Myosin in autoinhibitedoffstate(s), stabilized by mavacamten, can be recruited via inotropic effectors

Abstract

AbstractMavacamten is a novel, FDA-approved, small molecule therapeutic designed to regulate cardiac function by selectively but reversibly inhibiting the enzymatic activity of myosin. It shifts myosin towards orderedoffstates close to the thick filament backbone. It remains unresolved whether mavacamten permanently sequesters these myosin heads in theoffstate(s) or whether these heads can be recruited in response to physiological stimuli when required to boost cardiac output. We show that cardiac myosins stabilized in theseoffstate(s) by mavacamten are recruitable by Ca2+, increased heart rate, stretch, and β-adrenergic (β-AR) stimulation, all known physiological inotropic effectors. At the molecular level, we show that, in presence of mavacamten, Ca2+increases myosin ATPase activity by shifting myosin heads from the reserve super-relaxed (SRX) state to the active disordered relaxed (DRX) state. At the myofilament level, both Ca2+and passive lengthening can shift orderedoffmyosin heads from positions close to the thick filament backbone to disorderedonstates closer to the thin filaments in the presence of mavacamten. In isolated rat cardiomyocytes, increased stimulation rates enhanced shortening fraction in mavacamten-treated cells. This observation was confirmedin vivoin telemetered rats, where left-ventricular dP/dtmax,an index of inotropy, increased with heart rate in mavacamten treated animals. Finally, we show that β-AR stimulationin vivoincreases left-ventricular function and stroke volume in the setting of mavacamten. Our data demonstrate that the mavacamten-promotedoffstates of myosin in the thick filament are activable, at least partially, thus leading to preservation of cardiac reserve mechanisms.Significance statementMavacamten is the first myosin-targeted small molecule inhibitor approved by the FDA to treat obstructive hypertrophic cardiomyopathy by attenuating myocardial hyperdynamic contraction. The recruitment of cardiac contractility is, however, vital to ensure sufficient cardiac output during increased physiological demand. Here we show that major inotropic effectors are at least partially preserved in the setting of mavacamten, resulting in maintenance of cardiac reserve mechanisms. These results not only suggest an alternative mechanistic explanation, beyond mere LV outflow tract obstruction removal, for the clinically observed increase in peak oxygen uptake with exercise in HCM patients receiving mavacamten, but also lay the groundwork for a potential methodology to investigate the sarcomeric basis of chronotropic incompetence in disease states to motivate new therapeutic interventions.