Cellular mechanical properties, collectively referred to as mechanotype, play a role in cell physiology and pathology, including cell proliferation, survival, metabolism, stem cell differentiation, immune cell migration, and cancer metastasis. Cell deformability and contractility are two key characteristics that determine the mechanotype of a cell. We have focused on understanding how cellular mechanotype is regulated by microenvironmental inputs that have been implicated in cell invasion, such as glucose levels. Hyperglycemia (HG) is prevalent in obesity and diabetes, which in turn are factors facilitating cancer progression. The effects of HG on cellular mechanotype are the focus of this project. The mentored principal investigator (mPI) has developed a novel cell mechanotyping tool to probe cell deformability, called parallel microfiltration (PMF). In this project, using PMF and related technologies, we will define effects of glucose on cell mechanotype in two distinct model systems: breast cancer cells and macrophages. Our hypothesis is that the HG effects on cellular mechanotype have critical consequences on cell migration, invasiveness, and anoikis. Our long-term objective is to identify pathways that regulate cell mechanotype, migration, and survival under HG conditions, which is of translational relevance and health significance in the context of cancer and immune responses. The specific aims are: 1. Determine how glucose regulates the mechanotype of cancer and immune cells. In this aim, we will define the mechanistic basis of glucose-mediated mechanotype regulation that results in alterations in cell migration. We will use two models: (i) breast cancer cells; and (ii) macrophages. We will employ a novel mechanotyping technique invented by the mPI and collaborators. 2. Delineate how glucose-mediated mechanotype alterations affect cell survival. In this aim, we will determine how regulators and mediators of mechanotype dynamics influence anoikis of cancer cells.