AIMS: Human ether-a-go-go-related gene (hERG) missense mutations N588K and L532P are both associated with atrial fibrillation (AF). However, the underlying gain-of-function mechanism is different. The aim of this computational study is to assess and understand the arrhythmogenic mechanisms of these genetic disorders on the cellular and tissue level as a basis for the improvement of therapeutic strategies. METHODS AND RESULTS: The IKr formulation of an established model of human atrial myocytes was adapted by using the measurement data of wild-type and mutant hERG channels. Restitution curves of the action potential duration and its slope, effective refractory period (ERP), conduction velocity, reentry wavelength (WL), and the vulnerable window (VW) were determined in a one-dimensional (1D) tissue strand. Moreover, spiral wave inducibility and rotor lifetime in a 2D tissue patch were evaluated. The two mutations caused an increase in IKr regarding both peak amplitude and current integral, whereas the duration during which IKr is active was decreased. The WL was reduced due to a shorter ERP. Spiral waves could be initiated by using mutation models as opposed to the control case. The frequency dependency of the VW was reversed. CONCLUSION: Both mutations showed an increased arrhythmogenicity due to decreased refractory time in combination with a more linear repolarization phase. The effects were more pronounced for mutation L532P than for N588K. Furthermore, spiral waves presented higher stability and a more regular pattern for L532P. These in silico investigations unveiling differences of mutations affecting the same ion channel may help to advance genotype-guided AF prevention and therapy strategies.
Computational models of cardiac electrophysiology provided insights into arrhythmogenesis and paved the way toward tailored therapies in the last years. To fully leverage in silico models in future research, these models need to be adapted to reflect pathologies, genetic alterations, or pharmacological effects, however. A common approach is to leave the structure of established models unaltered and estimate the values of a set of parameters. Today's high-throughput patch clamp data acquisition methods require robust, unsupervised algorithms that estimate parameters both accurately and reliably. In this work, two classes of optimization approaches are evaluated: gradient-based trust-region-reflective and derivative-free particle swarm algorithms. Using synthetic input data and different ion current formulations from the Courtemanche et al. electrophysiological model of human atrial myocytes, we show that neither of the two schemes alone succeeds to meet all requirements. Sequential combination of the two algorithms did improve the performance to some extent but not satisfactorily. Thus, we propose a novel hybrid approach coupling the two algorithms in each iteration. This hybrid approach yielded very accurate estimates with minimal dependency on the initial guess using synthetic input data for which a ground truth parameter set exists. When applied to measured data, the hybrid approach yielded the best fit, again with minimal variation. Using the proposed algorithm, a single run is sufficient to estimate the parameters. The degree of superiority over the other investigated algorithms in terms of accuracy and robustness depended on the type of current. In contrast to the non-hybrid approaches, the proposed method proved to be optimal for data of arbitrary signal to noise ratio. The hybrid algorithm proposed in this work provides an important tool to integrate experimental data into computational models both accurately and robustly allowing to assess the often non-intuitive consequences of ion channel-level changes on higher levels of integration.
Inhibition of the atrial ultra-rapid delayed rectifier potassium current (I Kur) represents a promising therapeutic strategy in the therapy of atrial fibrillation. However, experimental and clinical data on the antiarrhythmic efficacy remain controversial. We tested the hypothesis that antiarrhythmic effects of I Kur inhibitors are dependent on kinetic properties of channel blockade. A mathematical description of I Kur blockade was introduced into Courtemanche-Ramirez-Nattel models of normal and remodeled atrial electrophysiology. Effects of five model compounds with different kinetic properties were analyzed. Although a reduction of dominant frequencies could be observed in two dimensional tissue simulations for all compounds, a reduction of spiral wave activity could be only be detected in two cases. We found that an increase of the percent area of refractory tissue due to a prolongation of the wavelength seems to be particularly important. By automatic tracking of spiral tip movement we find that increased refractoriness resulted in rotor extinction caused by an increased spiral-tip meandering. We show that antiarrhythmic effects of I Kur inhibitors are dependent on kinetic properties of blockade. We find that an increase of the percent area of refractory tissue is the underlying mechanism for an increased spiral-tip meandering, resulting in the extinction of re-entrant circuits.
A miniaturized ceramic atmospheric plasma source for the utilization in life sciences has been developed. It is manufactured in LTCC-technology (low temperature cofired ceramic). The plasma generation is based on buried electrodes which lead to a Dielectric Barrier Discharge (DBD). The employed technology allows small feature sizes (electrode width 150 μm, barrier thickness 40μm etc.) as well as precision in the μm range, resulting in a very low power consumption of the system (approx. 5 W). Thus, the maximum temperature at the point of use can be kept below 40 °C. The flexibility of the manufacturing process (layer lamination, screen printing, patterning with picosecond laser etc.) offers additional features like robust fluidic structures (channels, chambers, gas distribution etc.) as well as the direct implementation of electronic components. The technology concept as well as the design of the ceramic parts and the handhold matched to the multi-well plate format is demonstrated. The plasma of the system can be tuned depending on the assembly of the system and the electric excitation. To prove the biocompatibility and the experimental compatibility with cell cultures (low temperature at the point of use), a method for temperature measurements on the bottom of a multi-well plate was developed. First results of the impact of the plasma source on cell cultures are presented. The effects occurring in the plasma, as well as their effects on the cell cultures (ozone formation, ultraviolet radiation etc.) are separately considered. Furthermore, the cell tolerability of the treatment with the micro-plasma source is investigated with L929 fibroblast cells.
There is a large interest in analysing the QT-interval, as a prolonged QT-interval can cause the development of ventricular tachyarrhythmias such as Torsade de Pointes. One major part of QT-analysis is T-end detection. Three automatic T-end delineation methods based on wavelet fil- terbanks (WAM), correlation (CORM) and Principal Com- ponent Analysis PCA (PCAM) have been developed and applied to Physionet QT database. All algorithms tested on Physionet QT database showed good results, while PCAM produced better results than WAM and CORM achieved best results. Standard de- viation in sampling points (fs=250Hz) have been 33.3 (WAM), 8.0 (PTDM) and 7.8 (CORM). It could be shown that WAM is prone to interference while CORM is the most stable method even under bad conditions. Further- more it was possible to detect significant QT-prolongation caused by Moxifloxacin in Thorough QT Study # 2 us- ing CORM. QT-prolongation is significantly correlated to blood plasma concentration of Moxifloxacin.
Following the ICH E14 clinical evaluation guideline , the measurement of QT/QTc interval prolongation has become the standard surrogate biomarker for cardiac drug safety assessment and the faith of a drug development. In Thorough QT (TQT) study, a so-called positive control is employed to assess the ability of this study to detect the endpoint of interest, i.e. the QT prolongation by about five milliseconds. In other words the lower bound of the one- sided 95% confidence interval (CI) must be above 0 [ms]. Fully automated detection of ECG fiducial points and mea- surement of the corresponding intervals including QT in- tervals and RR intervals vary between different computer- ized algorithms. In this work we demonstrate the ability and reliability of Hannover ECG System (HES) to as- sess drug effects by detecting QT/QTc prolongation effects that meet the threshold of regulatory concern as mentioned by using THEW database studies namely TQT studies one and two.
While human ether-à-go-go-related gene (hERG) mutations N588K and K897T are associated with atrial fib- rillation (AF), the underlying arrhythmogenic mechanisms are understood only incompletely. In this work, an ap- proach integrating IKr measurement data from transgenic Xenopus oocytes into established computational models of cardiac electrophysiology is presented. Parameters are es- timated using a minimization formulation, which is handled by a hybrid particle swarm optimization (PSO) and trust- region-reflective (TRR) algorithm. Cell models adapted to the mutation measurements show a significantly shorter ac- tion potential (AP) with less pronounced spike-and-dome morphology. Results of single cell simulations compare with myocytes in chronic AF.