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Stabilization of desmoglein-2 binding rescues arrhythmia in arrhythmogenic cardiomyopathy
Camilla Schinner, … , Thomas D. Mueller, Jens Waschke
Camilla Schinner, … , Thomas D. Mueller, Jens Waschke
Published May 7, 2020
Citation Information: JCI Insight. 2020;5(9):e130141. https://doi.org/10.1172/jci.insight.130141.
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Research Article Cardiology

Stabilization of desmoglein-2 binding rescues arrhythmia in arrhythmogenic cardiomyopathy

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Abstract

Arrhythmogenic cardiomyopathy (AC) is a genetic disease causing arrhythmia and sudden cardiac death with only symptomatic therapy available at present. Mutations of desmosomal proteins, including desmoglein-2 (Dsg2) and plakoglobin (Pg), are the major cause of AC and have been shown to lead to impaired gap junction function. Recent data indicated the involvement of anti-Dsg2 autoantibodies in AC pathogenesis. We applied a peptide to stabilize Dsg2 binding similar to a translational approach to pemphigus, which is caused by anti-desmoglein autoantibodies. We provide evidence that stabilization of Dsg2 binding by a linking peptide (Dsg2-LP) is efficient to rescue arrhythmia in an AC mouse model immediately upon perfusion. Dsg2-LP, designed to cross-link Dsg2 molecules in proximity to the known binding pocket, stabilized Dsg2-mediated interactions on the surface of living cardiomyocytes as revealed by atomic force microscopy and induced Dsg2 oligomerization. Moreover, Dsg2-LP rescued disrupted cohesion induced by siRNA-mediated Pg or Dsg2 depletion or l-tryptophan, which was applied to impair overall cadherin binding. Dsg2-LP rescued connexin-43 mislocalization and conduction irregularities in response to impaired cardiomyocyte cohesion. These results demonstrate that stabilization of Dsg2 binding by Dsg2-LP can serve as a novel approach to treat arrhythmia in patients with AC.

Authors

Camilla Schinner, Bernd Markus Erber, Sunil Yeruva, Angela Schlipp, Vera Rötzer, Ellen Kempf, Sebastian Kant, Rudolf E. Leube, Thomas D. Mueller, Jens Waschke

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Figure 3

Disruption of intercellular junctions is rescued by stabilization of Dsg2 binding.

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Disruption of intercellular junctions is rescued by stabilization of Dsg...
(A) Dissociation assays of HL-1 monolayer treated with Dsg2- or Pg-siRNA and additional Dsg2-LP treatment (n = 8). One-way ANOVA with Bonferroni’s post hoc test. (B) Western blot analysis for Dsg2 and Pg performed in parallel to dissociation assays to show successful protein reduction after Dsg2- or Pg-siRNA treatment. α-Tubulin (α-Tub) served as loading control (n = 8). (C) Dissociation assay of HL-1 cell monolayer after disruption of cell cohesion by Trp or Dsg2-IP with Dsg2-LP treatment (control n = 8; Trp n = 8; Trp + Dsg-LP n = 7; Dsg2-LP n = 5; Dsg2-IP n = 3). One-way ANOVA with Bonferroni’s post hoc test. (D) Representative images of bead fragmentation after application of mechanical stress. Adhesion assay of Dsg2-Fc–coated beads with same conditions as C. (E) Ca2+ chelator EGTA and anti-Dsg2 antibody served as negative controls (control n = 10; EGTA n = 9; anti-Dsg2 n = 10; Trp n = 8; Trp + Dsg-LP n = 8; Dsg2-LP n = 8; Dsg2-IP n = 8). One-way ANOVA with Bonferroni’s post hoc test. (F) Representative immunostaining of HL-1 cells stained for Dsg2 (red) and Cx43 (green) treated with Trp, Dsg2-LP, or a combination of both for 24 hours (n = 8); scale bar: 10 μm. (G) Quantitative analysis of single-Dsg2 particle size (>30 cells from 3–4 independent experiments). (H) Quantitative analysis of Cx43 staining as ratio of intensity of staining at cell-cell border versus cytosol (>30 cells from 3 independent experiments). *P < 0.05. One-way ANOVA with Tukey’s post hoc test.

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