Cardiac safety assessment of a novel recombinant bispecific antibody targeting the ether-à-go-go related gene 1 (hERG1)-β1 integrin macromolecular complex

Figures

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  FIGURE 1. Action potential kinetics characterization of atrial cardiomyocytes isolated from valve-disease patients and incubated with hERG1-mAB, scFv-hERG1 and scDB-hERG1-β1. (A) Action potential duration at 90% repolarization (APD90%) recorded during stimulation at 0.2 Hz (left), 0.5 Hz (center) and 1 Hz (right) in human atrial cardiomyocytes incubated with hERG1-mAB (10 μg/mL), scDB-hERG1-β1 (10 μg/mL) or incubated with vehicle. (B) Superimposed representative action potential traces recorded in human atrial cardiomyocytes recorded during stimulation at 0.2 Hz (left), 0.5 Hz (center) and 1 Hz (right) and incubated with hERG1-mAB (10 μg/mL), scDB-hERG1-β1 (10 μg/mL) or vehicle. Mean ± SE from 24 cardiomyocytes (10 patients). * = p < 0.05; NS: not significant. (C) Action potential duration at 90% repolarization (APD90%) recorded during stimulation at 0.2 Hz (left), 0.5 Hz (center) and 1 Hz (right) in human atrial cardiomyocytes incubated with scFv-hERG1 (10 μg/mL), scDB-hERG1-β1 (10 μg/mL) or vehicle. (D) Superimposed representative action potential traces recorded in human atrial cardiomyocytes recorded during stimulation at 0.2 Hz (left), 0.5 Hz (center) and 1 Hz (right) and incubated…

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  FIGURE 2. Action potential kinetics characterization of ventricular cardiomyocytes isolated from patients with aortic stenosis (AOS) and incubated with hERG1-mAB, scFv-hERG1 and scDB-hERG1-β1. (A) Action potential duration at 20%, 50% and 90% repolarization (APD20%, APD50% and APD90%) recorded during stimulation at 0.2 Hz in human ventricular cardiomyocytes incubated with hERG1-mAB (10 μg/mL), scDB-hERG1-β1 (10 μg/mL) or not incubated with Abs. (B) Superimposed representative action potential traces recorded in human ventricular cardiomyocytes during stimulation at 0.2 Hz and incubated with hERG1-mAB (10 μg/mL), scDB-hERG1-β1 (10 μg/mL) or vehicle. (C) Action potential duration at 20%, 50% and 90% repolarization (APD20%, APD50% and APD90%) recorded during stimulation at 0.5 Hz in human ventricular cardiomyocytes incubated with hERG1-mAB (10 μg/mL), scDB-hERG1-β1 (10 μg/mL) or vehicle. (D) Superimposed representative action potential traces recorded in human ventricular cardiomyocytes during stimulation at 0.5 Hz and incubated with hERG1-mAB (10 μg/mL), scDB-hERG1-β1 (10 μg/mL) or with vehicle. Mean ± SE from 24 cardiomyocytes (8 patients). * = p < 0.05; NS: not significant. (E) Action…

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  FIGURE 3. Effect of E4031 on action potential kinetics evaluated in ventricular cardiomyocytes isolated from patients with aortic stenosis (AOS) and incubated with scDB-hERG1-β1 and scFv-hERG1. (A) Action potential duration at 20%, 50% and 90% repolarization (APD20%, APD50%, APD90%) recorded during stimulation at 0.2 Hz (left), 0.5 Hz (center) and 1 Hz (right)in human ventricular cardiomyocytes incubated with scDB-hERG1-β1 (10 μg/mL), in absence and presence of E4031 (5 µM). (B) Superimposed representative action potential traces recorded in human ventricular cardiomyocytes during stimulation at 0.2 Hz (left), 0.5 Hz (center) and 1 Hz (right) and incubated with scDB-hERG1-β1 (10 μg/mL), in absence and presence of E4031 (5 µM). Mean ± SE from 10 cardiomyocytes (4 patients). ** = p < 0.01; NS: not significant. (C) Action potential duration at 20%, 50% and 90% repolarization (APD20%, APD50%, APD90%) recorded during stimulation at 0.2 Hz (left), 0.5 Hz (center) and 1 Hz (right) in human ventricular cardiomyocytes incubated with scFv-hERG1 (10 μg/mL), in absence and presence of E4031 (5 µM). (D) Superimposed representative action potential traces recorded in human ventricular cardiomyocytes…

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  FIGURE 4. Comparison between the effect exerted by E4031 on action potential kinetics evaluated in ventricular cardiomyocytes isolated from patients with aortic stenosis (AOS) and incubated with scDB-hERG1-β1 or scFv-hERG1. (A) The figure shows the APD90 prolongation (%) promoted by the treatment with E4031 (5 µM) in ventricular cardiomyocytes incubated with scDB-hERG1-β1 (10 μg/mL) or scFv-hERG1 (10 μg/mL). The impact of E4031 on action potential kinetics is evaluated at different frequencies of stimulation: 0.2Hz, 0.5Hz, 1 Hz. * = p < 0.01; ** = p < 0.01; NS: not significant. (B–D) Action potential duration at 90 repolarization (APD90) recorded during stimulation at 0.2 Hz (B), 0.5 Hz (C) or 1 Hz (D) in human atrial cardiomyocytes incubated with hERG1-mAB and scFv-hERG1 (10 μg/mL). * = p < 0.05. Mean ± SE from 20 cardiomyocytes (10 patients). (E) Action potential duration at 90 repolarization (APD90) recorded during stimulation at 0.2 Hz in human ventricular cardiomyocytes incubated with hERG1-mAB and scFv-hERG1 (10 μg/mL). (F) Action potential duration at 90% repolarization (APD90) recorded during stimulation at 0.5 Hz in human ventricular cardiomyocytes incubated with hERG1-mAB and…

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  FIGURE 5. ECG trace and data analysis at time 0, 15, 30, 45, 60, 75, 90 min. (A) The ECG trace shows different time points 5 and 60 min after injection of the antibody. (B) Data analysis showing different parameters: RR interval (s), Heart Rate (BPM), PR interval (s), P Duration (s), QRS interval (s), QT interval (s), P Amplitude (V), Q Amplitude (V), R Amplitude (V), S Amplitude (V), T Amplitude (V).