Mapping the Mechanome: A Systems-level Understanding of Mechanical Mediators and Pathways

About This Grant

ABSTRACT The central goal of this application is to harness cellular mechanobiology to achieve predictive control over cellular behaviors that are critical for physiological and disease contexts. A growing number of diseases are attributed to dysfunction in cellular mechanical regulation; mechanical and physical cues are now known to regulate cellular behaviors in diverse contexts from muscle regeneration to cancer progression to cellular reprogramming. Although specific mechanical signaling mediators and pathways have been well-studied at molecular and cellular scales, these mechanistic studies use a bottom-up approach that focuses on a single protein or pathway in a single cell type. However, cells integrate signals from various sources across timescales and length-scales to produce a coordinated response, which requires a top-down or systems-level understanding of the ‘mechanome’—the set of genes, proteins, and pathways that regulate cellular mechanotype—and represents a major gap in the field. If we had a systems-level understanding of the mechanome this would enable fundamental knowledge of cellular mechanobiology. Such knowledge of the mechanome would allow us to intervene and control the time-dependent trajectories of cells and is crucial for effective interventions for human health as well as engineering self-assembling cellular structures and tissues with spatio-temporal control. Building on my lab’s recent research achievements and new preliminary data, the Rowat lab will pursue two future lines of research: 1) To identify novel mediators of cell mechanical behaviors, we will take an unbiased approach using the high throughput cell filtration platform that my lab has developed to conduct a genome-wide pooled shRNA screen. Follow up studies will validate the top hits using complementary measurements of cell mechanics, force generation, and motility across cell types. We will further investigate mechanisms of how top candidates as well as the novel mechanical mediator, NUDT5, which we previously identified in an initial deformability screen, mediate the mechanical behaviors of fibroblasts and immune cells. 2) To define mediators of mechanical memory, we will engineer a small molecule screen to identify molecules that mediate cellular mechanical behaviors that persist over generations after mechanical priming, using cellular morphology as a readout for cells plated on a compliant substrate following culture on a stiff substrate. Follow up studies will define mechanisms of how the top candidate mediators control mechanical memory by quantifying effects on cell mechanical behaviors and subcellular phenotypes. Taken together, findings will provide a platform for generating unbiased, systems-level knowledge of the mechanome. Findings will also provide the foundation for an interactive, web-based mechanome atlas, a resource that will be accessible to broader research communities.