CDI welcomes you to join us at Applied Pharmaceutical Toxicology where we will be presenting the latest on toxicity testing with our iCell and MyCell products.
- Talk: Solutions to Improve Toxicity Testing Relevance and Predictivity
- Poster: Evaluating Networked Activity as an Integrated Assay for Seizurogenic Assessment Using Stem Cell Derived Glutamatergic Neurons
- Poster: 3D and 2D in Vitro Models of Xenobiotic-Induced Hepatotoxicity using iCell® Hepatocytes 2.0
- Poster: Advances in Human Induced Pluripotent Stem Cell Derived Cardiomyocytes: Maintaining Validated Responses and Enabling Relevant Toxicity Testing
CDI Activity at Applied Pharmaceutical Toxicology 2017
Cellular Dynamics Podium Presentation
Tuesday, May 16 | 1:30 – 2 pm
Human iPSC-Derived Cardiomyocytes, Glutamatergic Neurons, and Hepatocytes; Cells and Solutions to Improve Toxicity Testing Relevance and Predictivity
Blake Anson, PhD, Cellular Dynamics International
Cellular Dynamics Poster Presentations
Evaluating Networked Activity as an Integrated Assay for Seizurogenic Assessment Using Stem Cell-derived Glutamatergic Neurons
Kwi Hye Kim, Coby Carlson, Christian Kannemeier, Tromondae K Feaster, Brad Swanson, Blake Anson and Kile Mangan
Cellular Dynamics International
Some ‘regular’ prescription medications are known to display seizurogenic potential and these adverse events can be mis-diagnosed as epilepsy. Therefore it is crucial to test for seizurogenic potential during drug development. Central to this need is the requirement for human material and the potential utility of human induced pluripotent stem cell (iPSC) technology. Here we present human iPSC-derived excitatory neuronal populations (e.g. iCell® GlutaNeurons) that develop and display network-level coordinated spontaneous activity in vitro as evidenced by synchronized bursts captured and measured via MEA. This technology provides a unique means to assess and alter if necessary, early in the drug discovery process the propensity for compound-mediate modulation of neuronal electrical activity.
Excitatory human neurons (iCell Glutaneurons) were derived using induced pluripotent stem cell technology. Spontaneous electrical activity, including synchronized bursting, was measured via micro-electrode array (MEA) technology before and after reference compound application.
Excitatory populations of iPSC-derived cortical neurons (i.e., iCell GlutaNeurons) develop and display network-level coordinated neuronal activity in vitro, evident by synchronized bursts captured and measured via MEA. Assay optimization dictates best practice timelines for ‘seizurogenic potential’ screening occurs between DIV20-23, post-thaw. Excitatory pharmacology that displays concentration-dependent seizurogenic-effects include picrotoxin [0.3-100 µM], GABAzine [1-100 µM], bicuculline [4-400 µM], pentylenetetrazol [7 µM-2 mM], 4-aminopyridine [1.6-50 µM], and kainic acid [0.4-300 µM]. Activity metrics displaying concentration-dependent changes with pharmacology include: mean firing rate, ‘single-channel’ burst rate, intensity and duration, ‘network-level’ burst rate, intensity and duration, and synchrony measures.
These data establish human iPSC-derived GlutaNeurons as a reliable and predictive model. The presented data illustrate and couple the “seizure-in-a-dish” technology, previously limited to rodent-only investigation, with human iPSC-derived neurons to create an unprecedented investigatory space for assessing seizurogenic risk potential.
Hepatotoxicity is a leading cause of drug withdrawal from the market, and current preclinical models are not sufficiently predictive of drug effects in humans. Causes of hepatotoxicity include intrinsic toxic effects and the enzymatic production of toxic metabolites. Development of more predictive in vitro model systems to identify hepatotoxicity early in drug development is critical for decision making and to avoid Drug Induced Liver Injury in the clinic. Moreover, batch to batch and donor inconsistencies in primary human hepatocytes, as well as lack of maintained metabolic function have resulted in conflicting reports and poor predicticity.
Here, we set out to demonstrate the functional utility of iCell Hepatocytes 2.0 (HC 2.0) to assess acute and chronic drug-induced hepatotoxicity. Human induced pluripotent stem cell (iPSC)-derived hepatocytes (iCell® Hepatocytes 2.0) exhibit high purity and sustained biologically relevant functions help to address some of the needs of hepatotoxicity assessment.
We evaluated HC 2.0 responses to a set of known hepatotoxins (i.e. amiodarone, acetaminophen, troglitazone, nefazadone, chlorpromazine, and FCCP) across a number of cell death readouts highlighting their capacity for mechanistic toxicity studies. In addition, the prolonged viability also enables chronic dosing in vitro affording the potential to detect the effects of slow to form metabolites and also perform analyses at physiologically relevant concentrations over protracted exposure periods. The short term high concentration sensitivities observed were comparable to those seen with primary human hepatocytes. However, effects seen over 48 hr and 7 day dosing are illustrative of the potential of HC 2.0 for predictive in vivo/in vitro toxicity correlation. With the ability to routinely access patient specific genotypes and also culture in 3D spheroids and in co-culture with other hepatic stellate cells HC 2.0 provide a biologically relevant human model system for investigating hepatotoxicity in preclinical drug development.
These data illustrate how human-based iCell products offer an excellent model system for assessing compound effects in human-derived hepatocytes. In total, iPSC technology enables a reliable and predictive model systems not previously attainable, and provides new solutions, tools, and opportunities for more predictive toxicity testing.
Advances in Human Induced Pluripotent Stem Cell Derived Cardiomyocytes: Maintaining Validated Responses and Enabling Relevant Toxicity Testing
The combined efforts of academic, pharmaceutical, and regulatory scientists in the large-scale Comprehensive In-Vitro Proarrhythmia assay (CiPA), Japan iPS Cardiac Safety Assessment (JiCSA), and Consortium for Safety Assessment using Human iPS Cells (CSAHi) consortia demonstrates the prominent role of human induced pluripotent stem cell (hiPSC)- cardiomyocytes across all sectors of research. As hiPSC-cardiomyocytes continue to evolve, it is imperative to understand their performance in relation to previously validated models and versions.
This study used the CiPA compound collection and the multi-electrode array (MEA) platform to evaluate the responses of two generations of commercially available hiPSC-derived cardiomyocytes, iCell Cardiomyocytes2 and iCell Cardiomyocytes.
Baseline data demonstrated stable beating rates for both iCell Cardiomyocytes2 and iCell Cardiomyocytes. The single channel blocking compounds mexilitine, nifedipine, E-4031, and JNJ-303 and the multi-channel blocking compounds flecainide, moxifloxacin, quinidine, and ranolazine produced quantifiable effects on cellular electrophysiology that were comparable across both cell types and culture conditions.
The concordance of the data indicate that despite negligible changes in beat rate, iCell Cardiomyocytes and iCell Cardiomyocytes2 respond similarly in chemical space and thus bridging studies should be sufficient for adopting new product iterations.