Our Alternatives to Animal Testing (ATAT) programme began in 2016. Over the last half-decade, we’ve been exploring the potential of this approach to assess next generation product (NGP) aerosols compared to cigarette smoke.
But how has the shift away from using animals in science come to pass?
Animal testing has been prevalent in biological research since the 1950s, and many scientists still rely on these relevant, but arguably unethical, techniques. The approach started to change around two decades ago, when the US National Research Council (NRC) began to recognise rapid advances in biotechnology.
In 2007, the NRC outlined a strategic plan to advance toxicity testing through new methodologies that evaluated changes in biological processes using cells, cell lines, or cellular components of human origin – rather than traditional in-vivo methods using live animals.
They called it Toxicity Testing in the 21st Century, or TT21C.
Since then, those manufacturers and scientists who support the movement to use fewer animals in research in laboratories across the world have made seismic strides in proposing and developing cutting-edge alternative methods that are more human-relevant.
That includes our own industry – and we’re proud to be among those pioneering a TT21C-driven approach, aligning with our wider corporate position not to test on animals unless required by regulatory authorities.
Our own TT21C research programme – ‘Alternatives to Animal Testing’ (ATAT) – began in 2016. During the last half a decade, we’ve been exploring the potential of this approach to assess next generation product (NGP) aerosols compared to cigarette smoke.
The results have been very promising, highlighted by the acceptance of our studies into the peer-reviewed academic literature.
For instance, in conjunction with recognised regulatory in-vitro test methods, we’ve examined the effect of our NGP on cardio endpoints. Watch the video below to learn more.
We’re also using a range of other assays to assess cancer and chronic respiratory disease endpoints, heart disease, organ interactions, and in-vitro models to populate parts of adverse outcome pathways (AOP).
You can explore them in our interactive table at the bottom of this page.
Using multiple assays related to different areas of smoking-related disease allows a weight-of-evidence approach to help draw compelling conclusions around the harm reduction potential of our NGP.
ATAT has incredible potential, but it’s still an emerging framework in some ways – not least because it’s still less than two decades young. This means there’s still lots to do in terms of educating regulators, media, and the public around its advantages.
However, we’re confident many ATAT methods and assays will become established as future regulatory standards.
Several agencies are demonstrating keenness to encourage this shift: the most high-profile being the U.S. Food and Drug Administration (FDA), who have previously acknowledged the need for New Assessment Methods (or NAMs) through their own Tox21 programme and roadmap.
Meanwhile, People for the Ethical Treatment of Animals (PETA)’s Science Consortium have commended our research programme for its TT21C-driven approach.
There has been astonishing progress in the development of human-relevant non-animal testing approaches, and their use to more reliably and rapidly provide information for predicting the potential toxicity of chemicals to humans. We support the advancement of scientifically sound non-animal testing approaches, as demonstrated by Imperial Brands, and partner with others fostering their advancement.
Andreas Stucki, PhD
PETA Science Consortium International e.V.
https://www.thepsci.eu/
We continue to scientifically engage with regulators to build the knowledge around, and acceptance of, approaches like ATAT to help enable more ethical and human-relevant assessments in the future.
TT21C is a real toxicological gamechanger and our Alternatives to Animal Testing team is proud to work on this cutting-edge harm reduction science.
Matt Stevenson
Scientific Substantiation Senior Manager
Established regulatory assays
Before any advanced techniques are used, our products are assessed in standardised assays and tests endorsed by regulatory bodies across the world. These assess cytotoxicity, genotoxicity and mutagenicity – all of which are important for understanding the potential toxicity of the product and how it compares to cigarettes.
In-vitro Micronucleus (IVM): The IVM assay detects damage to the structure of cellular DNA (genotoxicity) following test article exposure. Genotoxicity has been mechanistically linked with the formation of cancer.
You can read more about this research here.
Established regulatory assays
Before any advanced techniques are used, our products are assessed in standardised assays and tests endorsed by regulatory bodies across the world. These assess cytotoxicity, genotoxicity and mutagenicity – all of which are important for understanding the potential toxicity of the product and how it compares to cigarettes.
Neutral Red Uptake (NRU): This cell-based approach assesses the uptake of red dye to quantify the number of living cells in culture. The amount of cell damage or death (cytotoxicity) is assessed following exposure to increasing concentrations of test article.
Cytotoxicity is considered a potential step in several chronic disease processes associated with smoking, including cancer, cardiovascular disease and emphysema.
We’ve performed numerous pieces of research, some of which is detailed in this presentation, using these established assays. They all indicate NGP have different scientific profiles compared to cigarette smoke, and offer significant harm reduction potential for adult smokers.
Organ-on-a-Chip (OoC): involves creating miniature, living models of human organs on small chips to study their functions and responses to drugs. These chips are designed to mimic the complex environment of real organs, including blood flow and tissue interactions.
We’ve used this technology to replicate key events of atherosclerosis, a condition that leads to heart disease.
Read more here.
This suite of stem-cell based assays provides insight into the mechanisms by which chemicals can interact with cells, using mechanisms that potentially cause DNA damage compared to those such as protein folding that do not interact with DNA. These assays help assess how product samples may impact cellular functioning across various reporter cell lines, picking up the tell-tale molecular signs of potential harm.
Indicators may include oxidative stress, DNA and protein damage, as well as activation of the p53 gene that has a role in cell cycle regulation and tumour suppression.
Our published research using this methodology shows that undiluted vape e-liquids, and their aerosol extracts, exhibited entirely absent or vastly reduced indications of potential DNA damage in cells, compared to smoke from cigarettes.
Our 2021 ToxTracker study was the first to publish results using the assay for the assessment of vape e-liquids and aerosols.
This assay combines automated imaging and quantitative data analysis to provide rich systems biology information, detecting cellular events in human cell lines in response to test materials.
In numerous pieces of research using this methodology, we’ve shown that vape e-liquids elicit lower biological activity when compared to cigarette smoke.
Read more here.
This assay combines automated imaging and quantitative data analysis to provide rich systems biology information, detecting cellular events in human cell lines in response to test materials.
In numerous pieces of research using this methodology, we’ve shown that vape e-liquids elicit lower biological activity when compared to cigarette smoke.
Read more here.
With the use of in-vitro new approach methodologies (NAMs) for the assessment of NGP, new extrapolation methods will also be required to interpret and contextualise the physiological relevance of these results.
Quantitative in-vitro to in-vivo extrapolation (QIVIVE) can translate in-vitro concentrations into in-life exposures with physiologically-based pharmacokinetic (PBPK) modelling, providing estimates of the likelihood of harmful effects from expected exposures.
Read more here.
