Analysis of the role of Nav1.5 slow inactivation in the development of inherited cardiac pathology

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Abstract

Voltage-gated cardiac sodium channels Nav1.5 are responsible for the initiation and propagation of action potentials in cardiomyocytes. Dysfunction of Nav1.5 can be caused both by pathogenic variants in the SCN5A gene itself, which encodes Nav1.5, and by genetic variants in the genes of other proteins, regulating channel activity and trafficking. The change of different phases of the action potential is determined by the strict temporal organization of activation and inactivation of various ion channels. Transitions between channel functional states (for example, to slow inactivated state) can be influenced by various factors and proteins interacting with the channel. Despite the fact that the process of slow inactivation of the channel has been known for several decades, its role in the mechanism of development of hereditary heart pathology remains unclear. In this work, using the patch clamp method in whole-cell leads, we studied changes in the process of slow Nav1.5 inactivation under the influence of various mutations in structural genes (DSP-H1684R, LMNA-R249Q, FLNC-R1267Q, FLNC-V2264M) associated with a genetically determined myocardial pathology leading to dysfunction of cardiomyocytes. The study used a model of cardiomyocytes differentiated from induced pluripotent stem cells (СM-iPSCs). We have demonstrated an increase in slow inactivation in the model of CM-iPSCs obtained from patients with a phenotype of cardiomyopathy combined with ventricular arrhythmias. Thus, this work contributes to understanding the role of the slow inactivation process in the mechanism of the development of heart pathology.

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A. K. Zaytseva

World-Class Research Centre for Personalized Medicine, Almazov National Medical Research Centre; Sechenov Institute of Evolutionary Physiology and Biochemistry of the Russian Academy of Sciences

Author for correspondence.
Email: zaytseva_ak@almazovcentre.ru
Russian Federation, Saint Petersburg; Saint Petersburg

K. I. Perepelina

World-Class Research Centre for Personalized Medicine, Almazov National Medical Research Centre; Institute of Cytology, Russian Academy of Sciences

Email: zaytseva_ak@almazovcentre.ru
Russian Federation, Saint Petersburg; Saint Petersburg

A. A. Kostareva

World-Class Research Centre for Personalized Medicine, Almazov National Medical Research Centre

Email: zaytseva_ak@almazovcentre.ru
Russian Federation, Saint Petersburg

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2. Fig. 1. Representative recordings of sodium currents registered in cells of the CM-IPSC model obtained from patients with hereditary heart pathology and healthy donors (donors 1, 2, 3). All represented lines generated INa of typical morphology

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3. Fig. 2. Enhancement of steady-state slow inactivation on the CM-IPSC model of a patient with progressive cardiac conduction disturbance and marked ventricular rhythm disturbances (DSP-H1684R, lower curve, n = 5) compared to the data obtained on cardiomyocytes from a healthy donor (upper curve, n = 9). * - differences are reliable at p < 0.05 (Mann-Whitney test). Here and in Figs. 3, 4 show mean values and their standard errors

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4. Fig. 3. Slow inactivation of Nav1.5 in BM-IPSCs derived from patients with Emery-Dreyfus dystrophy (LMNA-R249Q mutation, upper curve, n = 9) and in cells derived from a healthy donor (lower curve, n = 12). No changes in steady-state slow inactivation kinetics were detected at potentials from -80 to 20 mV (Mann-Whitney criterion)

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5. Fig. 4. Steady-state slow inactivation kinetics of Nav1.5 in cardiomyocytes of a patient with R1267Q mutation in the FLNC gene, V2264M mutation in the FLNC gene, and in cells from a healthy donor. The R1267Q mutation associated with a mixed phenotype (arrhythmogenic cardiomyopathy with severe ventricular arrhythmias) significantly altered the proportion of slowly inactivated channels (n = 6) compared with the current characteristics recorded in healthy donor cells (n = 12) at potentials ranging from -10 to 20 mV. The V2264M mutation in the FLNC gene associated with a mixed phenotype (restrictive cardiomyopathy, atrioventricular block, and ventricular tachycardia) did not alter the proportion of slowly inactivated channels (n = 7) relative to healthy donor cells. * - differences in the values of normalized current relative to healthy donors are reliable at p < 0.05 after correction (Mann-Whitney test with subsequent correction for multiple comparisons using the Benjamin-Hochberg method)

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