Non-animal method with regulatory approval for evaluating hormone potency awarded 3Rs Prize

This year’s 3Rs Prize has been awarded to Dr Francesco Nevelli at Merck for a publication describing the development of an in vitro assay for testing follicle-stimulating hormone product potency. The in vitro method was approved by regulators for use in potency testing, at best replacing tens of thousands of animals annually.

Dr Lorna Ewart, Emulate Inc was highly commended for a paper analysing the predictability of an organ-on-a-chip device to determine liver toxicity. In the largest characterisation study of its kind, the paper showed the device had increased sensitivity in identifying toxic therapeutics compared to both current in vitro methods and in vivo studies.

“The applications for this year's 3Rs Prize were truly outstanding and the Panel had a challenging job to do in selecting the winner. The pioneering 3Rs research taking place in academia and industry across the globe is astonishing. This year marks a first with both the winner and the highly commended applications coming from industry. I look forward now to seeing the replacement potential of these new methods being realised in the coming years.”

Professor Julia Buckingham
Chair of the NC3Rs Board and 3Rs Prize Panel

“GSK continues to support the NC3Rs 3Rs International Prize as part of our longstanding commitment to furthering innovation in non-animal methods and technologies, reducing animal use where possible and improving the welfare of laboratory animals. This is the 19th year that GSK has supported the prize and the quality and impact of the winning entries continues to be outstanding”. 

Dr Gina Wilkerson
Chief Veterinary Officer, GSK

Winning paper: A human cell-based assay for fertility hormone testing

Follicle stimulating hormone (FSH) is produced for fertility treatments, most predominantly for IVF to increase the number of eggs available to collect and fertilise. Over 70,000 IVF cycles were reported by the Human Fertilisation and Embryology Authority in the UK alone in 2021. FSH potency can be influenced by its purity and chemical and physical changes introduced during the manufacturing process and as a result each batch of FSH must be screened for its potency as part of regulatory requirements. Until recently this testing was conducted in the Steelman-Pohley bioassay, set out in the European and US Pharmacopoeias, (the collections of standards and quality control assays used by regulatory bodies). In the assay, female rats are injected daily with FSH and the ovaries are subsequently removed and weighed, with ovarian weight increase being used as a measure of potency.

The in vitro assay developed by Francesco and colleagues at Merck can replace the Steelman-Pohley bioassay and in 2022 was approved for use by the European Medicines Agency for two products used in fertility treatments. The assay consists of a human cell line, HEK-293, transfected to express the FSH receptor. Potency is assessed by measuring increased cAMP levels following FSH receptor activation. Comparative studies benchmarking the in vitro assay against the rat model demonstrated a very high level of concordance in a range of parameters essential to assessing potency, including distinguishing chemical and structural changes that affect FSH activity.

Regulatory bodies, including the European Medicines Agency, have approved the use of the in vitro assay in more than 70 countries. Francesco and colleagues are now working towards worldwide acceptance to enable Merck to fully replace all rats used for potency tests of Merck Healthcare marketed products, with estimated saving of nearly 40,000 animals annually, based on forecast animal-use in 2023. The Prize money will be used to fund educational programmes for physicians and patients in the USA to increase acceptance of in vitro quality control processes for biological products.

Highly commended paper: Liver-on-a-chip toxicology studies

Drug-induced liver injury is a common reason why potential therapeutics fail in clinical trials despite preclinical screening in both in vitro and in vivo models. In this year’s highly commended paper, Lorna and colleagues describe a screening strategy with “Liver-Chips”, which helps to avoid the use of animals by identifying toxicities early in the drug development pipeline and therefore preventing compounds destined to fail in the clinic being tested in vivo. The Liver-Chips contain four human cell types with hepatocytes embedded in extracellular matrix in one channel and immune and vascular cells in a separate channel. Culturing the cells in the chips replicates key histological features of the liver, including cuboidal liver cell morphology and bile canaliculi-like structures, and stable expression of liver-specific genes.

To demonstrate the utility of the screening strategy, Lorna and colleagues analysed across a blinded set of 27 known hepatotoxic and non-toxic small molecule drugs the performance of the Liver-Chips, in the largest characterisation study of this kind to date. Liver-Chip screens had no false positives, and successfully identified over 80% of drugs known to cause drug-induced liver injury, outperforming the data obtained from animal studies. This study helps to build confidence in the utility of organ-on-chip technologies in the short-term to help minimise the use of animals in drug testing and in the longer term to replace in vivo testing with complex in vitro approaches.

About the 3Rs Prize

The 3Rs Prize is awarded annually by the NC3Rs for a paper that describes outstanding and original work that has or could have major impacts on the 3Rs. The winning researcher is awarded a £28k grant and a £2k personal award. The highly commended winner receives a £4k grant and a £1k personal award. A £20k contribution towards the winning grant is provided by GSK with all remaining funds, including the personal award made by the NC3Rs. 

The competition is open to any research team in the world and applications are assessed by an expert Panel.

About GSK 

GSK is a global biopharma company with a purpose to unite science, technology, and talent to get ahead of disease together. Further information can be found on GSK’s website. 

About the NC3Rs 

The NC3Rs is a UK-based scientific organisation dedicated to helping the research community worldwide to replace, refine and reduce the use of animals in research. Primarily funded by the UK Government, the NC3Rs is also supported by the charitable and private sectors. It collaborates with organisations from across the life sciences sector, nationally and internationally, including universities, industries, other research funders and regulatory authorities.