Kathrin S. Zeller, Andy Forreryd, Tim Lindberg, Ann-Sofie Albrekt, Aakash Chawade, Malin Lindstedt
Dept. of Immunotechnology, Lund University, Lund, Sweden; Swedish University of Agricultural Sciences, Alnarp, Sweden
Summary
The GARD assay is a cell-based transcriptional biomarker assay for the prediction of chemical ensitizers1 targeting key event 3, dendritic cell activation, of the skin sensitization AOP. Here, we present a modified assay based on Random Forest modelling, which is capable of predicting CLP
potency classes (1A – strong sensitizers, 1B – weak sensitizers, no category – non-sensitizers) as described by the European CLP regulation with an accuracy of 75 % (no cat), 75 % (1B) and 88 % (1A) based on a test set consisting of 18 chemicals previously unseen to the model.
We further can link the activation of distinct pathways to the chemical protein reactivity, showing that our transcriptomic approach can reveal information contributing to the understanding of underlying mechanisms in sensitization.
Results and Discussion
We here present a potency prediction approach based on a Random Forest model and 18 transcripts. 18 chemicals previously unseen to the model were classified as shown in Tables 1, 4 and Fig. 1. Interestingly, diethyl maleate, misclassified as 1A instead of 1B, is a human potency class 2 according to4, and iodopropynyl butylcarbamate, wrongly predicted as 1B instead of 1A, is classified as human potency class 44. Thus, the model seems to show more agreement with human data than CLP classifications (mainly derived from animal data) based on this limited dataset. Also Fig. 1C supports the hypothesis, that both data and model contain information allowing the prediction of human potency.
Furthermore, Key Pathway Advisor analysis reveals that these data can be used to investigate the cellular response in more detail (Table 3). In conclusion, we show that the modified GARD assay is capable of providing potency information, which is imperative for quantitative risk assessment of chemical sensitizers.
The GARD assay for potency assessment of skin sensitizing chemicals_ESTIV 2016_Zeller_p
SenzaGen introduces GARDpotency – a unique method for measuring the allergenicity of chemical substances
SenzaGen AB announces today the start of pilot sales of GARDpotency, the first animal-free method of analysis that can provide invaluable information on the allergenic potency of a chemical substance. GARDpotency is a unique complement to the previously launched test method GARDskin.
For a long time there has been huge demand from government and industry to be able to quantify the strength of chemicals’ allergenicity. SenzaGen’s new in vitro test method GARDpotency makes this possible for the first time. Quantitative information is a requirement of REACH, the EU chemicals regulation, and is of great help to companies that develop new cosmetics, pharmaceuticals and food products. Until now all testing for potency classification has been carried out on animals.
As a first step in the launch of GARDpotency, SenzaGen will present the test method for potential customers in connection with the Society of Toxicology’s annual conference 12-16 March 2017 in Baltimore, USA. The technology behind GARDpotency has been developed by a research group at Lund University. At the conference, which brings together around 6,500 toxicologists from more than 60 countries, the research team will present two scientific papers describing GARDpotency as well as new application areas for GARDskin. Summaries of the research results will soon be made available and will also be available on the SenzaGen website after the scientific publications are published.
“We see great market potential for GARDpotency. It is already possible to determine if a chemical can cause allergies, but there is a significant additional need among both manufacturers and regulatory bodies to be able to quantify how strong the allergenic effect is. Until now no animal-free testing has been available and we are extremely proud to be the first to offer this to the chemicals industry,” says SenzaGen’s CEO, Anki Malmborg Hager.
GARDskin is scientifically validated and has been approved for validation in accordance with OECD requirements on allergy tests. The test provides greater than 90 percent accuracy in the classification of allergenic substances.
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
Press release in pdf
SenzaGen introducerar GARDpotency – en unik möjlighet att kvantifiera kemiska ämnens allergiframkallande egenskaper
SenzaGen AB meddelar idag att bolaget inleder pilotförsäljning av GARDpotency – den första djurfria analysmetoden som kan ge värdefull information om den allergiframkallande styrkan för en kemisk substans. GARDpotency är ett unikt komplement till den redan lanserade testmetoden GARDskin.
Det har länge funnits ett starkt önskemål från myndigheter och industrin att kunna kvantifiera styrkan av kemikaliers allergiframkallande förmåga. SenzaGens nya provrörsbaserade testmetod GARDpotency ger för första gången just denna möjlighet. Kvantitativ information är ett krav i den europeiska kemikalieförordningen REACH och är till stor hjälp för företag som utvecklar nya kosmetika, läkemedel och livsmedel. Hittills har all testning för så kallad potency-klassificering gjorts på djur.
Som ett första steg i lanseringen av GARDpotency kommer SenzaGen att introducera testmetoden för potentiella kunder i samband med Society of Toxicologys årliga konferens den 12–16 mars 2017 i Baltimore, USA. Teknologin bakom GARDpotency har utvecklats av en forskargrupp vid Lunds Universitet. Forskargruppen kommer presentera två vetenskapliga arbeten vilka beskriver GARDpotency och nya applikationsområden för GARDskin vid konferensen, som samlar omkring 6 500 toxikologer från mer än 60 länder. Sammanfattningar av forskningsresultaten kommer göras tillgängliga inom kort och även publiceras på SenzaGens hemsida efter den vetenskapliga publikationen.
”Vi ser en stor marknadspotential för GARDpotency. Det är redan idag möjligt att bedöma huruvida ett kemiskt ämne kan framkalla allergier, men det finns ett betydande behov bland såväl tillverkare som myndigheter av att dessutom kunna kvantifiera hur stark den allergiframkallande effekten är. Det har inte tidigare funnits något djurfritt test att tillgå och vi är oerhört stolta över att vara först med att kunna erbjuda kemikaliebranschen ett sådant ”, säger SenzaGens vd, Anki Malmborg Hager.
GARDskin är vetenskapligt validerat och har godkänts för validering mot OECD:s krav på allergitester. Testet har mer än 90 procents träffsäkerhet i klassificeringen av allergiframkallande substanser.
För mer information
Anki Malmborg Hager, VD, SenzaGen AB
E-mail: amh@senzagen.com
Telefon: 0768 284822
Om GARD
GARD är provrörsbaserade tester för bedömning av kemiska hudsensibiliserare. Testerna utnyttjar genetiska biomarkörer för fler än 200 gener som täcker hela immunreaktionen och är relevanta för att förutsäga risken för överkänslighet. Testerna har en tillförlitlighet på 90%. Detta kan jämföras med den idag dominerande testmetoden, djurförsök på möss, vilken ger en tillförlitlighet på 72%. SenzaGens tester har också förmågan att mäta potensen (styrkan) av ett ämnes allergiframkallande egenskaper. GARD-testerna levererar därmed ett mer heltäckande underlag
Om SenzaGen
SenzaGen gör det möjligt att ersätta djurförsök med genetiska tester i provrör för att bedöma om de kemikalier vi kommer i kontakt med i vår vardag är allergiframkallande. Det kan till exempel handla om kosmetika, läkemedel, livsmedel och färgämnen. Bolagets patentskyddade tester är de mest tillförlitliga på marknaden och ger mer information än traditionella utvärderingsmetoder. Testerna säljs i egen regi i Sverige och USA, och genom partners i flera andra länder. De närmaste åren kommer bolaget expandera geografiskt, knyta till sig fler distributionspartners och lansera nya, unika tester. SenzaGen har sitt huvudkontor i Lund och dotterbolag i San Francisco, USA. För mer information, besök www.senzagen.com
Pressmeddelande som pdf
SenzaGen is presenting at hosted sessions at Society of Toxicology 56th Annual Meeting in Baltimore March 12-16, 2017
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.
SenzaGen presenterar på Unicorn Summit den 7 mars 2017
SenzaGens vd Anki Malmborg Hager kommer att presentera bolagets spännande utveckling vid Vator Securities Unicorn Summit den 7 mars 2017.
Presentationen kommer beskriva hur företaget kommersialiserar och vidareutvecklar sina genombaserade tester (GARD), vilka kan bedöma om kemikalier vi kommer i kontakt med i vår vardag är allergiframkallande – ett ständigt ökande problem. Testerna bygger på hur 200 gener i mänskliga celler reagerar på det ämnen som testas,vilket innebär att de är betydligt mer tillförlitliga och ger mer information än dagens analysmetoder och kan minska behovet av försöksdjur.
– Vårt test har en tillförlitlighet på 90 procent, medan djurförsök hamnade runt 75 procent, konstaterar Anki Mamborg Hager, vd för Senzagen.
Unicorn Summit är en kapitalmarknadsdag som anordnas av Vator Sercurities för att sammanföra några av de mest innovativa och snabbväxande svenska life science-bolagen med investerare. För mer information och anmälan till arrangemanget, se www.vatorsecurities.se
För mer information
Anki Malmborg Hager, VD, SenzaGen AB
E-mail: amh@senzagen.com
Telefon: 46 768 284822
SenzaGen gör det möjligt att ersätta djurförsök med genetiska tester i provrör för att bedöma om de kemikalier vi kommer i kontakt med i vår vardag är allergiframkallande. Det kan till exempel handla om kosmetika, läkemedel, livsmedel och färgämnen. Bolagets patentskyddade tester är de mest tillförlitliga på marknaden och ger mer information än traditionella utvärderingsmetoder. Testerna säljs i egen regi i Sverige och USA, och genom partners i flera andra länder. De närmaste åren kommer bolaget expandera geografiskt, knyta till sig fler distributionspartners och lansera nya, unika tester. SenzaGen har sitt huvudkontor i Lund och dotterbolag i San Francisco, USA. För mer information, besök www.senzagen.com
Evaluation of the GARD assay in a blind Cosmetics Europe study
ALTEX Online first published February 17, 2017 https://doi.org/10.14573/altex.1701121
Johansson H., Gradin R., Forreryd A., Agemark M., Zeller K., Johansson A., Larne O., van Vliet E., Borrebaeck C., Lindstedt M.,
Summary
Chemical hypersensitivity is an immunological response towards foreign substances, commonly referred to as sensitizers, which gives rise primarily to the clinical symptoms known as allergic contact dermatitis. For the purpose of mitigating risks associated with consumer products, chemicals are screened for sensitizing effects. Historically, such predictive screenings have been performed using animal models. However, due to industrial and regulatory demand, animal models for the purpose of sensitization assessment are being replaced by animalfree testing methods, a global trend that is spreading across industries and market segments. To meet this demand, the Genomic Allergen Rapid Detection (GARD) assay was developed. GARD is a novel, cell-based assay that utilizes the innate recognition of xenobiotic substances by dendritic cells, as measured by a multivariate readout of genomic biomarkers. Following cellular stimulation, chemicals are classified as sensitizers or non-sensitizers based on induced transcriptional profiles. Recently, a number of animal-free methods were comparatively evaluated by Cosmetic Europe, using a coherent and blinded test panel of reference chemicals with human and local lymph node assay data, comprising a wide range of sensitizers and non-sensitizers. In this paper, the outcome of the GARD assay is presented. It was demonstrated that GARD is a highly functional assay with a predictive performance of 83% in this Cosmetics Europe dataset. The average accumulated predictive accuracy of GARD across independent datasets was 86%, for skin sensitization hazard. Keywords: GARD, sensitization, in vitro, predictive accuracy, alternative methods
Link to article e-pub ahead of print
Testing Human Skin and Respiratory Sensitizers—What Is Good Enough?
Int. J. Mol. Sci. 2017, 18(2), 241; doi:10.3390/ijms18020241
Malmborg A., Borrebaeck C. A.K.
Abstract
genomics; skin sensitization; adverse outcome pathways; next generation in vitro tests
Podcast – interview with CEO, Anki Malmborg Hager and professor Carl Borrebaeck
SenzaGen awarded EU grant of € 2,4 million in heavy competition, to develop GARDair for industry use
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.
Press release in pdf
SenzaGen erhåller EU-anslag på 24 miljoner i hård konkurrens, för utveckling av GARDair för industrin
SenzaGen har fått ett anslag på 24 miljoner kronor av EU:s forsknings- och innovations ramprogram Horizon 2020 för att vidareutveckla första generationens test för luftvägsallergier – GARDair.
GARD
air är det första in vitro-testet som kan detektera luftburna kemikalier, som orsakar luftvägsallergier, genom en unik genomteknik. Detta följer den internationella 3R-principen, Reduce, Replace and Refine, och är det första cellbaserade alternativet till djurtester inom luftvägar. Anslaget om 24 miljoner innebär att SenzaGen kan utveckla och förfina GARDair samtidigt som bolaget får möjlighet att öka tempot i den kommersiella utvecklingen och lanseringen av ännu en ny produktlinje. Anslaget löper under 24 månader.
– Vi är otroligt glada och stolta över att vara en del i EU:s framtidssatsning Horizon 2020. Att de väljer att satsa på 3Rtekniker visar på hur viktigt detta är. Det är en bekräftelse på vår teknologiplattforms potential, men inte minst en unik industriell möjlighet att bygga vidare på vår produktportfölj av kommersiellt gångbara tester i en väsentligt snabbare takt, säger Anki Malmborg, VD på SenzaGen.
Möjligheten att testa kemikaliers förmåga att påverka det respiratoriska systemet är viktigt i många olika industrier och idag finns inga tester för detta. Behovet av säkerhetstestning är stort och anslaget från EU ger oss en unik möjlighet tillsammans med partners från olika industrier att utveckla framtidens luftvägstest i enlighet med OECDs riktlinjer, säger Henrik Johansson, Senior Scientist på SenzaGen.
GAEU Consulting AB har koordinerat SenzaGens EU-ansökan som fick ett sällsynt högt betyg med 14,26 av 15 möjliga. Detta vittnar om en enorm styrka både i produktinnovationen och hur ansökan och projektet är formulerat.
Kort om Horizon 2020
Horizon 2020 är EU:s ramprogram för forskning och innovation. Programmet är världens största satsning på forskning och innovation och har en total budget på runt 80 miljarder Euro(2 014-2020). Syftet är bland annat att säkerställa EU:s globala konkurrenskraft. Mer om Horizon 2020, se https://ec.europa.eu/programmes/horizon2020/
För mer information
Anki Malmborg Hager, VD, SenzaGen AB
E-mail: amh@senzagen.com
Telefon: +46 768 284822
Om GARD
Genom att analysera 200 respektive 389 markörer beroende på test, genererar GARD stora mängder data och levererar resultat med 90 % tillförlitlighet. Detta kan jämföras med den gyllene standarden som är djurtester på möss som ger endast 72 % tillförlitlighet. SenzaGens test har därtill förmågan att mäta potensen (styrkan) av en substans och kan därmed avgöra till vilken grad en substans orsakar allergi.
Om SenzaGen
SenzaGen erbjuder in vitro-tester för hud och luftvägar till kosmetisk, kemisk och farmaceutisk industri som ersätter behovet av djurförsök. Bolagets unika test GARD är baserat på forskning från Institutionen för Immunteknologi vid Lunds Tekniska Högskola. SenzaGen är baserat på Medicon Village i Lund, Sverige
Pressmeddelande som pdf
The GARD™ assay for potency assessment of skin sensitizing chemicals
Kathrin S. Zeller, Andy Forreryd, Tim Lindberg, Ann-Sofie Albrekt, Aakash Chawade, Malin Lindstedt
Dept. of Immunotechnology, Lund University, Lund, Sweden; Swedish University of Agricultural Sciences, Alnarp, Sweden
Summary
The GARD assay is a cell-based transcriptional biomarker assay for the prediction of chemical ensitizers1 targeting key event 3, dendritic cell activation, of the skin sensitization AOP. Here, we present a modified assay based on Random Forest modelling, which is capable of predicting CLP
potency classes (1A – strong sensitizers, 1B – weak sensitizers, no category – non-sensitizers) as described by the European CLP regulation with an accuracy of 75 % (no cat), 75 % (1B) and 88 % (1A) based on a test set consisting of 18 chemicals previously unseen to the model.
We further can link the activation of distinct pathways to the chemical protein reactivity, showing that our transcriptomic approach can reveal information contributing to the understanding of underlying mechanisms in sensitization.
Results and Discussion
We here present a potency prediction approach based on a Random Forest model and 18 transcripts. 18 chemicals previously unseen to the model were classified as shown in Tables 1, 4 and Fig. 1. Interestingly, diethyl maleate, misclassified as 1A instead of 1B, is a human potency class 2 according to4, and iodopropynyl butylcarbamate, wrongly predicted as 1B instead of 1A, is classified as human potency class 44. Thus, the model seems to show more agreement with human data than CLP classifications (mainly derived from animal data) based on this limited dataset. Also Fig. 1C supports the hypothesis, that both data and model contain information allowing the prediction of human potency.
Furthermore, Key Pathway Advisor analysis reveals that these data can be used to investigate the cellular response in more detail (Table 3). In conclusion, we show that the modified GARD assay is capable of providing potency information, which is imperative for quantitative risk assessment of chemical sensitizers.
The GARD assay for potency assessment of skin sensitizing chemicals_ESTIV 2016_Zeller_p