Development of new cell lines as a means of reducing animal testing in toxicology

The project overseen by Filipič is aimed at obtaining more accurate controlled tests in a laboratory while reducing the need to use animals as subjects. The NIB, in association with the Ljubljana Oncology Institute and Austrian scientists from the Vienna University of Medicine, are working to develop new cell lines for human liver tissue which could be used in research on the effects of genotoxic factors on the human body.

"If the results aren't realistic, we obtain false positives, which in turn means we have to conduct tests on animals (in vivo tests) unnecessarily. This is why obtaining more accurate tests is so important," Filipič said.

Based on the ideas of Dr Filipič the researchers working on the project have developed an alternative in vitro testing system for quick detection of genotoxicity intended for screening of new compounds in pharmacy, cosmetics and food as well as in environmental monitoring. The method obtained a patent from the Office for Intellectual Property of the Republic of Slovenia in 2011.

The effects of medicine on the environment, organisms and human health has been a subject of extensive research. A key focus of studies is cytostatic medication which is used in chemotherapy for cancer. "Cancerous cells multiply quickly, which is why they are more sensitive to cytostatics. However, these do not work selectively and may also impact healthy cells," said Filipič.

Cytostatics can also adversely affect healthy organisms indirectly when they are released into the environment. After being metabolised, the drug is excreted in feces and urine, from where it enters waterways. The presence of cytostatics and their impact on water organisms was a subject of a four-year study conducted within the EU's Seventh Framework Programme for Research entitled Fate and effects of cytostatic pharmaceuticals in the environment and the identification of biomarkers for and improved risk assessment on environmental exposure - CYTOTHREAT. The EUR 3.5m project was coordinated by Filipič.

One of the important findings of the CYTOTHREAT study, which was concluded in 2014, is that even small quantities of cytostatics in the environment pose a risk of damage to genetic material, which can in turn result in the development of cancer. The project also helped develop new analytical methods which can be used in measuring toxicity and risks associated with cytostatics in the environment.

In Filipič's opinion the main challenge facing toxicology researchers is difficulty in securing projects. "Toxicology requires many basic studies, which should be funded by society. But everything today is focused on industry, which has no interest in fundamental toxicological research because of its focus on delivering products to the market in the shortest possible period."

The Department for Genetic Toxicology and Biology of Cancer has for over 15 years been studying the damaging effects of cyanotoxins, which are produced by cyanobacteria. These are released into water during algae blooms and represent a significant danger to animals and people with their ability to cause DNA mutations and trigger the development of tumors. "It is expected that climate change will increase the presence of toxic cyanobacteria in surface water. For this reason it is important to better understand the danger they present to human health," said Dr Bojana Žegura, who studies cyanotoxins.

Certificate of Good Laboratory Practice

The Department for Genetic Toxicology and Biology of Cancer, which currently employs seven researchers, runs a laboratory which was certified in 2015 for Good Laboratory Practice in line with OECD standards for its work in the study of mutagens. It is the only public laboratory to perform its laboratory tests in line with standards prescribed by the OECD for Good Laboratory Practice, said Filipič. The only other laboratories in Slovenia with this certificate are run by pharmaceutical companies Krka and Lek.

"This is important because genotoxicity tests are mandatory in pharmacy, cosmetics and similar industries. There is a growing need for testing, which is why we decided to establish a laboratory at NIB based on our extensive experience with testing." Slovenian industry has shown a lot of interest, but the institute's plans extend beyond the Slovenian market, Filipič said.

The fundamental premise of the principle of Good Laboratory Practice is traceability of the entire testing process, which ensures high quality and reliable data on human and environmental safety. It also promotes mutual recognition of data obtained in studies among the member countries in order to reduce testing on animals and overall costs.

About Dr Metka Filipič

Dr Metka Filipič has established herself as a prominent researcher in genetic toxicology in recent years. She has published more than 70 original articles in journals with a high impact factor, including in Biosensors & Bioelectronics, Water Research, Journal of Hazardous Materials and Nanotechnology. Filipič is a tenured professor of toxicological chemistry who lectures at several faculties of the University of Ljubljana, including pharmacy and the Jožef Stefan International Post-Graduate School. She has been a mentor to nine doctoral graduates.

Filipič is a member of several editorial boards, including of the journals Reviews in Mutation Research and Food and Chemical Toxicology. She is an active member of numerous domestic and international associations. Since 2002 she has headed the Slovenian government's scientific committee on GMOs and served as the expert member of the programming committee for the Seventh Framework Programme for six years. Between 2009 and 2012 Filipič was a member of the scientific committee on food chain contaminates at the European Food Safety Agency, before becoming a a member of the scientific committee for plant safety in the same agency in 2012.

Cooperation chart for Metka Filipič

Source: Atlas of Science