EGFR inhibition leads to enhanced desmosome assembly and cardiomyocyte cohesion via ROCK activation
Maria Shoykhet, … , Jens Waschke, Sunil Yeruva
Maria Shoykhet, … , Jens Waschke, Sunil Yeruva
Published February 16, 2023
Citation Information: JCI Insight. 2023;8(6):e163763. https://doi.org/10.1172/jci.insight.163763.
View: Text | PDF
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

×

Figure 8

Erlotinib-enhanced cardiomyocyte cohesion and desmosomal assembly are mediated by ROCK in HL-1 cardiomyocytes.

Options: View larger image (or click on image) Download as PowerPoint
Erlotinib-enhanced cardiomyocyte cohesion and desmosomal assembly are me...
(A) RhoA G-LISA showing enhanced RhoA activity upon EGFR inhibition by erlotinib. *P ≤ 0.05, unpaired Student’s t test, n = 3. (B) Representative Western blot showing phosphorylation of the ROCK target MLC2 upon erlotinib treatment. *P ≤ 0.05, 2-way ANOVA with Holm Sidak’s multiple comparison test, n = 6 independent experiments. (C) Dispase-based dissociation assay in HL-1 cardiomyocytes, after treatment with erlotinib with and without Y27632. Y27632 was added 30 minutes prior to erlotinib incubation for 60 minutes, with representative pictures of the wells. *P ≤ 0.05, 2-way ANOVA with Holm Sidak’s multiple comparison test, n = 8 independent experiments. (D) Immunostaining of DP and DSG2 in HL-1 cardiomyocytes with WGA as membrane marker after erlotinib treatment with and without Y26732, as in C. White arrows indicate areas of increased DP or DSG2 recruitment to the cell membrane. Scale bar: 10 μm. (E) Dispase-based dissociation assay in HL-1 cardiomyocytes after 90 minutes of Ca2+ depletion and treatment with erlotinib with and without Y26732 with representative pictures of the wells. *P ≤ 0.05, 2-way ANOVA with Holm-Sidak’s multiple comparison test, n = 7 independent experiments.