Cell type specific CaMKII activation patterns revealed by CaMKAR, a bioactivity reporter deployable in living cells

Abstract

An accurate and precise mechanism for measuring CaMKII activity in living cells is invaluable in the search for effective and targeted CaMKII-based therapeutics. Here, we employ our recently published CaMKII Activity Reporter (CaMKAR) biosensor in order to investigate the spatiotemporal dynamics of CaMKII activation in three different types of cells - cardiac myocytes, skeletal myocytes, and neurons. In doing so, we found a greater rate of CaMKII activation in skeletal muscle compared to cardiac muscle and also delineated CaMKARs ability to measure discrete CaMKII activation events in the presence of individual action potentials. By modifying the original CaMKAR sequence, we generated sensors that can be localized to subcellular compartments and thereby preferentially detect the activity of specific spatially-distributed CaMKII isoforms. Finally, we utilized the live-cell data to generate mathematical models of CaMKII activation kinetics, both as an integrated function across multiple calcium transients and as discrete on-off events following individual depolarizations. By furthering our understanding of CaMKII activity profiles across cell types and within subcellular compartments, we hope to support development of CaMKII inhibitors that are optimally precise and potent.