Project: A novel in vitro Screening system for the prediction of Drug Induced Liver Injury

Drug-induced hepatotoxicity is the leading cause of acute and chronic liver failure and the most common adverse event causing drug non-approval and withdrawal by the European Medicine Agency (EMEA) and the U.S. Food and Drug Administration (FDA). Approximately 40% of new drug candidates fail in clinical trials because of serious toxic events that reCOed unrecognized in preclinical studies. _x000D__x000D_The current gold standard for the prediction of human hepatotoxicity is still animal studies that are largely hampered by high cost, bad predictive value and a common ethical problem. While 85% of cardiovascular, 88% of gastrointestinal and 90% of hematological toxicity can be predicted by animal toxicity tests, hepatotoxicity prediction still suffers from less than a 50% accuracy and is therefore recognized as inappropriate for drug screening campaigns. _x000D__x000D_There is clear need for reliable in vitro tools with good predictive power, human relevance, fast turnaround times, and broader, more rapid accessibility. Goal of the present project is to fill this vacancy by the DILI-Screen, a novel in vitro screening system for the prediction of Drug Induced Liver Injury by using cryopreserved human hepatocytes (CryoHeps), single or pooled, and multiple toxicity endpoint measurements. We attempt to develop a first working prototype of the DILI-Screen within this project. _x000D__x000D_Human hepatocytes have largely been recognized as the most relevant in vitro test system for predicting human hepatotoxicity and quickly became the industry’s prime candidate for a new gold standard. Despite the broadly recognized value of this system, one major obstacle reCOs the scarce and unpredictable access to human livers and the fact that the hepatocytes isolated from the livers can COtain full hepatic activity only for a short period of time and therefore need to be used within few hours of preparation. This has prompted researchers to search for alternative cellular models that have greater availability, better storage performance and better reproducibility. Permanent human hepatocyte cell lines such as HepG2 or HepaRG are currently discussed as alternatives. Although there is emerging evidence that these cells have potential for preclinical screening, their use reCOs limited primarily because they are immortalized cells with dramatically different biological features compared to hepatocytes isolated from human liver. Furthermore, these cells represent only a single phenotype and therefore cannot mirror the diverse spectrum of hepatic activity in patients._x000D__x000D_The aim of the present project is to develop, validate, and market DILI-Screen, a screening tool that uses cryopreserved human hepatocytes (CryoHeps), from single donors or pooled. The long-term objective is to make the tool available in two variations: DILI-ScreenAT will address acute toxicity while DILI-ScreenCT will address chronic toxicity. Both systems will be based on CryoHeps with high post-thawing viability and broad hepatic activity. The two participating EUROSTARS SME, KaLy Cell (KLC, Alsace- France) and Pharmacelsus (PHC, Saarland - Germany) will bundle their expertise in human hepatocyte culture and preclinical testing and generate a transnational resource for the pharmaceutical industry that can be accessed from around the world. The two SME are complementary in the sense that KLC has developed a strong expertise on human hepatocytes and their use in the field of ADMET-TOX, while PHC has developed highly sensitive analytical tools for assessment of cellular functions and industry-standard ADME-Tox services under GLP-standard. During the present project, the consortium will first develop and market the DILI-ScreenAT version of the kit. Based on the results obtained in DILI-ScreenAT project, we will further develop the follow-on kit DILI-ScreenCT , which will require plateable CryoHeps produced by KLC. _x000D__x000D__x000D_

Acronym	DILI-Screen (Reference Number: 5474)
Duration	01/09/2010 - 31/07/2013
Project Topic	The aim of the project carried out by a French and a German SME is to develop, validate, and market DILI-ScreenAT, a novel screening tool that uses cryopreserved human hepatocytes (CryoHeps) from single donors or pooled to predict drug induced liver injury in vitro.
Project Results (after finalisation)	One of the most important characteristics of the final test kit was reproducibility. It was possible to show an excellent reproducibility by the results for the tested compounds obtained at PHC and KLC. Moreover, it was possible to obtain reliable data with different batches of CryoHeps. All data (10 reference compounds, final SOPs, batch P2209A) have been analysed.
Network	Eurostars
Call	Eurostars Cut-Off 4

Project partner

Number	Name	Role	Country
2	KaLy Cell	Coordinator	France
2	Pharmacelsus GmbH	Partner	Germany