Developing in vitro based alternatives to animal testing

Speakers: Dr. Ivan Kopljar, Dr. Peter Racz, Dr. Andy Forreryd
Chair: Prof. Juliette Legler

Animal experimentation has traditionally played a vital role in both the discovery and safety assessment of clinical compounds. However, in recent years the issue of animal testing has become increasingly controversial in the public eye, adding momentum to the existing drive to develop animal-free models that accurately reflect toxic effects in humans. Today we are going to discuss some recent progress towards this goal as we hear about three state-of-the-art in vitro based systems for toxicological testing. The session will begin with a brief introduction followed by a presentation by Dr Ivan Kopljar on the development of human stem cell-derived cardiomyocytes for cardiac safety assessment. In the second presentation Dr Peter Racz will discuss the development of a stem cell-based reporter assay for developmental and reproductive toxicology. The third speaker Dr Andy Forreryd will present a in vitro testing platform for assessing skin and respiratory sensitization genomic based probes. This session will be concluded with poster pitches relative to the topic will be given by young scientists.

 

Human stem cell-derived cardiomyocytes in cardiac safety assessment: Insides into Janssen’s strategy towards early hazard identification.

Ivan Kopljar

Cardiac safety assessment of new chemical entities (NCEs) is an important pillar within drug discovery and preclinical development. Current approaches provide sufficient indications of risk but require various in vitro and in vivo models involving multiple animal species. The need for more human-based cardiac models together with the effort to implement the 3R’s (Replace, Reduce, Refine) concept with respect to preclinical animal use, has put the spotlight on human induced pluripotent stem cell derived cardiomyocytes (hiPSC-CMs). This human-based model has been extensively evaluated over the recent years and is currently being applied within the pharmaceutical industry. Here, we will discuss Janssen’s approach to validate and introduce hiPSC-CMs-based screening assays for early preclinical cardiac safety evaluation of NCEs and demonstrate the positive implications on the 3R’s concept of animal use.

 

Validation of a Novel Human Stem Cell-Based Gene Expression Assay for In Vitro DART Assessment.

Peter Racz

Testing for developmental and reproductive toxicology (DART) is a crucial part of the toxicological risk assessment. Today, DART mostly relies on animal testing, although alternative in vitro tests are being used. However, these in vitro assays often do not provide mechanistic insight, and the results are difficult to translate to human risk due to inter-species differences. To improve the in vitro identification of developmental toxicants, we developed an assay in which we followed the differentiation of human induced pluripotent stem cells (hiPSC) towards cardiomyocytes, hepatocytes or neural rosettes by measuring the expression of various biomarker genes. Changes in biomarker expression can be used to identify disruption of the tightly timed and regulated early development program.

 

In vitro testing for skin and respiratory sensitizers using a genomic biomarker signature and machine learning

Andy Forreryd

Exposure to chemicals with sensitizing properties may result in priming of allergic hypersensitivity reactions in either the skin or the respiratory tract. In recent years, efforts have been made to develop predictive assays for prospective identification of compounds capable of inducing such reactions. To date, several assays for assessment of skin sensitizers have been proposed, but the need for an assay that specifically identifies chemical respiratory sensitizers remains unfulfilled. The absence of predictive methods for chemical respiratory sensitizers is partly due to a lack of understanding of the exact immunobiological mechanisms giving rise to the toxicological outcome. However, a key mechanistic difference is represented by a preference of skin or respiratory sensitizers to induce qualitatively different T-cell populations, which in turn is assumed to be dependent on differential signaling from dendritic cells (DCs). Subsequently, by looking for unique mechanistic differences in DCs, it should be possible to not only identify potential respiratory sensitizers, but also distinguish them from skin sensitizers. This talk describes the development of the Genomic Allergen Rapid Detection (GARD) platform and describes its applicability for testing of both skin and respiratory sensitizing chemicals. The assay is based on a human DC-like cell line and uses state-of-the-art machine learning algorithms to classify unknown test substances based on the readout from genomic biomarker signatures of genes involved in immunologically relevant pathways. The talk will demonstrate on the benefits of embracing novel transcriptomic technologies, emerging computational tools and powerful machine learning approaches to arrive at more mechanistically relevant and well-informed in vitro hazard decisions with accuracies superior to animal models.

 

CV Ivan Kopljar

Ivan Kopljar obtained his PhD in Biochemistry from the University of Antwerp, where he studied the molecular pharmacology of voltage-gated potassium channels. In 2013, he joined the Global Safety Pharmacology group at Janssen Pharmaceutica (Johnson & Johnson Pharmaceutical R&D) in Beerse (Belgium) as an IMI STEMBANCC post-doc. His research focused on the application of human stem cell-derived cardiomyocytes (hiPSC-CMs) in preclinical drug safety. He was involved in the development and implementation of hiPSC-CMs-based high throughput screening assays to evaluate acute and chronic cardiac liabilities. Since 2018, he holds a position as study director in the In Vivo Sciences group within the Non-Clinical Safety department at Janssen Pharmaceutica.

 

CV Peter Racz

Dr Racz obtained his PhD at the University of Leiden. He has 15 years of research experience in different fields including biomarker discovery, preclinical studies, and more than five years of experience in Developmental and Reproductive Toxicology (DART) in vitro assay development. He has worked in academia as well as in industry in several countries (USA Netherlands, Hungary). Before his current job at Toxys, he worked as a senior researcher at ZF-screens, a Dutch biotech company specialised in zebrafish research. There, he was leading NC3R Crack it challenge within the company in which he investigated the transcriptome response of the zebrafish larvae upon teratogen compound exposure. In his current job at Toxys, he is developing a novel human stem cell-based gene expression assay for in vitro DART Assessment.

 

CV Andy Forreryd

Andy Forreryd holds a Master of Science in molecular biotechnology from the Faculty of Engineering LTH at Lund University and a Doctor of philosophy in Immunotechnology from Lund University. He has many years of experience working with in vitro assay development, genomics and machine learning. Andy is currently employed by SenzaGen as a scientist. He is co-developer of the GARD platform for assessment of chemical sensitizers and inventor of two patents related to the technology.