In vitro models incorporating the complexity and function of adult human tissues are highly desired for translational research. Whilst vital slices of human myocardium approach these demands, their rapid degeneration in tissue culture precludes long-term experimentation. Here, we report preservation of structure and performance of human myocardium under conditions of physiological preload, compliance, and continuous excitation. In biomimetic culture, tissue slices prepared from explanted failing human hearts attain a stable state of contractility that can be monitored for up to 4 months or 2000000 beats in vitro. Cultured myocardium undergoes particular alterations in biomechanics, structure, and mRNA expression. The suitability of the model for drug safety evaluation is exemplified by repeated assessment of refractory period that permits sensitive analysis of repolarization impairment induced by the multimodal hERG-inhibitor pentamidine. Biomimetic tissue culture will provide new opportunities to study drug targets, gene functions, and cellular plasticity in adult human myocardium.

Taken together, we provide first evidence for positive adhesiotropy as a new cardiac function of sympathetic signaling. Positive adhesiotropy is dependent on Pg phosphorylation at S665 by protein kinase A. This mechanism may be of high medical relevance because loss of junctional Pg is a hallmark of arrhythmogenic cardiomyopathy.

Our data demonstrate that desmoglein-2 plays a critical role in cardiomyocyte cohesion and function.

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Desmosomal proteins including desmoglein‐2 (DSG2), plakoglobin (PG), and desmoplakin (DP) are components of the intercalated disc and mediate cardiac myocyte cell‐cell adhesion. Enhancement of cardiac myocyte cohesion, referred to as “positive adhesiotropy”, was demonstrated to be a function of sympathetic signaling in the heart and to be relevant for a sufficient positive inotropic response to adrenergic stimulation. Here we used the positive inotropic agent digitoxin to further investigate the link between inotropy and adhesiotropy in cardiac myocytes. Digitoxin failed to enhance cardiac contractility in Langendorff‐perfused mouse hearts lacking the desmosomal plaque protein PG indicating that a positive inotropic response requires intact desmosomal adhesion. To investigate a potential effect of digitoxin on desmosomal adhesion, atomic force microscopy was applied and revealed that digitoxin increased the binding force of the adhesion molecule DSG2 at cell‐cell contact areas. This was paralleled by enhanced cardiac myocyte cohesion in both HL‐1 cardiac myocytes and murine cardiac slices as determined by dissociation assays as well as by accumulation of DSG2, DP and PG at cell‐cell contact areas. However, total protein levels or cytoskeletal anchorage of desmosomal proteins was not affected. siRNA‐mediated depletion of DSG2, PG or DP abrogated increase of cell cohesion demonstrating that intact desmosomal adhesion is required for positive adhesiotropy. Mechanistically, digitoxin caused activation of ERK1/2 but not of p38MAPK or Src. In line with this, inhibition of ERK1/2 signaling using the inhibitor UO 126 abrogated the effects of digitoxin on cell‐cell adhesion and desmosomal reorganization. These results show that the positive inotropic agent digitoxin enhances cardiac myocyte cohesion with reorganization of desmosomal proteins in an ERK1/2‐dependent manner. Desmosomal adhesion seems to be important for a sufficient positive inotropic response of digitoxin treatment which can be medical relevant for the treatment of heart failure. Support or Funding Information This work was supported by the Ludwig‐Maximilians‐Universität Munich with the Wifomed program and the Deutsche Forschungsgemeinschaft DFG [grant number WA2474/11‐1 to J.W.] This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.