DMS/NIGMS1: Unravel the connection between deformability and motility of eukaryotic cell through a - Cell migration, an inherently complex process involving a multitude of biological mechanisms, plays a pivotal role in physiological, developmental, and disease-related processes, such as cancer metastasis, wound healing, and immune responses. Despite its importance, our understanding of the relationship between cell stiffness and migration remains incomplete. While diverse tools and methods available for probing cell mechanics and migration have enabled researchers to approach this subject from multiple perspectives, each method operates under a specific set of parameters, rules, and rheological metrics, making direct comparisons between studies nearly impossible. Another limitation of existing approaches is that they fail to provide information about cell stiffness in situ and thus cannot directly correlate with cell migration. To address the gaps in our knowledge and methodologies, we propose to build a combined in vitro and in silico multi-modal platform to unravel the connection between the stiffness and motility of eukaryotic cells. The in vitro platform contains custom-designed high-throughput microfluidic chips that sort cells based on their stiffness. Cells sorted into different stiffness groups can be retrieved for subsequent cell motility tests driven by chemotaxis. In addition, immunofluorescence staining, single-cell western blotting, and atomic force microscopy measurement will be performed on the single-cell level to probe the correlation among the levels of various structural and adhesion proteins, cell stiffness, and motility. The in silico platform consists of computational models serving as digital counterparts of the cells and microfluidic devices to efficiently connect single-cell analysis and population study. After validation using data generated from the in vitro platform, these computational models will be used to dissect the contribution of internal components of eukaryotic cells to their stiffness and ability to migrate, thereby providing mechanistic rationale for experimental findings and offering new insights into the understanding of various disease-related processes. The impact of the proposed in vitro and in silicon multi-modal platform is significant because it is expected to provide a standardized platform to analyze and sort cells based on their stiffness, investigate single-cell migration for cells with specific stiffness, and evaluate the motility and stiffness-related proteins at the single- cell level for a variety of cell types. RELEVANCE (See instructions): In this application, we will build and validate a combined in vitro and in silico platform that enables us to investigate the stiffness and motility of a variety of cell types, such as tumor cells, phagocytes and fibroblasts, thereby advancing our knowledge of how the stiffness of cells impacts their ability to migrate within the tissue.
