A Nanomaterial-Engineered Proteomic Platform to Map Curvature-Dependent Cardiac Signaling

About This Grant

PROJECT SUMMARY Membrane curvature clusters the signaling receptors and ion channels that govern cardiac excitation–contraction coupling. In cardiomyocytes, sharply curved caveolae (~60 nm), moderately curved clathrin-coated pits (~120 nm), and cylindrically curved transverse tubules (T-tubules) modulate β-adrenergic and Ca2+ pathways through their distinct protein sets. Interruptions to their shape and protein composition lead to arrhythmia, hypertrophy, or contractile failure, respectively. Current imaging and proteomic tools capture only fragments of these nanodomains, leaving a technological gap in unbiased curvature-defined proteomics and a knowledge gap in how geometry dictates cardiac signaling. The overall objective of this project is to close these gaps by developing a Nanomaterial-Engineered Proteomic (NanoPro) platform and providing a proteomic atlas for each curvature class in cardiomyocytes. NanoPro uses nanostructures to proximity-label nearby proteins on the cell membrane and produces unbiased, 20 nm-resolution proteomic atlases of each curvature class. We will reach the objective through two specific aims. Aim 1 will establish dome-NanoPro to profile the proteome of nanomaterial-engineered caveolae and clathrin-coated pits in cardiomyocytes. Aim 2 will streamline pillar-NanoPro to map the proteome of nanomaterial-engineered T-tubules in cardiomyocytes. NanoPro represents a new concept of structure-defined proteomics by integrating cell membrane engineering with deep, unbiased proximity-labeling mass spectrometry. Upon successful development, NanoPro will close the technological gap in curvature-defined proteomics and narrow the long-standing knowledge gap in cardiac cell signaling by revealing how membrane curvature orchestrates signaling networks in caveolae, clathrin-coated pits, and T-tubules with the first complete proteomic atlas of these structures. These proteomic data sets will expose curvature-encoded biomarkers for early detection of cardiac stress and uncover druggable targets that can be regulated to prevent arrhythmia, hypertrophy, and contractile failure.