Sleepless in Seattle: An update from the World Congress on Alternatives and Animal Use in the Life Sciences

Our experts will be working round-the-clock in Seattle this week at the World Congress on Alternatives and Animal Use in the Life Sciences! Over the next several days, staff from The Humane Society of the United States, Humane Society International & The Humane Society Legislative Fund will be at the vanguard of the discussion at this global event that brings together the world’s foremost experts on animal testing alternatives & animal welfare from industry, government, non-profits & academia.

Are you in Seattle, too? Please visit us at our booth (#309) and plan to attend the following events! Somewhere else in the world? Check out what we’re up to below and view the World Congress program.

SUNDAY, AUGUST 20

• 10:00 a.m. – noon
“Introduction to Adverse Outcome Pathways and the AOP Wiki” Satellite Meeting
Sheraton Seattle, Aspen Room
10:00 a.m. Introduction to the OECD AOP Programme and Online Training course – Kate Willett, Human Toxicology Project Council
10:40 a.m. Building AOPs for Neurotoxicity: Perspective from an Academic – Ellen Fritsche, IUF – Leibniz Research Institute for Environmental Medicine
11:20 a.m. Demonstration and Hands-On Activity with AOP Wiki – Kristie Sullivan, Physicians Committee for Responsibility MedicineSponsors: Human Toxicology Project Consortium and the Physicians Committee for Responsible Medicine

MONDAY, AUGUST 21

• 12:15 p.m. – 1:15 p.m.
“Tracking the successful implementation of Tox21 principles.”
Room 6ABC, Board number A43
Poster presenter Vicki Katrinak looks at how the chemical safety testing landscape has changed over the past ten years, as toxicologists embrace new, non-animal testing strategies.

• 12:15 p.m. – 1:15 p.m.
“Legislative process toward animal testing bans for cosmetics in Brazil.”
Room 6ABC, Board number A44
Poster presenter Antoniana Ottoni discusses how the Brazilian authorities aim to implement validated alternative, non-animal tests for commonly used toxicological endpoints and reviews plans to ensure the regulations are updated before the deadline of 2019.

• 2:00 p.m.
“Implications of the Recent 2016 Amendment of the Toxic Substances Control Act (TSCA) on the Development and Implementation of Non-Animal Methods.”
Session II-2: Government Driven Legislation – EDSP/TSCA; 1:15 p.m. – 2:45 p.m.; Rooms 613-614
Speaker Catherine Willett will explain how animal testing could actually increase under the new chemical safety act, and describe how co-ordinated efforts are needed to exploit non-vertebrate models and other evaluation tools to prevent this.

TUESDAY, AUGUST 22

• 10:30 a.m.
“Challenges in implementing the Frank R. Lautenberg Chemical Safety for the 21st Century Act: A perspective covering stakeholders, the U.S. Congress and the current Administration.”
Session IX-4: Global Regulatory Updates; 10:00 a.m. – noon; Room 607
Speaker: Sara Amundson

• 12:15 p.m. – 1:15 p.m.
“Modeling the human airways: from physiology to pathology.”
Room 6ABC, Board number A65
Poster presenter Lindsay Marshall shows how laboratory models using human cells can be used to look at the function of healthy airways and demonstrates that the models can be adapted to enable the study of airways disease, such as cystic fibrosis.

• 12:15 p.m. – 1:15 p.m.
“iPSC and 3D tissue technologies powerful alternatives to animal models for brain disease research”
Room 6ABC, Board number A77
Poster presenter Marcia Triunfol reports on an exciting, HSI-sponsored workshop held in Brazil earlier this year. Brazilian and international scientists discussed the potential for 21st century technologies to replace animals in brain research.

WEDNESDAY, AUGUST 23

• 12:15 p.m. – 1:00 p.m.
“A ‘bottom-up’ approach to accelerate new opportunities in alternatives and Adverse Outcome Pathway (AOP) practice in China”
Room 6ABC, Board number C81
Poster presenter Tina Qu reports on the progress made in promoting non-animal technologies in China, including the recent roll-out of an AOP-training program.

• 10:45 a.m.
“Towards a 21st-century roadmap for biomedical research and drug discovery”
Session IX-8: Global Efforts Moving Towards Replacement of Animals;
10:00 a.m. – noon; Rooms 619-620
Speaker Troy Seidle describes the application of pathways-based approaches to understand chemical safety and details how a series of workshops sponsored by The Humane Society of the United States and Humane Society International have introduced this concept to other stakeholders – describing the main recommendations from these events.

THURSDAY, AUGUST 24

• 10:48 a.m.
“The Human Toxicology Project Consortium: a private-public partnership to promote development and acceptance of pathway-based science”
Session IX-6: 3Rs Communication and Advocacy; 10:00 a.m. – noon; Rooms 619-620
Speaker Catherine Willett will describe the work of the Human Toxicology Project Consortium (HTPC), which aims to advance a biological pathway-based approach to toxicology. She will explain how HTPC intends to achieve this through scientific activities, communication, and advocating for financial and legislative support.

• 11:45 a.m.
“Deletion of scientifically redundant animal test requirements in the agrochemical sector: the case of the 1-year dog study”
Session IV-1: Predictive Safety Approaches Role in the Design of Inherently Safer Chemicals; 10:00 a.m. – noon; Rooms 615-616
Speaker Marco Corvaro will explain how the ongoing global effort to end a test that has been proven redundant illustrates how challenging it can be to enact regulatory change. He will also discuss strategies to improve the use of modern approaches to testing and risk assessment.

• 11:46 a.m.
“A campaign to end invasive chimpanzee research and retire all chimpanzees to sanctuaries: successful strategies and lessons learned”
Session IX-6: 3Rs Communication and Advocacy; 10:00 a.m. – noon; Rooms 619-620
Speaker Kathleen Conlee will take us from 2006, when The Humane Society of the United States launched their campaign to end the use of chimpanzees in invasive research, to today where numbers of chimps in labs have dropped and research chimps are finally being retired to sanctuaries.

AOP tutorials AOPs non-animal tests toxicity testing alternatives toxicology World Congress on Alternatives
Picture of video reel, by Coyau / Wikimedia Commons / CC-BY-SA-3.0

Animal-free skin allergy testing

With the recent approval of the human Cell Line Activation Test (h-CLAT) for skin sensitization (allergy), toxicologists now have a battery of methods that allows them to test for sensitization without using animals.mice

Testing a chemical substance for skin irritation or corrosion is pretty straight-forward: the substance is applied to a skin sample (there are many non-animal in vitro options) and damage will be seen relatively quickly. But to learn if the chemical has the potential to cause skin allergies, testing is more complicated. Skin sensitization is a two-stage process. In the first stage, a chemical exposure “primes” the immune system. Additional exposures then provoke an allergic response (inflammation, redness, itching, etc). Because of the biological complexity of the process, skin sensitization testing is usually conducted on mice or guinea pigs.

But mice and guinea pigs don’t always react the same way as humans would to potential skin allergens. To replace these animal tests with more human-relevant methods, toxicologists have long recommended developing a battery of in vitro tests that could be combined in an Integrated Testing Strategy – where each test method captures a different part of the skin sensitization process. The biological processes that underlie the skin allergy reaction are pretty well understood, and have been described (the Organisation on Economic Co-operation and Development (OECD) has published a description of this process – the Adverse Outcome Pathway leading to skin allergies). Two of the pieces of this strategy are already in place: the OECD approved the use of the Direct Peptide Reactivity Assay (DPRA) and the KeratinoSens test – each measuring a different step in the sensitization process. The DRPA assays measures whether a chemical can react with proteins in a way that causes the protein to become an allergen. The KeratinoSens assay measures activation of genes involved in the allergic reaction in skin cells (keratinocytes). The h-CLAT method completes the battery of tests: it detects biomarkers that indicate activation of immune (dendritic) cells in the skin.

Many countries require that chemicals used in manufacturing, agriculture, medicine, and cosmetics be tested for their potential to cause skin allergies in humans. Without approved in vitro options, REACH regulations in the EU alone would force industry to use hundreds of thousands of animals for skin sensitization testing. The animal-free, 3-test strategy is a “textbook” example of the mechanistic, pathway-based approach to chemical testing promoted by the Human Toxicology Project Consortium. Use of this strategy has the potential to greatly reduce the numbers of animals used for skin sensitization testing, while also reducing the cost and time it takes to produce human-relevant results.

alternative toxicity testing h-clat skin sensitization toxicity testing alternatives

“Blood-brain barrier on-a-chip”

Scientists at the Wyss Institute have created a 3-dimensional in vitro model of the human blood-brain-barrier (BBB)-“on-a-chip.”  The device will make it possible for researchers to test drugs, chemicals, and disease factors that interact with the BBB, without using animals – and in a 3-dimensional environment that mimics that of the human BBB in vivo.

The BBB is a semi-permeable cellular structure that allows some nutrients and substances to enter the blood flow in the brain, and keeps other elements (such as bacteria and potential toxins) out. Because it is so effective, it can also prevent useful treatments from reaching targets in the brain. Researchers need to understand how and why certain substances can pass through the barrier, in part so they can design therapeutic drugs accordingly, and so they can design other substances to prevent neurotoxicity.

From the Wyss Institute press release: "These fluorescence confocal microscopy images show both a high magnification view (left) of a region of the human brain capillary endothelium within the endothelium lined tube (shown at lower magnification at right) that, in combination with surrounding human pericytes and astrocytes, comprise the blood-brain barrier (cell junctions linking adjacent endothelial cells are shown in magenta). " Credit: Wyss Institute at Harvard University

From the Wyss Institute press release: “These fluorescence confocal microscopy images show both a high magnification view (left) of a region of the human brain capillary endothelium within the endothelium lined tube (shown at lower magnification at right) that, in combination with surrounding human pericytes and astrocytes, comprise the blood-brain barrier (cell junctions linking adjacent endothelial cells are shown in magenta). ” Credit: Wyss Institute at Harvard University

To create the device, the Wyss Institute team carved a tiny channel in a polymer chip and filled it with a gel matrix containing human astrocytes, the cells that comprise the extra-tight “barrier” around blood vessels in the brain. Another channel was tunneled through this matrix and seeded with human pericyte cells (contractile cells which control the “gaps” through which substances can enter the neurological bloodstream) and then with human endothelial cells (the cells that line the interior of a blood vessel). The cells “self-assembled” into the same layers and connections they exhibit in blood vessels in vivo.

To test the model, the team introduced a protein known to cause inflammation – one that has been associated with a number of central nervous system diseases and disorders, including Alzheimer’s, multiple sclerosis, and stroke (among others). The in vitro BBB responded by producing protective proteins. The device can thus be used to study neuroinflammation, and to test new treatments.

Read more in the Wyss Institute’s press release.

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Educational Infographic produced by the Human Toxicology Project Consortium

THE FUTURE OF TOXICITY TESTINGHTP_infographic_FINAL_revised

A new infographic produced by the Human Toxicology Project Consortium shows in three sections how the future of toxicity testing promises a steady reduction in testing costs, increases in human relevance and confidence in safety assessments, and the eventual elimination of animal tests.

The first section provides a snapshot comparison of the current and future costs, efficiency and efficacy of toxicity testing, while the mid portion uses pesticide testing as a specific example of now, vs near-future, vs the optimal approach that, given the focus and resources necessary, will be envisioned within the decade.

The near-future and optimal approaches rely increasingly on our understanding of biology and using it to build a predictive systems biology platform that is comprised of an interrelated network of biological pathways. This platform is used to design and interpret tests that provide much more efficient and effective characterization of chemical activity that can be used to predict safe use of chemicals.

Finally, the results of this progression are captured in the summary graphic at the end – decreasing costs, animal use and time while human relevance and our confidence in safety decisions continue to improve.

As explained on our Project page, the Human Toxicology Project Consortium works on three areas critical for the successful, international implementation of a pathways-based approach to chemical safety testing: advancing the science, communicating the purpose and goals of pathway-based toxicology, and lobbying for funding and policy changes that will support pathway-based approaches in the US and around the world.

To advance our communication and education efforts, HTPC member organizations worked together to create this infographic, to quickly and effectively illustrate the differences between traditional animal-based toxicity testing and pathway-based testing in terms of predictive power, cost, and testing capacity.

Details on the numbers used in this comparison are available here (PDF).

alternative toxicity testing AOPs computational toxicology databases drug discovery EPA HTPC members in the news non-animal tests non-predictive animal models pathway-based approaches regulatory toxicology ToxCast toxicity testing alternatives

Chipping away at the use of animals to predict human diseases

The Wyss Institute recently announced two new human cell-based inflammatory disease models built on its rapidly expanding “organ chip” platform. Both models could speed the development of treatments for these diseases, and further reduce the use of animals in testing.

Using the “gut-on-a-chip” device first introduced in 2012, Wyss scientists co-cultured human intestinal cells with normal and pathogenic intestinal microbes, producing an in vitro model (viable for up to two weeks) of intestinal inflammation and bacterial overgrowth. These two disease features are present in a number of human intestinal disorders (such as ulcerative colitis and Crohn’s disease). Until now, it has been difficult to reproduce these disorders in the lab in order to test treatments for them. The gut-on-a-chip device “could allow breakthrough insights into how the microbial communities that flourish inside our GI tracts contribute to human health and disease.” The image below, from the Wyss Institute press release, shows how the cells in this microenvironment even reproduce normal peristalsis, the contraction/relaxation cycle of the intestinal walls that moves digested food down the tract.
gut-stretching-400x298

The Wyss Institute also used its chip technology to create a human lung “small-airway-on-a-chip.” When the chips are lined with airway cells from patients suffering from such inflammatory disorders as chronic obstructive pulmonary disease (COPD) or asthma, the physiological features of the disease can be observed and tested in vitro. As noted in the Wyss Institute press release, “Demand for such opportunities is especially high since small airway inflammation cannot be adequately studied in human patients or animal models and, to date, there are no effective therapies that can stop or reverse the complex and widespread inflammation-driven processes.”

Watch a video demo of the small-airway-on-a-chip below:

3D cell & tissue culture organs-on-chips toxicity testing alternatives