Andy Forreryd1, Joshua Schmidt1, Robin Gradin1, Florence Burleson, Helge Gehrke3, Henrik Johansson1. 1SenzaGen AB, Lund, Sweden. 2Burleson Research Technologies, Morrisville, NC, USA. 3Eurofins BioPharma Product Testing Munich GmbH, Planegg, Germany.
Sensitization of the respiratory tract by sensitizing chemicals may lead to severe bronchoconstriction and asthma-like symptoms with a potentially fatal outcome. However, proactive identification and characterization of respiratory sensitization hazards is currently hampered by the lack of validated or even widely used predictive assays.
The GARD™ platform utilizes exposure-induced gene expression profiles of a dendritic cell-like cell line in combination with machine learning as a basis for predictive in vitro assays for different immunotoxicity endpoints (Johansson et al., 2020). Functional evidence of the ability to accurately identify respiratory sensitizers using the GARD platform has previously been demonstrated (Forreryd et al., 2015). Here, we present GARDair, an adaptation of previous findings on a gene expression analysis platform suited for routine acquisition, based on an optimized predictive biomarker signature. Furthermore, we present the initial results of an inter-laboratory ring-trial, demonstrating the functionality, transferability and reproducibility of the assay.
Conclusion
GARDair is a novel in vitro assay for assessment of respiratory sensitizers, based on evaluation of exposure-induced gene expression of genomic biomarkers in a Dendritic Cell-like cell and machine-learning assisted classification.
GARDair is the first in its class to be subjected to an inter-laboratory ring trial.
The method is highly specific, with high PPV, making it suitable for opt-out applications within product development and candidate prioritization.
https://senzagen.com/wp-content/uploads/2020/09/SOT2020-GARDair.png42308554Tina Dackemark Lawessonhttps://senzagen.com/wp-content/uploads/2017/01/LoggaSquare_RGB_liten_HF.pngTina Dackemark Lawesson2020-03-19 15:50:352021-04-23 11:31:38Inter-laboratory ring trial of the GARD™air assay for assessment of respiratory sensitizers
One of Sweden’s most famous influencers, Margaux Dietz, has visited SenzaGen in Lund.
With support from the EU, SenzaGen has developed the world’s first animal-free test for respiratory allergies making our daily products safer. Margaux put a lab coat on and got to allergy test her own shampoo. Now, Margaux knows that the EU investments in innovations make a difference in everyday life.
Abstract Electronic cigarettes (e-cigarettes) are an increasingly popular alternative to combustible tobacco cigarettes among smokers worldwide. A growing body of research indicates that flavours play a critical role in attracting and retaining smokers into the e-cigarette category, directly contributing to declining smoking rates and tobacco harm reduction. The responsible selection and inclusion levels of flavourings in e-liquids must be guided by toxicological principles. Some flavour ingredients, whether natural extracts or synthetic, are known allergens. In this study, we used the Genomic Allergen Rapid Detection (GARD) testing strategy to predict and compare the respiratory and skin sensitising potential of three experimental and two commercial e-liquids. These novel, myeloid cell-based assays use changes in the transcriptional profiles of genomic biomarkers that are collectively relevant for respiratory and skin sensitisation. Our initial results indicate that the GARD assays were able to differentiate and broadly classify e-liquids based on their sensitisation potential, which are defined mixtures. Further studies need to be conducted to assess whether and how these assays could be used for the screening and toxicological assessment of e-liquids to support product development and commercialisation.
https://senzagen.com/wp-content/uploads/2020/07/Sc-Pub.png168172Tina Dackemark Lawessonhttps://senzagen.com/wp-content/uploads/2017/01/LoggaSquare_RGB_liten_HF.pngTina Dackemark Lawesson2019-02-05 14:47:392020-07-09 09:32:25The use of Genomic Allergen Rapid Detection (GARD) assays to predict the respiratory and skin sensitising potential of e-liquids
Henrik Johansson, Andy Forreryd, Robin Gradin, Angelica Johansson, Olivia Larne, Emil Aaltonen, Anders Jerre, Carl A.K. Borrebaeck and Malin Lindstedt
SenzaGen AB, Lund, Sweden. Department of Immunotechnology, Lund, Sweden.
Introduction
Exposure to chemicals may induce allergic hypersensitivity reactions in skin or respiratory tract. To minimize exposure, chemicals are routinely screened for their sensitizing potential. Proactive identification has historically been performed using animal models, but the use of animals for safety assessment of cosmetics was recently banned within EU. Today, similar trends are spreading both globally and across industry and market segments. Methods for specific identification of respiratory sensitizers are greatly underdeveloped, with no validated, or even widely used assay readily available. Thus, there is an urgent need for development of non-animal-based methods for hazard classification of respiratory sensitizing chemicals.
GARD– Genomic Allergen Rapid Detection – is a state of the art technology platform for assessment of chemical sensitizers (Figure 1). It is based on a dendritic cell (DC)-like cell line, thus mimicking the cell type involved in the initiation of the response leading to sensitization. Following test chemical exposure, induced transcriptional changes are measured to study the activation state of the cells. These changes are associated with the immunological decision-making role of DCs in vivo and constitutes of e.g. up-regulation of co-stimulatory molecules, induction of cellular and oxidative stress pathways and an altered phenotype associated with recognition of xenobiotic matter. By using state-of-the-art gene expression technologies, high informational content data is generated, that allows the user to get a holistic view of the cellular response induced by the test substance.
Conclusion
GARDair is a novel assay for assessment of respiratory sensitizers. It is an adaptation of the GARD platform, utilizing gene expression analysis of predictive biomarker signatures and state-of-the-art data analysis methodology. GARDair has been proven functional and is currently progressing towards industrial implementation and regulatory acceptance with financial support from the EU programme Horizon 2020. This progress includes scientific verification of results, assay optimization, assay
transfer and formal validation by a blinded ring trial.
https://senzagen.com/wp-content/uploads/2018/10/GARDair_ESTIV_Poster.jpg8291087admin_senzagenhttps://senzagen.com/wp-content/uploads/2017/01/LoggaSquare_RGB_liten_HF.pngadmin_senzagen2018-10-17 14:17:052021-04-23 15:33:35GARD™air – A novel assay for assessment of chemical respiratory sensitizers based on a genomic platform
a novel in vitro assay based on a genomic biomarker signature
Henrik Johansson, Andy Forreryd, Robin Gradin, Angelica Johansson, Olivia Larne, Emil Aaltonen, Anders Jerre, Carl A.K. Borrebaeck and Malin Lindstedt
SenzaGen AB, Lund, Sweden. Department of Immunotechnology, Lund, Sweden.
Introduction
Exposure to chemicals may induce allergic hypersensitivity reactions in skin or respiratory tract. To minimize exposure, chemicals are routinely screened for their sensitizing potential. Proactive identification has historically been performed using animal models, but the use of animals for safety assessment of cosmetics was recently banned within EU. Today, similar trends are spreading both globally and across industry and market segments. Methods for specific identification of respiratory sensitizers are greatly underdeveloped, with no validated, or even widely used assay readily available. Thus, there is an urgent need for development of non-animal based methods for hazard classification of respiratory sensitizing chemicals.
GARD – Genomic Allergen Rapid Detection – is a state of the art technology platform for assessment of chemical sensitizers (Figure 1). It is based on a dendritic cell (DC)-like cell line, thus mimicking the cell type involved in the initiation of the response leading to sensitization. Following test chemical exposure, induced transcriptional changes are measured to study the activation state of the cells. These changes are associated with the immunological decision-making role of DCs in vivo and constitutes of e.g. up-regulation of co-stimulatory molecules, induction of cellular and oxidative stress pathways and an altered phenotype associated with recognition of xenobiotic matter. By using state-of-the-art gene expression technologies, high informational content data is generated, that allows the user to get a holistic view of the cellular response induced by the test substance.
Conclusion
GARDair is a novel assay for assessment of respiratory sensitizers. It is an adaptation of the GARD platform, utilizing gene expression analysis of predictive biomarker signatures and state-of-the-art data analysis methodology. GARDair has been proven functional and is currently progressing towards industrial implemetation with financial support from the EU programme Horizon 2020. This progress will include scientific verification of results, assay optimization, transfer and formal validation.
https://senzagen.com/wp-content/uploads/2017/01/scientific-publication.png6801200admin_senzagenhttps://senzagen.com/wp-content/uploads/2017/01/LoggaSquare_RGB_liten_HF.pngadmin_senzagen2018-07-20 10:00:102021-04-23 15:37:20Prediction of chemical respiratory sensitizers using GARD™
On July 5th-6th, SenzaGen will be an attending sponsor at LIVe 2018 (Lung In Vitro event for innovative & predictive models) in Nice, France. The meeting intends to be a unique exchange platform for scientists interested in in vitro respiratory researches, stakeholders from academia, pharma, biotech, chemical, tobacco, consumer goods, medical devices industries, CROs and regulatory agencies.
LIVe 2018 focuses on predictive in vitro lung models designed for basic research, efficacy, sensitization and toxicity testing. It aims to present the state-of-art of in vitro lung models, and to map out the route for its future development and use.
Henrik Johansson, PhD, Senior scientist at SenzaGen, will be giving a key note presentation on the abstract “How do we incorporate recent developments in Big Data analysis and Machine Learning technologies in the field of Toxicology?” on July 6th, opening the 6th event session, “Think Different”. This will be followed by a panel discussion, including Henrik Johansson, regarding “Next Horizon for in vitro lung modeling”.
SenzaGen will also be presenting a poster with the title “GARDair – A novel assay for assessment of chemical respiratory sensitizers based on a genomic platform”. The poster summarizes SenzaGen’s findings so far regarding Horizon 2020.
If you will be attending LIVe 2018 as well, please come visit SenzaGen’s booth. Hope to see you there!
https://senzagen.com/wp-content/uploads/2018/07/LIVe_SenzaGen.png595595admin_senzagenhttps://senzagen.com/wp-content/uploads/2017/01/LoggaSquare_RGB_liten_HF.pngadmin_senzagen2018-07-02 09:25:502018-07-16 07:44:18SenzaGen to present at LIVe 2018 in Nice, July 5th-6th
Matthew Stevenson, Lukasz Czekala, Liam Simms, Nicole Tschierske, Henrik Johansson, Tanvir Walele
Imperial Tobacco Ltd, Reemtsma Cigarettenfabriken GmbH, an Imperial Brands PLC Company, SenzaGen AB, Fontem Ventures B.V., an Imperial Brands PLC Company,
Introduction and Objectives
There is a general consensus amongst the scientific and public health community that e-cigarettes constitute a less harmful source of nicotine than combustible cigarettes, and that flavours play a critical role in attracting and retaining smokers into the vaping category. Due to the dynamic nature of innovation with e-cigarettes new assays are required to quickly determine the subtle biological response of these products for product stewardship activities. The size of this task is considerable as recent estimates state that more than 8,000 e-liquid flavours are on the market (Hartung, 2016). One particular toxicological endpoint which is of interest for the Stewardship of e-liquids, is Respiratory Sensitisation.
Respiratory sensitization (RS) is an allergic type I hypersensitivity reaction of the upper and lower respiratory tract caused by an immune response triggered by low molecular weight compounds or other environmental proteins. Clinical symptoms of RS include asthmatic attacks, bronchoconstriction and wheezing upon repeated exposure to the same compound. However, respiratory sensitisers are rare, with around 100 well characterised substances described in the literature.
It is Fontem Ventures policy to screen all novel e-liquid ingredients for Respiratory sensitising activities using published literature and in silico techniques. However, there is a need for alternative techniques to fill data gaps and add to a weight of evidence. Several in vitro assays have been described and validated to assess skin sensitisation, however for respiratory sensitization there are no validated predictive assays. It is of note that not all skin sensitizers are also respiratory sensitizers. In 2015, Basketter and Kimber concluded that “…airborne fragrance materials, including skin sensitising fragrance materials, do not pose a risk of the induction or elicitation of allergic reactions consequent upon exposure via the respiratory tract”. Therefore, it is critical that any assays developed to determine the sensitising properties of a chemical can distinguish between dermal and respiratory activity.
The objective of this study was to assess experimental and commercial e-liquids in GARDair™; an assay which claims to detect respiratory sensitisers.
GARDair measures the genomic biomarker signature of a human myeloid leukemia-derived cell line exposed to test substances; making this technology in keeping with the 3Rs (Reduce, Replace and Refine) and Toxicity Testing in the 21st Century principles. Gene expression analysis is performed using Affymetrix microarray technology and a prediction model is used to classify each sample according to its respiratory sensitizing potential.
Conclusions
• From the Benchmark Control data it was estimated that GARDair™ had a sensitivity and specificity of 71% and 100% respectively; with an overall predictive accuracy estimated as 89%.
• Extensive validation of this assay is ongoing, however, the lack of well characterised Chemical Respiratory Sensitisers may limit this.
• None of the experimental or commercial samples were classified as respiratory sensitisers.
• Further exploration of this assay is required, particularly its ability to detect low concentrations of sensitiser in complex mixtures and to ensure that the e-liquid matrix does not interfere with the detection of activity.
https://senzagen.com/wp-content/uploads/2017/01/scientific-publication.png6801200admin_senzagenhttps://senzagen.com/wp-content/uploads/2017/01/LoggaSquare_RGB_liten_HF.pngadmin_senzagen2018-04-20 10:00:472021-12-03 10:24:49The In Vitro assessment of Respiratory Sensitisation Potential of Electronic Cigarette Liquids
SenzaGen has signed a collaboration agreement with the global biopharmaceutical company AstraZeneca. The collaboration focuses on testing and development of SenzaGen’s sensitization test GARDair, in order to distinguish between allergens and irritants in the respiratory tract. The goal is the validation of in vitro toxicological tests that could be applied to the discovery of new treatments for respiratory diseases without respiratory tract irritancy as a side effect.
The collaboration means that SenzaGen will use its leading technology platform GARD, with the help of AstraZeneca’s well-documented substance libraries, to develop a new genomic signature to distinguish allergens from irritants in the respiratory tract, something which is not possible today with in vitro methods. GARD examines and finds allergens in chemicals and medicines through a unique technology in accordance with the international 3Rs principle, Reduce, Replace and Refine, as an alternative to animal testing.
“During our research and drug discovery activities, it is important to be able to identify and mitigate potential toxicological issues as early as possible, to ensure our novel candidate drugs have the right safety profile for patients. This requires in vitro toxicological methods that are properly validated for their predictivity of clinical toxicity and that are compatible with integration into the drug discovery process. By combining SenzaGen’s technological expertise and our experience with inhaled compounds, we aim to develop a novel method to investigate whether novel drug-like molecules induce similar genomic signatures as known respiratory irritants. That could ultimately be applied to steer away from a potential irritancy hazard and enable the discovery of safer drugs for patients’” says Jorrit Hornberg, Director of Discovery Safety at AstraZeneca Gothenburg.
With this new agreement SenzaGen is initiating yet another important development collaboration with the pharmaceutical industry. SenzaGen’s focus is on introducing its tests into the pharmaceutical industry and on striving to demonstrate that there are safer and more accurate tests than those that use animals.
“This collaboration with AstraZeneca is a milestone for SenzaGen. It shows that leading pharmaceutical companies see the potential of next-generation testing and that this testing has considerable industrial value. Furthermore, the collaboration will provide SenzaGen with valuable knowledge and will help to refine and develop our GARD test for further commercial opportunities,”says Anki Malmborg Hager, CEO of SenzaGen.
The research project will run for 12 to 18 months and is in collaboration with AstraZeneca Gothenburg, contributing to the EU Horizon 2020 project granted to SenzaGen for the development of in vitro respiratory sensitization test.
For more information: Anki Malmborg Hager, CEO, SenzaGen AB
E-mail: amh@senzagen.com
Phone: 46 768 284822
About GARD GARD is a group of tests for assessing chemical skin sensitizers. The tests make use of genetic biomarkers for more than 200 genes which cover the entire immune reaction and are relevant to predicting the risk of hypersensitivity. The tests have 90% reliability. This compares with the current predominant test method, experiments on mice, which has a reliability rating of 72%. SenzaGen’s tests are also capable of measuring the potency of a substance’s allergenic properties. Consequently GARD tests provide a much more comprehensive basis for determining whether a substance should be classified as an allergen than current testing methods.
About SenzaGen SenzaGen makes it possible to replace animal experiments with in vitro genetic testing to determine the allergenicity of the chemicals we come into contact with in our daily lives, such as for example in cosmetics, pharmaceuticals, food products and dyes. The company’s patented tests are the most reliable on the market and provide more information than traditional evaluation methods. We ourselves sell the tests in Sweden and the USA, and we sell through partners in several other countries. Over the next few years the company will expand geographically, make alliances with more distribution partners and launch further unique tests. SenzaGen has its headquarters in Lund in Sweden and a subsidiary in San Francisco, USA. For more information visit www.senzagen.com.
About AstraZeneca AstraZeneca is a global, science-led biopharmaceutical company that focuses on the discovery, development and commercialisation of prescription medicines, primarily for the treatment of diseases in three main therapy areas – Oncology, Cardiovascular & Metabolic Diseases and Respiratory. The Company also is selectively active in the areas of autoimmunity, neuroscience and infection. AstraZeneca operates in over 100 countries and its innovative medicines are used by millions of patients worldwide.
The meeting promises more than 150 scientific sessions, approximately 350 ToxExpo exhibitors offering you the latest information on services and technology, thousands of abstract presentations, continuing education courses, awards presentations, receptions, career guidance and support, and more.
Presentations details
– Replacement of Animal Testing for CLP/GHS Classification of Skin Sensitizers is now possible using a Modified Genomic GARDskin [OECD TGP 4.106] Assay
SenzaGen presents the latest development towards reliable potency classification of chemicals according to CLP 1A and 1B, taking both LLNA and Human potency data in consideration. The assay is based on GARDskin and utilizes a refined gene expression signature developed specifically for potency categorization with high predictability.
Date: 3/13 Time: 13:30 -14:30 Room: 338
– Advantages with Genome Testing Opening up the Landscape for New Application Possibilities for Sensitization Testing using SenzaGen’s Genomic GARD Assay
SenzaGen’s GARD assay is based on expression analysis of predictive genomic biomarker signatures. Prediction calls of test substances are generated by computational methods based on machine learning. SenzaGen presents their experience in skin and respiratory sensitization testing, working with challenging compounds and mixtures, active substances, potency classification and NOEL interpretation.
Date: 3/14 Time: 13:30-14:30 Room: 338
These session are Exhibitor-Hosted Session. Although not an official part of the SOT Annual Meeting scientific program, its presentation is permitted by the Society.
Attendees are welcomed from researcher community, industry, manufacturers, regulatory agencies, consultants, CROs and every one interested in safety testing of chemical compounds.
https://senzagen.com/wp-content/uploads/2017/03/2_Time_Linkedin_1200x6282.png6281200admin_senzagenhttps://senzagen.com/wp-content/uploads/2017/01/LoggaSquare_RGB_liten_HF.pngadmin_senzagen2017-03-06 08:00:382017-07-18 15:50:52SenzaGen is presenting at hosted sessions at Society of Toxicology 56th Annual Meeting in Baltimore March 12-16, 2017
SenzaGen has received a grant of € 2,4 million from the EU’s research and innovation framework program Horizon 2020 to develop GARDair, the first generation test for respiratory allergies.
GARDair is the first in vitro test that can detect air-borne chemicals that cause respiratory allergies, by using a unique gene technology. This follows the international 3Rs principle, Reduce, Replace and Refine, and is the first cell-based alternative to animal testing for the respiratory tract. The grant of 2,4 million Euros means that SenzaGen can develop and refine GARDair at the same time as being able to increase the pace of commercial development and the launch of another new product line. The grant runs for 24 months.
-We are incredibly delighted and proud to be part of the EU’s investment in the future, Horizon 2020. The fact that they choose to invest in 3Rs technologies shows how important this is. It is a confirmation of the potential of our technology platform, as well as a unique business opportunity to build on our product portfolio of commercially viable tests at a significantly faster rate, says Anki Malmborg, CEO of SenzaGen.
-It is important in many different industries to be able to test the capacity of chemicals to affect the respiratory system, and today there are no tests for this. There’s a huge need for safety testing, and the grant from the EU gives us a unique opportunity together with partners from various different industries to develop the respiratory test of the future in accordance with OECD guidelines, says Henrik Johansson, Senior Scientist at SenzaGen. SenzaGen’s EU application has been coordinated by GAEU Consulting AB and received a exceptionally high score of 14,26 out of 15 in the EU evaluation, proving excellency both in the invention and in the application document.
Horizon 2020 in brief Horizon 2020 is the EU’s framework program for research and innovation. The program is the world’s largest investment in research and innovation and has a total budget of around € 80 billion (2014-2020). The aim is to ensure the EU’s global competitiveness. For more on Horizon 2020, see: https://ec.europa.eu/programmes/horizon2020/
For more information: Anki Malmborg Hager, CEO, SenzaGen AB
E-mail: amh@senzagen.com
Phone: +46 768 284822
About GARD By analyzing 200 and 389 markers, depending on the test, GARD generates massive amounts of data and delivers results with 90 % prediction accuracy. This can be compared to the golden standard, animal tests on mice, that provides 72 % prediction. SenzaGen’s test also has the ability to measure potency (strength) of a substance and can thus determine the degree to which a substance is an allergen.
About SenzaGen
SenzaGen provides dermal and respiratory in vitro testing for the cosmetic, chemical and pharmaceutical industries replacing the need for animal testing. The company’s unique test GARD is based on research from the Department of Immune Technology at Lund University. SenzaGen is based at Medicon Village in Lund, Sweden.
https://senzagen.com/wp-content/uploads/2017/03/Horizon-2020_400x400pxl.jpg296400Malin Bergholmhttps://senzagen.com/wp-content/uploads/2017/01/LoggaSquare_RGB_liten_HF.pngMalin Bergholm2016-11-29 06:02:002017-10-23 13:48:44SenzaGen awarded EU grant of € 2,4 million in heavy competition, to develop GARDair for industry use
