Microtubules Increase Diastolic Stiffness in Failing Human Cardiomyocytes and Myocardium.

Background: Diastolic dysfunction is a prevalent and therapeutically intractable feature of heart failure (HF). Increasing ventricular compliance can improve diastolic performance, but the viscoelastic forces that resist diastolic filling and become elevated in human HF are poorly defined. Having recently identified post-translationally detyrosinated microtubules as a source of viscoelasticity in cardiomyocytes, we sought to test whether microtubules contribute meaningful viscoelastic resistance to diastolic stretch in human myocardium. Methods: Experiments were conducted in isolated human cardiomyocytes and trabeculae. First, slow and rapid (diastolic) stretch was applied to intact cardiomyocytes from non-failing and HF hearts, and viscoelasticity was characterized following interventions targeting microtubules. Next, intact left-ventricular trabeculae from HF patient hearts were incubated with colchicine or vehicle and subject to pre- and post-treatment mechanical testing, which consisted of a staircase protocol and rapid stretches from slack length to increasing strains. Results: Viscoelasticity was increased during diastolic stretch of HF cardiomyocytes compared to non-failing counterparts. Reducing either microtubule density or detyrosination reduced myocyte stiffness, particularly at diastolic strain rates, indicating reduced viscous forces. In myocardial tissue, we found microtubule depolymerization reduced myocardial viscoelasticity, with an effect that decreased with increasing strain. Colchicine reduced viscoelasticity at strains below, but not above, 15%, with a two-fold reduction in energy dissipation upon microtubule depolymerization. Post-hoc sub-group analysis revealed that myocardium from patients with HF with reduced ejection fraction (HFrEF) were more fibrotic and elastic than myocardium from patients with HF with preserved ejection fraction (HFpEF), which were relatively more viscous. Colchicine reduced viscoelasticity in both HFpEF and HFrEF myocardium. Conclusions: Failing cardiomyocytes exhibit elevated viscosity, and reducing microtubule density or detyrosination lowers viscoelastic resistance to diastolic stretch in human myocytes and myocardium. In failing myocardium, microtubules elevate stiffness over the typical working range of strains and strain rates, but exhibited diminishing effects with increasing length, consistent with an increasing contribution of the extracellular matrix and/or myofilament proteins at larger excursions. These studies indicate that a stabilized microtubule network provides a viscous impediment to diastolic stretch, particularly in HF.