EGFR inhibition leads to enhanced desmosome assembly and cardiomyocyte cohesion via ROCK activation
Maria Shoykhet, Orsela Dervishi, Philipp Menauer, Matthias Hiermaier, Sina Moztarzadeh, Colin Osterloh, Ralf J. Ludwig, Tatjana Williams, Brenda Gerull, Stefan Kääb, Sebastian Clauss, Dominik Schüttler, Jens Waschke, Sunil Yeruva
Maria Shoykhet, Orsela Dervishi, Philipp Menauer, Matthias Hiermaier, Sina Moztarzadeh, Colin Osterloh, Ralf J. Ludwig, Tatjana Williams, Brenda Gerull, Stefan Kääb, Sebastian Clauss, Dominik Schüttler, Jens Waschke, Sunil Yeruva
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Research Article Cardiology Cell biology

EGFR inhibition leads to enhanced desmosome assembly and cardiomyocyte cohesion via ROCK activation

Abstract

Arrhythmogenic cardiomyopathy (AC) is a familial heart disease partly caused by impaired desmosome turnover. Thus, stabilization of desmosome integrity may provide new treatment options. Desmosomes, apart from cellular cohesion, provide the structural framework of a signaling hub. Here, we investigated the role of the epidermal growth factor receptor (EGFR) in cardiomyocyte cohesion. We inhibited EGFR under physiological and pathophysiological conditions using the murine plakoglobin-KO AC model, in which EGFR was upregulated. EGFR inhibition enhanced cardiomyocyte cohesion. Immunoprecipitation showed an interaction of EGFR and desmoglein 2 (DSG2). Immunostaining and atomic force microscopy (AFM) revealed enhanced DSG2 localization and binding at cell borders upon EGFR inhibition. Enhanced area composita length and desmosome assembly were observed upon EGFR inhibition, confirmed by enhanced DSG2 and desmoplakin (DP) recruitment to cell borders. PamGene Kinase assay performed in HL-1 cardiomyocytes treated with erlotinib, an EGFR inhibitor, revealed upregulation of Rho-associated protein kinase (ROCK). Erlotinib-mediated desmosome assembly and cardiomyocyte cohesion were abolished upon ROCK inhibition. Thus, inhibiting EGFR and, thereby, stabilizing desmosome integrity via ROCK might provide treatment options for AC.

Authors

Maria Shoykhet, Orsela Dervishi, Philipp Menauer, Matthias Hiermaier, Sina Moztarzadeh, Colin Osterloh, Ralf J. Ludwig, Tatjana Williams, Brenda Gerull, Stefan Kääb, Sebastian Clauss, Dominik Schüttler, Jens Waschke, Sunil Yeruva

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Figure 2

EGFR inhibition leads to increased DSG2 binding frequency at cell borders.

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EGFR inhibition leads to increased DSG2 binding frequency at cell border...
(A–C) Binding frequency and topography during atomic force microscopy (AFM) measurements on HL-1 cardiomyocytes before and after erlotinib treatment at or close to cell borders and cell surfaces with tips coated with DSG2 (A), N-CAD (B), and DSC2 (C). Measurements were first performed in 90-minute DMSO-treated samples; then, medium was changed, erlotinib was added, and experiments were performed after 90 minutes. Green dots in topography images indicate binding events; cyan dots indicate cell borders. Topography images are 1.5 × 5 μm. In the graphs, per data point, 1,500 curves were analyzed across 2 areas (1.5 × 5 μm). *P ≤ 0.05. Statistical significance was calculated between DMSO versus erlotinib. Unpaired Student’s t test, n = 3 independent experiments. (D–F) Unbinding forces measured during AFM measurements on HL-1 cardiomyocytes at cell borders and cell surfaces with tips coated with DSG2 (D), N-CAD (E), and DSC2 (F). Per data point, 1,500 curves were analyzed across 2 areas (1.5 × 5 μm). *P ≤ 0.05. Statistical significance was calculated between DMSO versus erlotinib. Unpaired Student’s t test, n = 3 independent experiments.