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The GARD assay for skin and respiratory sensitisation

Andy Forreryd, Lund University
Aug 05, 2015 at 11:00 AM EDT.

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In order to reduce exposure to sensitizing compounds, it is of high importance to identify possible chemical allergens before they reach the consumer market or working environments. Therefore, a significant effort is currently invested in the development of predictive assays for proactive identification of such compounds.

We have developed a cell based testing strategy for assessment of both skin and respiratory sensitizers. The assay was initially designed for the risk assessment of skin sensitizing compounds. A panel of reference compounds comprising skin sensitizing chemicals (N=20) and controls (N=20) were used for stimulation of cells. Complete transcriptomes of cells were analyzed using microarrays, and a genomic biomarker signature was identified using one-way ANOVA p-value filtering, and further optimized using an in-house developed wrapper algorithm. The applicability domain of GARD was later broadened to cover prediction of respiratory sensitizers, using a panel of reference compounds comprising respiratory sensitizers (n=10) and controls (n=22), with a similar methodology as described for skin sensitizers.

Performances of the biomarker signatures were validated using external test sets. The biomarker signatures were challenged with either 37 chemicals (skin sensitization), or 25 chemicals (respiratory sensitization). Accuracy, sensitivity and specificity were estimated to 89%, 92% and 83% for skin sensitization, and 84%, 67% and 89% for respiratory sensitization, respectively.

The transcriptome-wide profiling used during the construction of the assay also provides the possibility to further broaden the applicability domain of GARD to cover also additional endpoints or relative potency. Analyzing the complete transcriptome of cells also provides the benefit of being able to perform mechanistic studies in order to elucidate molecular pathways that drives the pathogenesis of allergic diseases.

Integrated Approaches to Testing and Assessment: Promises

and Challenges of a More Flexible Approach to Toxicology Testing
4th Annual ASCCT Meeting

October 1-2, 2015
Environmental Protection Agency
Durham, NC

Plan to join your colleagues for the next ASCCT annual meeting, which will be held at the Environmental Protection Agency in the Research Triangle Park area of North Carolina.

Abstract submission deadline: July 15

Day 1

• Moving Beyond One Test: Leveraging the Whole Toolbox for Integrated Decision Strategies, Warren Casey, NICEATM
• Invited IATA application case studies

  1. Developing non-testing IATA informed by mechanistic insights: Case studies in building scientific confidence, Grace Patlewicz, Environmental Protection Agency
  2. OECD Implementation of IATA Demonstrated by the Skin Sensitization Endpoint, Johanna Matheson, Consumer Product Safety Commission
  3. Why and How Do We Incorporate Metabolism and Kinetics in Integrated Testing and Assessment, Miyoung Yoon, The Hamner Institutes for Health Sciences

• Poster viewing + flash oral presentations
• Oral presentations from submitted abstracts
• ASCCT Business meeting + Social reception

Day 2

• Chemicals and Risk: New Approaches to Current Practices, Craig Rowlands, Dow Chemical Company
• Additional oral presentations from submitted abstracts
• Discussion panel on IATA topics

Registration and abstract submission details can be found here.

FutureTox III: Transforming 21st Century Science into Risk Assessment and Regulatory Decision-Making
November 19–20, 2015
Arlington, VA

Past Educational Member Webinars

ASCCT members, click here to access recordings of past webinars listed below. Contact the Secretary for more information or to suggest a topic for a future webinar.

Past Events

Second Annual Scientific Meeting of the American Society for Cellular and Computational Toxicology: The Future Is Here: Practical Applications of Emerging Scientific Tools
Human Organs on Chips as Replacements for Animal Testing, the keynote address given by Donald E. Ingber, M.D., Ph.D., of the Wyss Institute at Harvard, kicked off the meeting which featured a dozen lectures and poster presentations on making toxicology a more human-relevant science. Other topics included methods to replace live animals in eye irritation tests and computer-based virtual embryos that show how chemical exposures might affect developing embryos. View Program. ASCCT members, click here to view select presentations.

First Annual Scientific and Business Meeting of the ASCCT
The first annual ASCCT meeting focused on advances in the fields of in vitro and computational toxicology. Dr. Melvin Andersen, associate director of The Institute for Chemical Safety Sciences at The Hamner Institutes for Health Sciences and ASCCT board member, gave a plenary lecture on Computational Cellular Pathway Modeling: Combining Key In Vitro and In Silico Tools to Enhance Modern Safety Assessment. Dr. Suzanne Fitzpatrick, senior advisor for toxicology at the U.S. Food and Drug Administration gave a plenary lecture on an Overview of the FDA-DARPA-NIH Collaboration on Human/Organ on a Chip.

See attached program for abstracts and agenda. A podcast of the event will be coming soon. For a recording of the webinar on the topics discussed at the in-person business meeting, click here.

Institute for In Vitro Sciences Practical Methods for In Vitro Toxicology Workshop
Attendees gained hands-on laboratory experience and instruction in the practical application of the lectured topics. Topics for the three and a half day course included: (i) international regulatory acceptance status of in vitro assays, (ii) predicting organ specific toxicity (e.g., ocular, dermal and hepatic), (iii) using cell and organ culture models with histological, biochemical, and molecular endpointsm, and (iv) data interpretation.

2010 In Vitro Alternatives Forum
Thank you for contributing to a successful forum. Read the post-meeting report.

8th World Congress for Alternatives and Animal Use
Thank you for helping to make the ASCCT/ESTIV luncheon session a success. A manuscript outlining the discussions that took place during the session has been published. You can view the entire proceedings here.