Buccal Mucosa Cells as a Potential Diagnostic Tool to Study Onset and Progression of Arrhythmogenic Cardiomyopathy

Figures

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  Figure 1. Examples of buccal mucosa scoring. A score of 4 means extreme strong membrane labelling and a score of 0 means no labelling at all. Scoring was performed on clusters consisting of more than three cells. Right bottom panel depicts a H&E staining of buccal mucosa cells. H&E; hematoxylin and eosin staining.

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  Figure 2. (A) Panel shows that there is no difference between membrane labelling score between males (n = 14) and females (n = 20) in any of the dilutions. (B) Depicts correlations between age and plakoglobin score. None of the dilutions show significant or biological relevant correlations with age. 1:5000 n = 31, 1:10,000 n = 32, 1:20,000 n = 32, 1:40,000 n = 33. rs; Spearman’s rho, n.s.: not significant. • indicate individual(s) for the correlation.

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  Figure 3. Examples of buccal mucosa smears of healthy controls and ACM patients labeled for plakoglobin at different dilutions. ACM; arrhythmogenic cardiomyopathy.

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  Figure 4. A heat map visualizing the plakoglobin score of all individual subjects (healthy controls n = 34, borderline ACM n = 14 and ACM diagnosed patients n = 19) for all four dilutions. A clear pattern is visible where healthy controls in general tend to show higher scores than borderline and ACM-diagnosed patients. Colors correspond to the score as shown in the right panel. At the left axis, controls (C) and ACM patient (A) numbers are indicated. Furthermore, for each ACM patient their TFC score is listed. ACM; arrhythmogenic cardiomyopathy.

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  Figure 5. (A) Strong and significant differences at the population level between healthy controls (n = 34), preclinical carriers (n = 14) and ACM diagnosed patients (n = 19) in all four dilutions. (B) Spearman correlation graphs for plakoglobin score and TFC are shown. (C) There is a strong significant correlation with premature ventricular contractions in ACM patients (n = 11). (D) Comparison of the sensitivity and specificity score of each dilution used in this study. TFC; Task Force Criteria score 2010, rs; Spearman’s rho. • indicate individual(s) for the correlation or represent dilution factor in figure D. **** p < 0.0001, *** p < 0.001, ** p < 0.01.

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  Figure 6. A heat map visualizing plakoglobin scores of all individual healthy controls (n = 34) and PLN carriers (n = 17), for all four dilutions. No clear difference was found between controls and PLN carriers in plakoglobin score. Colors correspond to score as shown in the right panel. At the left axis, controls (C) and PLN p.Arg14del patient (P) numbers are indicated. Furthermore, for PLN patients their clinical diagnosis is indicated when specified. PLN; phospholamban, ACM; arrhythmogenic cardiomyopathy, DCM; dilated cardiomyopathy.

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  Figure 7. (A) No significant differences at the population level between healthy controls (n = 34) and PLN carriers (n = 17) in any of the dilutions. (B) Graphs depict differences in plakoglobin score between controls (n = 34), PLN variant carriers that are preclinical (n = 7), with an ACM (n = 3) or DCM (n = 7) phenotype. ACM PLN patients seem to adhere more to the ACM patients’ pattern of plakoglobin score. PLN; phospholamban, ACM; arrhythmogenic cardiomyopathy, DCM; dilated cardiomyopathy. • indicate individual(s) for the correlation.