CAREER: Investigating the Cellular Electrome as a Biomarker in Red Blood Cell Physiology and Pathology

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Red blood cells (RBCs) are the most abundant cell in the human body and are responsible for carrying oxygen from our lungs to the rest of our body, thus it is crucial to understand how these cells work especially in diseases that affect their well-being. This work seeks to address critical gaps in knowledge of RBCs using a variety of techniques that will probe internal, external, and membrane alterations due to changes in RBC health. This will elicit further understanding of early changes in RBC-related diseases such as malaria, anemia, and cardiovascular diseases. In addition to progressing research, this CAREER award will educate the future workforce as well as the community, who need to make informed decisions regarding new technologies. This project will teach students research concepts, expose them to novel cell characterization and state-of-the-art technologies, and train them how to communicate their work with the public. Current red blood cell (RBC) analysis often relies on manual observations to changes in cell morphology or behavior. Electrical measures of a cell contain vital information about their function(s). They are, however, difficult to measure. Standard techniques typically measure no more than a few cells and require skilled technicians to complete the analysis. Further, no technique accounts for all ionic currents present in a cell. Still, broader impact of these measurements remains high, particularly in drug discovery and disease progression. The objective of this CAREER proposal is to investigate the existence of an interconnected “cell electrome” in the analysis of RBCs and RBC health monitoring. As the proteome is the complete set of proteins expressed by a cell, the “electrome” represents the complete set of ionic currents present in a cell and includes cell surface zeta-potential, membrane potential, and Dielectrophoresis (DEP)-measured membrane conductance, membrane capacitance, and cytoplasm conductivity. Dielectrophoresis (DEP) is an electrokinetic technique that relies on the electrophysiological properties of cells and the suspending media as well as the geometry of the applied electric field to induce movement of these cells. Since DEP does not rely on biomarkers or other fluorescent labels, it has become an increasingly popular method of cell characterization. The primary hypothesis of this work is that DEP parameters can robustly describe changes in RBC electrome due to oxidative stress, a dominant factor in RBC-related diseases, providing a rapid and label-free biomarker. How these parameters are linked will increase understanding of RBC electrophysiology and its role in RBC-related diseases. The electrome, once understood in RBCs, could be expanded to other, more complicated cell types and may also explain behaviors in these other cells such as electrophysiology alterations to influence cell-to-cell interactions in metastasizing cancer.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.