scholarly journals Transmural and rate-dependent profiling of drug-induced arrhythmogenic risks through in silico simulations of multichannel pharmacology

2019 ◽  
Author(s):  
Ping’an Zhao ◽  
Pan Li
Keyword(s):  
In Silico ◽  
Cell Types ◽  
M Cells ◽  
Cell Models ◽  
Drug Induced ◽  
Rate Dependent ◽  
Channel Inhibition ◽  
Inhibition Data ◽  
P Cells

AbstractBackgroundIn vitro hERG blockade assays alone provide insufficient information to accurately discriminate “safe” from “dangerous” drugs. Recent studies have suggested that the integration of multiple ion channel inhibition data can improve the prediction of drug-induced arrhythmogenic risks. In this study, using a family of cardiac cell models representing electrophysiological heterogeneities across the ventricular wall, we quantitatively evaluated transmural and rate-dependent properties of drug-induced arrhythmogenicity through computer simulations of multichannel pharmacology.Methods and ResultsRate-dependent drug effects of multiple ion channel inhibition on cardiac electrophysiology at their effective free therapeutic plasma concentrations (EFTPCs) were investigated using a group of in silico cell models (Purkinje (P) cells, endocardial (Endo) cells, mid-myocardial (M) cells and epicardial (Epi) cells). We found that (1) M cells are much more sensitive than the other cell types to drug-induced arrhythmias and can develop early afterdepolarization (EAD) in response to bepridil, dofetilide, sotalol, terfenadine, cisapride or ranolazine. (2) Most drug-induced adverse effects, such as pronounced action potential prolongations or EADs, occur at slower pacing rates. (3) Although most drug-induced EADs occur in M cells, the application of quinidine at its EFTPC can cause EADs in all four cell types. (4) The underlying ionic mechanism of drug-induced EADs differs across different cell types; while INaL is the major depolarizing current during the generation of EAD in P cells, ICaL is mostly predominant in other cell types. (5) Drug-induced AP alternans with larger beat-to-beat variations occur at high pacing rates in mostly P cells, while the application of bepridil can cause alternating EAD patterns at slower pacing rates in M cells.ConclusionsIn silico analysis of transmural and rate-dependent properties using multichannel inhibition data can be useful to accurately predict drug-induced arrhythmogenic risks and can also provide mechanistic insights into drug-induced adverse events related to cardiac arrhythmias.Author summaryIn vitro hERG blockade assays alone provide insufficient information to accurately discriminate “safe” from “dangerous” drugs, and computer simulation of ventricular action potential using multichannel inhibition data could be a useful tool to evaluate drug-induced arrhythmogenic risks. Our study suggested that the profiling of drug-induced transmural heterogeneities in cellular electrophysiology at all physiological pacing frequencies can be essential for the comprehensive evaluation of drug safety, and for the quantitative investigation into ionic mechanisms underlying drug-specific arrhythmogenic events. These in silico models and approaches may contribute to the ongoing construction of a comprehensive paradigm for the evaluation of drug-induced arrhythmogenic risks, potentially increase the success rate and accelerate the process of novel drug development.

scholarly journals Transmural and rate-dependent profiling of drug-induced arrhythmogenic risks through in silico simulations of multichannel pharmacology

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Ping’an Zhao ◽  
Pan Li
Keyword(s):  
Cardiac Electrophysiology ◽  
Cell Types ◽  
Cell Models ◽  
Insufficient Information ◽  
Drug Induced ◽  
Rate Dependent ◽  
Early Afterdepolarization ◽  
P Cells ◽  
Quantitative Profiling

AbstractIn vitro human ether-à-go-go related gene (hERG) inhibition assay alone might provide insufficient information to discriminate “safe” from “dangerous” drugs. Here, effects of multichannel inhibition on cardiac electrophysiology were investigated using a family of cardiac cell models (Purkinje (P), endocardial (Endo), mid-myocardial (M) and epicardial (Epi)). We found that: (1) QT prolongation alone might not necessarily lead to early afterdepolarization (EAD) events, and it might be insufficient to predict arrhythmogenic liability; (2) the occurrence and onset of EAD events could be a candidate biomarker of drug-induced arrhythmogenicity; (3) M cells are more vulnerable to drug-induced arrhythmias, and can develop early afterdepolarization (EAD) at slower pacing rates; (4) the application of quinidine can cause EADs in all cell types, while INaL is the major depolarizing current during the generation of drug-induced EAD in P cells, ICaL is mostly responsible in other cell types; (5) drug-induced action potential (AP) alternans with beat-to-beat variations occur at high pacing rates in P cells. These results suggested that quantitative profiling of transmural and rate-dependent properties can be essential to evaluate drug-induced arrhythmogenic risks, and may provide mechanistic insights into drug-induced arrhythmias.

scholarly journals Urine-Derived Epithelial Cells as Models for Genetic Kidney Diseases

Cells ◽  
2021 ◽  
Vol 10 (6) ◽  
pp. 1413
Author(s):  
Tjessa Bondue ◽  
Fanny O. Arcolino ◽  
Koenraad R. P. Veys ◽  
Oyindamola C. Adebayo ◽  
Elena Levtchenko ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Kidney Diseases ◽  
Cell Types ◽  
Cell Models ◽  
Tubular Epithelial Cells ◽  
Disease Mechanisms ◽  
Proximal Tubular Epithelial Cells ◽  
Proximal Tubular ◽  
Disease Cell

Epithelial cells exfoliated in human urine can include cells anywhere from the urinary tract and kidneys; however, podocytes and proximal tubular epithelial cells (PTECs) are by far the most relevant cell types for the study of genetic kidney diseases. When maintained in vitro, they have been proven extremely valuable for discovering disease mechanisms and for the development of new therapies. Furthermore, cultured patient cells can individually represent their human sources and their specific variants for personalized medicine studies, which are recently gaining much interest. In this review, we summarize the methodology for establishing human podocyte and PTEC cell lines from urine and highlight their importance as kidney disease cell models. We explore the well-established and recent techniques of cell isolation, quantification, immortalization and characterization, and we describe their current and future applications.

scholarly journals Trichostatin A downregulates bromodomain and extra-terminal proteins to suppress osimertinib resistant non-small cell lung carcinoma

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Yuting Meng ◽  
Xixi Qian ◽  
Li Zhao ◽  
Nan Li ◽  
Shengjie Wu ◽  
...  
Keyword(s):  
Trichostatin A ◽  
Small Cell Lung Carcinoma ◽  
Cell Types ◽  
Inhibitory Effects ◽  
Cell Lung Carcinoma ◽  
Growth Inhibitory ◽  
Terminal Proteins ◽  
P Cells

Abstract Background The third-generation epithelial growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs) have shown significant therapeutic effects on patients with non-small cell lung carcinoma (NSCLC) who carry active EGFR mutations, as well as those who have developed acquired resistance to the first-generation of EGFR-TKIs due to the T790M mutation. However, most patients develop drug resistance after 8–10 months of treatment. Currently, the mechanism has not been well clarified, and new therapeutic strategies are urgently needed. Methods Osimertinib resistant cell lines were established by culturing sensitive cells in chronically increasing doses of osimertinib. The anticancer effect of reagents was examined both in vitro and in vivo using the sulforhodamine B assay and a xenograft mouse model. The molecular signals were detected by western blotting. The combination effect was analyzed using CompuSyn software. Results We found that bromodomain and extra-terminal proteins (BETs) were upregulated in osimertinib resistant (H1975-OR) cells compared with those in the paired parental cells (H1975-P), and that knockdown of BETs significantly inhibited the growth of H1975-OR cells. The BET inhibitor JQ1 also exhibited stronger growth-inhibitory effects on H1975-OR cells and a greater expression of BETs and the downstream effector c-Myc than were observed in H1975-P cells. The histone deacetylase (HDAC) inhibitor trichostatin A (TSA) showed stronger growth suppression in H1975-OR cells than in H1975-P cells, but vorinostat, another HDAC inhibitor, showed equal inhibitory efficacy in both cell types. Consistently, downregulation of BET and c-Myc expression was greater with TSA than with vorinostat. TSA restrained the growth of H1975-OR and H1975-P xenograft tumors. The combination of TSA and JQ1 showed synergistic growth-inhibitory effects in parallel with decreased BET and c-Myc expression in both H1975-OR and H1975-P cells and in xenograft nude mouse models. BETs were not upregulated in osimertinib resistant HCC827 cells compared with parental cells, while TSA and vorinostat exhibited equal inhibitory effects on both cell types. Conclusion Upregulation of BETs contributed to the osimertinib resistance of H1975 cells. TSA downregulated BET expression and enhanced the growth inhibitory effect of JQ1 both in vitro and in vivo. Our findings provided new strategies for the treatment of osimertinib resistance.

scholarly journals Drug-Induced Liver Injury and Individual Cell Models

2015 ◽  
Vol 33 (4) ◽  
pp. 486-491 ◽  
Author(s):  
Andreas Benesic ◽  
Alexander L. Gerbes
Keyword(s):  
Liver Injury ◽  
Individual Cell ◽  
Clinical Situation ◽  
Clinical Development ◽  
Cell Models ◽  
Drug Induced ◽  
In Vitro Susceptibility ◽  
Drug Induced Liver Injury ◽  
The Individual

Drug-induced liver injury (DILI) is the most common cause of acute liver failure and accounts for the majority of regulatory actions on drugs. Furthermore, DILI is a relevant cause for project terminations in pharmaceutical development. The idiosyncratic form of DILI is especially a threat in late clinical development phases and postmarketing, respectively. Even the occurrence of only a few idiosyncratic DILI cases in late clinical development or postmarketing may suffice to terminate or withdraw an otherwise promising therapy. Despite advances in preclinical assessment of dose-dependent toxicity, idiosyncratic DILI is still a big challenge for in vitro research: it not only requires individualized models but also a huge number of tests. We have developed and investigated MetaHeps®, a technology involving hepatocyte-like cells generated from peripheral monocytes without genetic modifications. These cells exhibit several hepatocyte-like characteristics and show donor-specific activities of drug-metabolizing enzymes. With MetaHeps we have performed in vitro investigations in patients with DILI suspicion. By investigating MetaHeps derived from DILI patients we could show increased in vitro susceptibility to the drugs involved in the individual patients. MetaHeps testing could also rule out DILI and help to identify other causes of acute liver injury. Moreover, MetaHeps identified the causative agent in polymedicated patients. In conclusion, in vitro research of idiosyncratic DILI requires individual cell models which produce results comparable to the clinical situation. We suggest the MetaHeps technology as a novel tool to cope with these challenges of DILI.

scholarly journals In-silico clinical trial using high performance computational modeling of a virtual human cardiac population to assess drug-induced arrhythmic risk

2021 ◽  
Author(s):  
Jazmin Aguado-Sierra ◽  
Constantine Butakoff ◽  
Renee Brigham ◽  
Apollo Baron ◽  
Guillaume Houzeaux ◽  
...  
Keyword(s):  
Clinical Trial ◽  
In Silico ◽  
High Performance ◽  
Drug Induced ◽  
Computational Framework ◽  
Major Health ◽  
Action Potential Prolongation ◽  
Performance Computing ◽  
Major Health Issue

AbstractDrug-induced arrhythmia continues to be a major health issue worldwide. The need for reliable pro-arrhythmic predictors became relevant during early phases of the SarsCoV2 pandemic, when it was uncertain whether the use of hydroxychloroquine (HCQ) and azithromycin (AZM) could be more harmful than beneficial due to their reported pro-arrhythmic effects.In this work we describe a computational framework that employs a gender-specific, in-silico cardiac population to assess cardiac drug-induced QT-prolongation after the administration of a single or a combination of potentially cardiotoxic drugs as HCQ and AZM. This novel computational methodology is capable of reproducing the complex behavior of the clinical electrocardiographic response to drug-induced arrhythmic risk, in-silico. Using high performance computing, the computational framework allows the estimation of the arrhythmic risk in a population, given a variety of doses of one or more drugs in a timely manner and providing markers that can be directly related to the clinical scenario. The pro-arrhythmic behavior observed in subjects within the in-silico trial, was also compared to supplemental in-vitro experiments on a reanimated swine hearts. Evidence of transmurally heterogeneous action potential prolongation after the administration of a large dose of HCQ was an observed mechanism of arrhythmia, both in the in-vitro and the in-silico model. The virtual clinical trial also provided remarkably similar results to recent published clinical data. In conclusion, the in-silico clinical trial on the cardiac population is capable of reproducing and providing evidence of the normal phenotype variants that produce distinct arrhythmogenic outcomes after the administration of one or various drugs.

scholarly journals A Critical Perspective on 3D Liver Models for Drug Metabolism and Toxicology Studies

2021 ◽  
Vol 9 ◽  
Author(s):  
Ana S. Serras ◽  
Joana S. Rodrigues ◽  
Madalena Cipriano ◽  
Armanda V. Rodrigues ◽  
Nuno G. Oliveira ◽  
...  
Keyword(s):  
Drug Development ◽  
Liver Toxicity ◽  
Three Dimensional ◽  
Cell Types ◽  
Cell Phenotype ◽  
Drug Induced ◽  
Characterization Methods ◽  
Clinical Phase ◽  
Post Marketing

The poor predictability of human liver toxicity is still causing high attrition rates of drug candidates in the pharmaceutical industry at the non-clinical, clinical, and post-marketing authorization stages. This is in part caused by animal models that fail to predict various human adverse drug reactions (ADRs), resulting in undetected hepatotoxicity at the non-clinical phase of drug development. In an effort to increase the prediction of human hepatotoxicity, different approaches to enhance the physiological relevance of hepatic in vitro systems are being pursued. Three-dimensional (3D) or microfluidic technologies allow to better recapitulate hepatocyte organization and cell-matrix contacts, to include additional cell types, to incorporate fluid flow and to create gradients of oxygen and nutrients, which have led to improved differentiated cell phenotype and functionality. This comprehensive review addresses the drug-induced hepatotoxicity mechanisms and the currently available 3D liver in vitro models, their characteristics, as well as their advantages and limitations for human hepatotoxicity assessment. In addition, since toxic responses are greatly dependent on the culture model, a comparative analysis of the toxicity studies performed using two-dimensional (2D) and 3D in vitro strategies with recognized hepatotoxic compounds, such as paracetamol, diclofenac, and troglitazone is performed, further highlighting the need for harmonization of the respective characterization methods. Finally, taking a step forward, we propose a roadmap for the assessment of drugs hepatotoxicity based on fully characterized fit-for-purpose in vitro models, taking advantage of the best of each model, which will ultimately contribute to more informed decision-making in the drug development and risk assessment fields.

scholarly journals Kidney-based in vitro models for drug-induced toxicity testing

2019 ◽  
Vol 93 (12) ◽  
pp. 3397-3418 ◽  
Author(s):  
João Faria ◽  
Sabbir Ahmed ◽  
Karin G. F. Gerritsen ◽  
Silvia M. Mihaila ◽  
Rosalinde Masereeuw
Keyword(s):  
Adverse Effects ◽  
Drug Disposition ◽  
Drug Effects ◽  
Toxicity Testing ◽  
Cell Types ◽  
In Vitro Assays ◽  
Vascular Effects ◽  
Drug Induced

Abstract The kidney is frequently involved in adverse effects caused by exposure to foreign compounds, including drugs. An early prediction of those effects is crucial for allowing novel, safe drugs entering the market. Yet, in current pharmacotherapy, drug-induced nephrotoxicity accounts for up to 25% of the reported serious adverse effects, of which one-third is attributed to antimicrobials use. Adverse drug effects can be due to direct toxicity, for instance as a result of kidney-specific determinants, or indirectly by, e.g., vascular effects or crystals deposition. Currently used in vitro assays do not adequately predict in vivo observed effects, predominantly due to an inadequate preservation of the organs’ microenvironment in the models applied. The kidney is highly complex, composed of a filter unit and a tubular segment, together containing over 20 different cell types. The tubular epithelium is highly polarized, and the maintenance of this polarity is critical for optimal functioning and response to environmental signals. Cell polarity is dependent on communication between cells, which includes paracrine and autocrine signals, as well as biomechanic and chemotactic processes. These processes all influence kidney cell proliferation, migration, and differentiation. For drug disposition studies, this microenvironment is essential for prediction of toxic responses. This review provides an overview of drug-induced injuries to the kidney, details on relevant and translational biomarkers, and advances in 3D cultures of human renal cells, including organoids and kidney-on-a-chip platforms.

From in vivo plasma composition to in vitro cardiac electrophysiology and in silico virtual heart: the extracellular calcium enigma

2009 ◽  
Vol 367 (1896) ◽  
pp. 2203-2223 ◽  
Author(s):  
Stefano Severi ◽  
Cristiana Corsi ◽  
Elisabetta Cerbai
Keyword(s):  
In Silico ◽  
Cardiac Electrophysiology ◽  
Cell Models ◽  
Laboratory Measurements ◽  
Cardiac Action ◽  
Potential Impact ◽  
The Difference ◽  
Physiological Values

In spite of its potential impact on simulation results, the problem of setting the appropriate Ca 2+ concentration ([Ca 2+ ] o ) in computational cardiac models has not yet been properly considered. Usually [Ca 2+ ] o values are derived from in vitro electrophysiology. Unfortunately, [Ca 2+ ] o in the experiments is set significantly far (1.8 or 2 mM) from the physiological [Ca 2+ ] in blood (1.0–1.3 mM). We analysed the inconsistency of [Ca 2+ ] o among in vivo , in vitro and in silico studies and the dependence of cardiac action potential (AP) duration (APD) on [Ca 2+ ] o . Laboratory measurements confirmed the difference between standard extracellular solutions and normal blood [Ca 2+ ]. Experimental data on human atrial cardiomyocytes confirmed literature data, demonstrating an inverse relationship between APD and [Ca 2+ ] o . Sensitivity analysis of APD on [Ca 2+ ] o for five of the most used cardiac cell models was performed. Most of the models responded with AP prolongation to increases in [Ca 2+ ] o , i.e. opposite to the AP shortening observed in vitro and in vivo. Modifications to the Ten Tusscher–Panfilov model were implemented to demonstrate that qualitative consistency among in vivo , in vitro and in silico studies can be achieved. The Courtemanche atrial model was used to test the effect of changing [Ca 2+ ] o on quantitative predictions about the effect of K + current blockade. The present analysis suggests that (i) [Ca 2+ ] o in cardiac AP models should be changed from 1.8 to 2 mM to approximately 1.15 mM in order to reproduce in vivo conditions, (ii) the sensitivity to [Ca 2+ ] o of ventricular AP models should be improved in order to simulate real conditions, (iii) modifications to the formulation of Ca 2+ -dependent I CaL inactivation can make models more suitable to analyse AP when [Ca 2+ ] o is set to lower physiological values, and (iv) it could be misleading to use non-physiological high [Ca 2+ ] o when the quantitative analysis of in vivo pathophysiological mechanisms is the ultimate aim of simulation.

scholarly journals Ribavirin shows antiviral activity against SARS-CoV-2 and downregulates the activity of TMPRSS2 and the expression of ACE2 in vitro

2021 ◽  
pp. 1-12 ◽  
Author(s):  
Mehmet Altay Unal ◽  
Ceylan Verda Bitirim ◽  
Gokce Yagmur Summak ◽  
Sidar Bereketoglu ◽  
Inci Cevher Zeytin ◽  
...  
Keyword(s):  
Antiviral Activity ◽  
Broad Spectrum ◽  
In Silico ◽  
Antiviral Drug ◽  
In Silico Analysis ◽  
Cell Types ◽  
Suppressive Effect ◽  
Inhibitory Effect ◽  
Molecular Techniques

Ribavirin is a guanosine analog with broad-spectrum antiviral activity against RNA viruses. Based on this, we aimed to show the anti-SARS-CoV-2 activity of this drug molecule via in vitro, in silico, and molecular techniques. Ribavirin showed antiviral activity in Vero E6 cells following SARS-CoV-2 infection, whereas the drug itself did not show any toxic effect over the concentration range tested. In silico analysis suggested that ribavirin has a broad-spectrum impact on SARS-CoV-2, acting at different viral proteins. According to the detailed molecular techniques, ribavirin was shown to decrease the expression of TMPRSS2 at both mRNA and protein levels 48 h after treatment. The suppressive effect of ribavirin in ACE2 protein expression was shown to be dependent on cell types. Finally, proteolytic activity assays showed that ribavirin also showed an inhibitory effect on the TMPRSS2 enzyme. Based on these results, we hypothesized that ribavirin may inhibit the expression of TMPRSS2 by modulating the formation of inhibitory G-quadruplex structures at the TMPRSS2 promoter. As a conclusion, ribavirin is a potential antiviral drug for the treatment against SARS-CoV-2, and it interferes with the effects of TMPRSS2 and ACE2 expression.

scholarly journals Comparison of the Simulated Response of Three in Silico Human Stem Cell-Derived Cardiomyocytes Models and in Vitro Data Under 15 Drug Actions

2021 ◽  
Vol 12 ◽  
Author(s):  
Michelangelo Paci ◽  
Jussi T. Koivumäki ◽  
Hua Rong Lu ◽  
David J. Gallacher ◽  
Elisa Passini ◽  
...  
Keyword(s):  
Experimental Data ◽  
Stem Cell ◽  
In Silico ◽  
Drug Testing ◽  
Drug Application ◽  
Human Stem Cell ◽  
Drug Induced ◽  
Drug Actions ◽  
Experimental Findings

Objectives: Improvements in human stem cell-derived cardiomyocyte (hSC-CM) technology have promoted their use for drug testing and disease investigations. Several in silico hSC-CM models have been proposed to augment interpretation of experimental findings through simulations. This work aims to assess the response of three hSC-CM in silico models (Koivumäki2018, Kernik2019, and Paci2020) to simulated drug action, and compare simulation results against in vitro data for 15 drugs.Methods: First, simulations were conducted considering 15 drugs, using a simple pore-block model and experimental data for seven ion channels. Similarities and differences were analyzed in the in silico responses of the three models to drugs, in terms of Ca2+ transient duration (CTD90) and occurrence of arrhythmic events. Then, the sensitivity of each model to different degrees of blockage of Na+ (INa), L-type Ca2+ (ICaL), and rapid delayed rectifying K+ (IKr) currents was quantified. Finally, we compared the drug-induced effects on CTD90 against the corresponding in vitro experiments.Results: The observed CTD90 changes were overall consistent among the in silico models, all three showing changes of smaller magnitudes compared to the ones measured in vitro. For example, sparfloxacin 10 µM induced +42% CTD90 prolongation in vitro, and +17% (Koivumäki2018), +6% (Kernik2019), and +9% (Paci2020) in silico. Different arrhythmic events were observed following drug application, mainly for drugs affecting IKr. Paci2020 and Kernik2019 showed only repolarization failure, while Koivumäki2018 also displayed early and delayed afterdepolarizations. The spontaneous activity was suppressed by Na+ blockers and by drugs with similar effects on ICaL and IKr in Koivumäki2018 and Paci2020, while only by strong ICaL blockers, e.g. nisoldipine, in Kernik2019. These results were confirmed by the sensitivity analysis.Conclusion: To conclude, The CTD90 changes observed in silico are qualitatively consistent with our in vitro data, although our simulations show differences in drug responses across the hSC-CM models, which could stem from variability in the experimental data used in their construction.

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