The Validation of GARD®skin

Sandberg P, Johansson A, Agemark M, Gradin R, Larne O, Appelgren H, Forreryd A, Jerre A, Edwards A, Hoepflinger V, Burleson F, Gehrke H, Roggen E, Johansson H
SenzaGen, Lund, Sweden, Burleson Research Technologies, Morrisville, US, Eurofins, Munich, Germany

Introduction
The prevalence of allergic contact dermatitis (ACD) is estimated to >20% in the western world. Not only the individual is affected, but downstream socioeconomic effects are high. To minimize exposure, chemicals must be safety tested. Traditional testing strategies like the murine local lymph node assay (LLNA) comprise animals, but the regulatory authorities, public opinion and economic interests require animal-free models. The Genomic Allergen Rapid Detection skin (GARD®skin) is an in vitro assay addressing this need. Here, we present the results of the GARD®skin ring trial (OECD TGP 4.106) for validity of the assay. In addition, we show data for GARD®potency – a complementary assay developed to categorize identified senitizers as CLP 1A or 1B.

 

Conclusions

Transfer study
Transferability: 100%

Validation study
Reproducibility
WLR: 82 – 89%BLR: 92% (92 – 100%)
Test performance
– Accuracy: 94%
– Sensitivity: 93%
– Specificity: 96%

A blinded ring trial was performed to assess the functionality of the GARDskin assay. The data demonstrates that GARDskin is a powerful tool for assessment of chemical skin sensitizers, with a predictive accuracy of 94% and excellent reproducibility between laboratories.
In addition, we show that GARDpotency accurately assesses potency of identified sensitizers.

Poster download

Predicting skin sensitizaers with confidence-Using conformal prediction to determine applicability domain

Objective
GARD – Genomic Allergen Rapid Detection – is a state of the art non-animal based technology platform for classification of skin sensitizing chemicals. The assay has proven to be reliable and highly accurate for identification of skin sensitizing chemicals, and consistently reports predictive performances > 90% across external test sets. The aim of the current project is to complement assessments of average model performance with an estimate of uncertainty involved in each individual prediction, thus allowing for classification of skin sensitizers with confidence.

Results
An Internal validation procedure was initially performed on samples in the GARD training set (n=38) using the strategy described in Fig. 3A. Results from this exercise is summarized in Fig. 4A. Conformal prediction by definition allows the user to determine a reasonable and acceptable significance level to guarantee a maximum error rate in predictions. The significance level was set to 15%, i.e. the model was allowed to make a maximum of 15% errors. Performance of the conformal predictor was measured by validity and efficiency. A model was valid if the number of prediction errors did not exceed the significance level, while efficiency corresponded to the percentage of single class predictions. Internal validation of the training data resulted in a valid and highly effective model (92% single classifications, 1 empty, 2 both), indicating that the ambitious significance level was at a reasonable level for the GARD® assay. Following internal validation, samples in a large external test set (n =70) was classified within the CP framework as described in Fig. 3B, which resulted in generation of a valid and highly efficient model (99% single classifications, 0 empty, 1 both) (Fig.4B). Additional data on model performance is illustrated in Table 1.

Poster download

The use of GARDskin for sensitization evaluation of cosmetic ingredients and ‘real-life’ mixtures

Renato Ivan de Ávila, Tim Lindbergh, Malin Lindstedt and Marize Campos Valadares
Lab. of Education and Research in Pharmacology and Cellular Toxicology, Faculty of Pharmacy, Federal University of Goiás, Goiânia, Goiás, Brazil.
Department of Immunotechnology, Medicon Village, Lund University, Lund, Sweden.

 

Introduction
Genomic fingerprints in dendritic cells after chemical exposure is a recent strategy in in vitro techniques for skin sensitization hazard. Within this perspective, Genomic Allergen Rapid Detection (GARDskinTM), an assay based on a support vector machine (SVM) model, was developed for identifying contact allergens using a myeloid cell line as a surrogate for dendritic cells. Predictive system behind the GARDskin™ consists on the transcriptional quantitative analysis of 200 genes, referred as the GARDskin™ prediction signature. Mechanistically, GARDskin™ is linked to key event 3 “Activation of DCs”, as defined by the Adverse Outcome Pathways for skin sensitization published in 2012 by OECD (https://read.oecd-ilibrary.org/environment/the-adverse-outcome-pathway-for-skin-sensitisation-initiated-by-covalent-binding-to-proteins_9789264221444-en#page1)

 

Resluts
Information declared on the label and lawsone and PDD levels found in ten commercial henna-based hair coloring cosmetics are show in Table 1. Since all products analyzed were declared as henna cosmetics by the manufactures, the presence of LAW, the main active phytochemical of henna, was then expected in all samples. However, HPLC analysis showed no LAW level in the product nº 2, suggesting falsification. Furthermore, the presence of PPD was declared on the products nº 2 and 8 only by the manufactures. However, this substance was detected in all products, suggesting undisclosed adulteration.

Poster:
The use of GARDskin for sensitization evaluation of cosmetic ingredients and ‘real-life’ mixtures

SenzaGen co-hosting international workshop on Skin Sensitization in Brazil 21st August 2018

SenzaGen is co-hosting a workshop on skin sensitization at the Federal University of Goiás, Faculty of Pharmacy, Goiás.

The workshop, relevant both for Industry and Academy, is a collaboration between SenzaGen AB – Laboratório de Farmacologia e Toxicologia Celular- FarmaTec-FF on new tools for skin sensitization testing. During the workshop you will have the chance to get acquainted with the GARD platform, learn about hands-on usage and learn how the GARD method was developed as well as how it can help you to test your product in a safe and secure way. Further, the workshop will give you information on the current situation for alternative methods in Brazil, the skin sensitization AOP and the regulatory framework.

For more information and invitation read the Faculty of Pharmacy webpage

SenzaGen presents in Brazil

SenzaGen is a proud sponsor and contributor at the 2nd  Pan-American Conference for Alternative Methods on August 23-24, 2018 in Rio de Janeiro.

We are very pleased to see the changes in Brazil and support the development on introducing alternative methods for chemical testing. The country is now banning animal tests in cosmetics in one state after another, and the progress to ban animal testing in cosmetics is of great importance in the whole Pan-American Region. This conference brings South, Central and North America together to further find alternatives to animal testing and build collaboration and exchange scientific ideas. It is organised by CAAT (Center for Alternatives to Animal Testing) and local organizers, Inmetro and Natura Cosmetics in Brazil and the focus for this conference is on the 6R’s: Replacement, Refinement, Reduction,  Read-across, Relevance and Roadmaps.

SenzaGen Scientist Dr. Andy Forreryd will hold a presentation on Friday August 24, at 11.50 AM in Room 1:
The GARD™ assay for skin sensitization testing – validation, predictive performance and applicability for cosmetic ingredients.

 

The GARD platform will also be presented in a poster by Renato Ivan de Ávila, PhD student at the Federal University of Goiás, Brazil:

The use of GARDskin™ for skin sensitization evaluation of cosmetic ingredients and “real-life” mixtures.
Poster no: 17

 

Get more information about the conference HERE
Look at the programme HERE

Prediction of chemical respiratory sensitizers using GARD

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.

Poster:

Prediction of chemical respiratory sensitizers using GARD_LIVe2018

SenzaGen to present at LIVe 2018 in Nice, July 5th-6th

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!

Link to conference website

Link to programme

 

Vi anställer! – Är du vår nya kollega?

Laboratorieingenjör

Vi behöver förstärka vårt team med en engagerad och strukturerad person som trivs med rutinarbete i en dynamisk verksamhet!

Vill du arbeta med produkter som förbättrar människors hälsa utan att använda djurförsök? Då är tjänsten som laboratorieingenjör hos oss något för dig!

Om SenzaGen

SenzaGen AB är ett publikt Biotech-bolag som erbjuder och utvecklar den nya generationens djurfria testmetoder för bedömning av immunotoxikologiska egenskaper hos bl a kemikalier och komplexa blandningar. SenzaGen AB bildades 2010 som ett spin-out bolag från Institutionen för Immunteknologi, Lunds Universitet. Idag har bolaget 20 anställda och våra kunder finns inom kosmetika-, kemikalie- och läkemedelsindustrin. SenzaGen har mottagit flertalet utmärkelser såsom plats på Ny Teknik och Affärsvärldens 33-lista två år i rad, PWCs SKAPA Innovationspris samt Dagens Industris Gasellsprånget. SenzaGen noterades på Nasdaq First North i september 2017 och vi finns på Medicon Village i Lund.

Om GARD

GARD (Genomic Allergen Rapid Detection) är en state-of-the-art in vitro-plattform som med hjälp av patenterade genomiska biomarkörsignaturer och maskininlärning bedömer kemiska substansers kapacitet att orsaka hud- eller luftvägsallergi. Testet är unikt då det efterliknar människans immunreaktion.

SenzaGens ledande produkt, GARDskin, är en vetenskapligt granskad analysmetod med en mycket hög noggrannhet på >90%. Tillsammans med vår nästkommande produkt, GARDpotency, genomgår GARDskin för närvarande en validering hos OECD och ECVAM.

Arbetsuppgifter

  • Delta i kund- och utvecklingsprojekt som laborativ resurs
  • Utföra laboratoriearbete enligt gällande kvalitetssystem och inom överenskomna tidsramar
  • Dokumentera och sammanställa analysdata
  • Köpa in förbrukningsmaterial till laboratoriet
  • Ha ansvar för den dagliga driften på laboratoriet
  • Aktivt deltaga i interna möten

Kvalifikationer

Du har en examen som biomedicinsk analytiker eller motsvarande med flera års erfarenhet av laboratoriearbete i cellbiologisk miljö och är van att genomföra analyser enligt metodbeskrivning. Kunskaper inom cellodling, flödescytometri och RNA-hantering är särskilt meriterande. Du besitter naturligt ett kvalitetstänk. Du är kommunikativ i svenska och engelska i tal och skrift samt är van att arbeta med Officepaketet. Som person är du strukturerad, noggrann och har stort fokus på att leverera arbete med hög kvalitet och i tid. Du känner ansvar och engagemang för din uppgift och du är flexibel och trivs att arbeta i en grupp där alla strävar mot samma mål. Då vi är ett litet sammansvetsat team lägger vi stor vikt på lämplighet och personlighet.

Vi erbjuder

Som anställd hos SenzaGen erbjuds du att arbeta i en kreativ och engagerande miljö och vara del i ett innovativt företag som ligger i frontlinjen inom in vitro-metodik och sensibilisering. Företaget befinner sig i en spännande fas där alla strävar mot att ta företaget och vår vision framåt! Tjänsten är en 6 månaders provanställning med mål att övergå i en tillsvidareanställning. Tjänsten är heltid (40 timmar/vecka) med placering hos SenzaGen på Medicon Village i Lund.

Ansökan

Du ansöker till tjänsten genom att e-posta ditt CV och personliga brev till ansokan@senzagen.com senast 2018-07-15. Vi hanterar ansökningarna löpande så vänligen skicka din ansökan snarast!

Kontaktperson är Maria Agemark (0707-713114 eller maria.agemark@senzagen.com) men notera att din ansökan behöver registreras hos ansokan@senzagen.com

ANSÖKAN SOM PDF

Seminar at the Swedish Parliamentary Animal Protection Forum in collaboration with Djurens Rätt (Animal Rights), The Body Shop and Cruelty Free International

On Wednesday May 23rd 2018, SenzaGen’s CEO Anki Malmborg Hager participated in a breakfast seminar at the Swedish Parliament organized by the Parliamentary Animal Protection Forum, Birgitta Ohlsson (L) and Jens Holm (V), in collaboration with Djurens Rätt (Animal Rights), The Body Shop and Cruelty Free International. The seminar was organized to provide on-site information about Swedish politicians’ views on a global ban of animal testing in cosmetics, and Anki gave a presentation on animal-free methods for the testing of chemicals and cosmetics.

A few weeks ago, the EU Parliament agreed (by 620 votes to 14) that the European Union should work towards a global ban of animal testing of cosmetic products. But in many parts of the world there are still no laws to ban animal testing of cosmetic products and ingredients. This despite the availability of approved and scientifically validated more efficient and accurate non-animal based methods that are both faster and often cheaper.

Cruelty Free International is currently running a worldwide campaign together with The Body Shop under the hashtag #Foreveragainstanimaltesting to put an end to animal testing globally, forever. There was considerable interest at this seminar in SenzaGen’s in vitro method, which is based on human cells instead of animals.

“The fact that cosmetics and chemicals are still tested on animals in large parts of the world is really hard to understand when new methods are much more accurate. On average, animal tests have an accuracy of about 72% and the test we developed, GARDskin, offers an accuracy of more than 90%. And apart from the difference in accuracy, we are getting answers to how the human immune system is affected rather than how mice are affected,” says Anki Malmborg Hager, SenzaGen’s CEO.

The seminar is part of a very important initiative to pursue the issue of animal experiments both politically and via opinion, and, above all, to demand safer testing of chemical ingredients in the products that surround us in everyday life.

“It has been a wonderful opportunity for us to participate in this seminar and we look forward to being able to continue contributing to the entire project in the best way we can,” says Anki Malmborg Hager, CEO, SenzaGen.

 

For more information, please contact:
Anki Malmborg Hager, CEO, SenzaGen AB
Email: anki.malmborg.hager@senzagen.com
Telephone: +46 768 284822

New opportunity to join a free presentation on Sensitization testing using GARD™

Don’t miss this new opportunity to join a free presentation on Sensitization testing using GARD™

“Sensitization testing and hazard assessment using functional genomics with GARD™”

A new presentation has been scheduled to Wed, May 30, 2018 1:00 PM – 2:00 PM EDT (19:00 CET)

This free webinar set for US time zone will give insight into the latest in vitro technology using human cell lines and genomics for skin and respiratory sensitization, potency classification CLP 1A and 1B of skin sensitizers and with ability to test complex mixtures. A thorough explanation of the GARD™ technology, how it compares to other in vitro skin sensitization assays and how it is a highly reliable method for safety assessment, all industries.
Sensitization testing based on functional genomics

The webinar forum gives you a good chance to ask questions after the presentation.

We are very much looking forward to meeting you virtually in the webinar platform!

Read more and register on our website in the Event calendar