will be built that will enable the use of data from imaging techniques to predict cardiac-cell mechanical properties. The long-term goal of this proposal is to better understand the mechanical coupling of cardiac cells. This understanding is critical for building effective micro- to macroscale models of cardiac tissue function, which is the key for developing in vivo repair strategies based on regenerative medicine.