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

描述(由申请人提供)： 这份提案描述了一项为期五年的培训和研究计划，以拓宽PI的电气工程背景，并帮助她获得心脏细胞生物学方面的经验。为了实现这一目标，PI(克莱姆森大学)将与她的导师密切合作，克莱姆森大学的Naren Vyavahare博士以及南卡罗来纳大学的Thomas Borg博士和Edie Goldsmith博士。培训计划包括1)细胞生物学和生物化学课程，2)心脏细胞培养和分子生物学分析方面的动手实验技术培训，3)通过研讨会和网络机会进行专业发展活动。这项培训的目标是让PI获得必要的技能，以申请独立的NIH资金(例如，R01)，并使她成为一名平衡的生物医学工程研究员。 为了补充培训，PI提出了一项研究计划，旨在将她的工程专业知识与她将接受培训的心脏细胞生物学技术结合起来。细胞功能和机械应力的复杂相互作用对于许多生理和病理过程是重要的，例如心脏等机械活动组织中的缺陷和疾病。对这一关系的扎实理解将有助于更好地设计可用于机械活性组织的组织工程治疗的体外结构。目前，由于传统的单细胞机械测量技术的局限性，这些相互作用的本质还没有被完全理解。这项拟议的研究侧重于了解作为微环境函数的心脏-细胞力学和相互作用。首先，将使用原子力显微镜评估心肌细胞和成纤维细胞作为细胞外基质的功能的机械特性。然后，将确定心肌细胞-成纤维细胞相互作用对细胞力学的影响。最后，将建立基于结构的细胞力学模型，该模型将能够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。这一认识对于建立有效的从微观到宏观的心脏组织功能模型至关重要，这是发展基于再生医学的体内修复策略的关键。

DESCRIPTION (provided by applicant): 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 research focuses 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.