24.09.2017 | HTS in Cardiac Safety
Conference Venue: Maritim Hotel Berlin, Germany
Niels Fertig, Sonja Stölzle-Feix, Frank Henrichsen and Elena Dragicevic are looking forward to meeting you on Sunday 24th September, 4:00 PM - 6:30 PM in the Maritim Hotel Salon 15 / Paris (1st floor).
Said el Haou, Metrion Biosciences:
„In depth profiling of human iPSC-CM – from electrophysiology to phenotypic assays"
Matt Burnham, AstraZeneca:
„Impedance and field potential in 3D: cardiomyocyte co-culture spheroids and the CardioExcyte96 platform"
In vitro three-dimensional and co-culture cell models can provide more physiologically relevant data than conventional 2D approaches. We explored novel label-free methods to record functional data from actively beating, 3D spheroid co-cultures of iPSC cardiomyocytes. Using the Nanion CardioExcyte® 96 platform, designed for recordings from 2D iPSC cardiomyocytes monolayers, the electrical activity (extracellular field potential) and beating pattern (impedance) of cardiomyocyte spheroids could be detected. To facilitate the essential spheroid-to-electrode positioning, magnetic nano-particle based approaches were explored. Treatment with compounds known to affect beat rate and field potential duration generated appropriate results in the 3D system. A cumulative-dosing approach was developed to demonstrate a drug discovery work flow suitable for profiling studies. In comparison to a conventional contractility assay approach (2D FLIPR), significant advantages in reduced iPSC usage and cost (approximately eight-fold) and increased through-put were demonstrated. The high through-put of the system was leveraged to profile the functional phenotype of various iPSC and fibroblast co-culture types under different experimental conditions. Phenotype profiling revealed substantial differences amongst co-culture types, such as in beat amplitude and responses to E-4031. Electrical pacing revealed true inotropic responses (increased beat amplitude at constant beat rate) in some conditions. In summary, recording functional data from the 3D phenotype of spheroid co-cultures of electrically-active and beating cardiomyocytes in a screening platform has been achieved which may improve the predictivity of Safety assays and facilitate a better understanding of 3D co-culture phenotypes.
Takasuna Kiyoshi, Daiichi Sankyo/ Novare:
„Comprehensive in vitro cardiac safety assessment using human stem cell technology (CSAHi: HEART initiative) -The second stage"
1) Discovery Pharmacokinetics Research Group, Pharmaceutical and Biomedical Analysis Department, Daiichi Sankyo Rd Novare Co., Ltd.
2) Consortium for Safety Assessment using Human iPS Cells (CSAHi): HEART Team
Since 2013, we have organized the Consortium for Safety Assessment using Human iPS Cells (CSAHi; http://csahi.org/en/) in Japan to verify the application of human iPS/ES cell-derived cardiomyocytes in drug safety evaluation. The definitive goal of the CSAHi HEART Team has been to propose comprehensive screening strategies to predict a diverse range of cardiotoxicities (arrhythmia, contractile dysfunction, and cardiomyocyte toxicity) by using recently introduced platforms (multi-electrode array [MEA], patch clamp, cellular impedance, motion field imaging [MFI], and Ca transient systems) while identifying the strengths and weaknesses of each.
Our study (the first stage) showed that hiPS-CMs used in these platforms would offer paradigm changes of platforms for predicting drug-induced QT risk and/or arrhythmia or contractile dysfunction with a greater prognostic value beyond existing indirect surrogates proposed in animal models. I will introduce the latest progress and roadmap of activities of the CSAHi HEART Team (the second stage).
Teun de Boer, University of Utrecht:
„Dynamic clamping on a Patchliner: adding virtual IK1 channels to cardiomyocytes"
Department of Medical Physiology, Division Heart & Lungs, University Medical Center Utrecht. Yalelaan 50, 3584 CM Utrecht, The Netherlands
Human pluripotent stem cell-derived cardiomyocytes (hPSC-CM) are an interesting source of cells for safety pharmacology, but there are caveats that have to be taken into account. A typical property of hPSC-CM is automaticity, the cardiomyocytes beat spontaneously, resembling cells in the sinoatrial node of the heart. The spontaneous beating is due to lack of IK1 ion channels, which normally assure a stable resting membrane potential in cardiomyocytes. Work by our group has demonstrated that the depolarized state of hPSC-CM limits their usefulness in assays aimed at detecting proarrhythmic properties of drugs (Jonsson et al., 2012, PMID: 22353256).
Dynamic clamping can provide virtual, simulated IK1 channels to a real biological hPSC-CM in a patch clamping experiment. Key benefits of this approach include full control of the added IK1 conductance to each cardiomyocyte, it can be applied to any hPSC-CM source and it can be coupled to automated patch clamping machines. In this webinar I will discuss our work on using the dynamic clamping technique with a Patchliner.
The symposium is free to attend.
Download the program here.