Despite decades of advances in clinical diagnosis and treatment of heart disease, there is a rising epidemic of heart failure in an aging population worldwide. Historically, much focus has been on atherosclerosis, acute myocardial infarction, and treatment of large vessel coronary disease. However, relatively little is known at a mechanistic level about the vast network of small arteries, capillaries and veins in the heart beyond the large coronary arteries, the coronary microcirculation, and increasing evidence links dysfunction of the microcirculation to various forms of heart disease including heart failure. In vitro experiments, histologic analysis, and computational modeling have provided insight about how the coronary microcirculation is regulated and remodels in the failing heart, but prior studies have been unable to directly evaluate the complex microcirculatory physiology of the heart at the cellular level in vivo. Intravital optical microscopy is being used in the neurosciences and tumor biology to decipher dynamic vascular physiology in vivo, but these techniques have not been applicable in the heart due to severe imaging limitations imposed by contractile motion. We have recently pioneered intravital imaging methods to perform motion-artifact free, cellular resolution microscopy in the beating heart. Building on this work, this proposal seeks to investigate mechanisms of coronary microvascular dysfunction by utilizing intravital microscopy to quantitatively map ?ow in the coronary microcirculation down to the capillary level in animal models of heart disease. Studies will be performed in mice comparing microcirculatory function in healthy controls, in a model of physiologic hypertrophy due to exercise, and in a model of pressure overload leading to pathologic hypertrophy and heart failure. In addition, the speci?c role of microvascular pericytes will be investigated as a master regulator of capillary blood ?ow. Intravital confocal and two-photon microscopy, multiplexed ?uorescent reporters, and cell-speci?c optogenetics approaches will be used to record and manipulate microvascular ?ow, and to correlate abnormal ?ow patterns with cardiomyocyte metabolism, in?ammatory response, and ?brosis. This work will lead to new understanding of microcirculatory pathophysiology in the failing heart at the single cell level and promising new insights for clinical therapeutics.
This project investigates fundamental mechanisms of coronary microvascular dysfunction at the cellular level using high resolution intravital microscopy in animal models of heart disease. The work promises to provide new insights into mechanisms of heart failure and will inform new approaches to diagnosis and treatment. It will also lead to innovations in high resolution imaging of the heart that will empower future studies in the cardiovascular sciences.