Multiscale spatiotemporal modeling of cardiac mitochondria

描述(申请人提供)：线粒体是一种丰富而重要的亚细胞细胞器，其组织和超微结构复杂，但无法解释。这一多PI多尺度建模方案试图使用数学建模和生物学实验相结合的方法来检验心肌线粒体纳米超微结构是线粒体功能最终成功的原因这一总体假设。大鼠心肌细胞线粒体将用于计划中的建模和生物学实验。我们的初步实验和公布的数据表明，计划的工作是可行的，三个PI可以成功地完成这一雄心勃勃和具有挑战性的提案。新鲜分离的大鼠心肌细胞和短期(1-3天)培养的大鼠心肌细胞将在Leder实验室制备，由Mannella团队进行电子显微镜(EM)断层扫描，并通过三种PI进行定量和交互分析。莱德尔团队将使用共聚焦和超分辨率随机光学重建显微镜(STORM)对活细胞进行检查。从EM断层扫描和活细胞成像获得的定量空间和功能数据将用于通知和约束以Jafri实验室为中心的线粒体的多尺度3D建模。计划中的生物实验和数学建模的迭代方法将使这项研究首次确定线粒体功能的结构基础。最后，这项拟议的研究试图包括在控制条件下对线粒体进行建模，当线粒体受到简单干预的压力时，以及当线粒体因压力超负荷心力衰竭而改变时。因此，这项工作不仅将为线粒体如何发挥功能提供基本的新信息，还将为线粒体相关疾病的新疗法奠定基础。

DESCRIPTION (provided by applicant): Mitochondria are abundant and critically important subcellular organelles whose organization and ultrastructure are complex but unexplained. This multi-PI multiscale modeling proposal seeks to use a combination of mathematical modeling and biological experiments to test the overall hypothesis that the nanoscopic ultrastructure of cardiac mitochondria accounts for the ultimate success of mitochondrial function. Mitochondria in rat cardiac ventricular myocytes will be used in the planned modeling and biological experiments. Our preliminary experiments and published data indicate that the planned work is feasible and that the three PIs can succeed in this ambitious and challenging proposal. Freshly isolated rat cardiac ventricular myocytes and those in short-term (1-3 days) culture will be prepared in the Lederer lab and imaged with electron microscope (EM) tomography by the Mannella team and analyzed quantitatively and interactively by the three PIs. Living cells will be examined by the Lederer team using confocal and super-resolution Stochastic Optical Reconstruction Microscopy (STORM). The quantitative spatial and functional data obtained from EM tomography and live cell imaging will be used to inform and constrain the multi-scale 3D modeling of mitochondria centered in the Jafri lab. The planned iterative approach to biological experiments and mathematical modeling will enable this investigation to define the structural basis for mitochondrial function for the first time. Finally, the proposed investigation seeks to include modeling of mitochondria under control conditions, when mitochondria are stressed by simple interventions and when mitochondria are altered by pressure-overload heart failure. This work therefore will not only provide fundamental new information on how mitochondria function but will also lay the foundation for novel therapies in mitochondrially involved diseases.