TY - JOUR

T1 - Human embryonic stem cell-derived test systems for developmental neurotoxicity

T2 - A transcriptomics approach

AU - Krug, Anne K.

AU - Kolde, Raivo

AU - Gaspar, John A.

AU - Rempel, Eugen

AU - Balmer, Nina V.

AU - Meganathan, Kesavan

AU - Vojnits, Kinga

AU - Baquié, Mathurin

AU - Waldmann, Tanja

AU - Ensenat-Waser, Roberto

AU - Jagtap, Smita

AU - Evans, Richard M.

AU - Julien, Stephanie

AU - Peterson, Hedi

AU - Zagoura, Dimitra

AU - Kadereit, Suzanne

AU - Gerhard, Daniel

AU - Sotiriadou, Isaia

AU - Heke, Michael

AU - Natarajan, Karthick

AU - Henry, Margit

AU - Winkler, Johannes

AU - Marchan, Rosemarie

AU - Stoppini, Luc

AU - Bosgra, Sieto

AU - Westerhout, Joost

AU - Verwei, Miriam

AU - Vilo, Jaak

AU - Kortenkamp, Andreas

AU - Hescheler, Jürgen

AU - Hothorn, Ludwig

AU - Bremer, Susanne

AU - Van Thriel, Christoph

AU - Krause, Karl Heinz

AU - Hengstler, Jan G.

AU - Rahnenführer, Jörg

AU - Leist, Marcel

AU - Sachinidis, Agapios

N1 - Funding Information: Acknowledgments This study was funded by the FP7 project ESNATS, the DFG and the Doerenkamp-Zbinden Foundation. We thank Sara Skogsater (ARTTIC) for excellent work in coordinating the ESNATS consortium and for her helpful and professional support of the current study. The great technical assistance of Marion Kapitza is gratefully acknowledged.

PY - 2012/11/21

Y1 - 2012/11/21

N2 - Developmental neurotoxicity (DNT) and many forms of reproductive toxicity (RT) often manifest themselves in functional deficits that are not necessarily based on cell death, but rather on minor changes relating to cell differentiation or communication. The fields of DNT/RT would greatly benefit from in vitro tests that allow the identification of toxicant-induced changes of the cellular proteostasis, or of its underlying transcriptome network. Therefore, the 'human embryonic stem cell (hESC)- derived novel alternative test systems (ESNATS)' European commission research project established RT tests based on defined differentiation protocols of hESC and their progeny. Valproic acid (VPA) and methylmercury (MeHg) were used as positive control compounds to address the following fundamental questions: (1) Does transcriptome analysis allow discrimination of the two compounds? (2) How does analysis of enriched transcription factor binding sites (TFBS) and of individual probe sets (PS) distinguish between test systems? (3) Can batch effects be controlled? (4) How many DNA microarrays are needed? (5) Is the highest non-cytotoxic concentration optimal and relevant for the study of transcriptome changes? VPA triggered vast transcriptional changes, whereas MeHg altered fewer transcripts. To attenuate batch effects, analysis has been focused on the 500 PS with highest variability. The test systems differed significantly in their responses (\20 % overlap). Moreover, within one test system, little overlap between the PS changed by the two compounds has been observed. However, using TFBS enrichment, a relatively large 'common response' to VPA and MeHg could be distinguished from 'compound-specific' responses. In conclusion, the ESNATS assay battery allows classification of human DNT/RT toxicants on the basis of their transcriptome profiles.

AB - Developmental neurotoxicity (DNT) and many forms of reproductive toxicity (RT) often manifest themselves in functional deficits that are not necessarily based on cell death, but rather on minor changes relating to cell differentiation or communication. The fields of DNT/RT would greatly benefit from in vitro tests that allow the identification of toxicant-induced changes of the cellular proteostasis, or of its underlying transcriptome network. Therefore, the 'human embryonic stem cell (hESC)- derived novel alternative test systems (ESNATS)' European commission research project established RT tests based on defined differentiation protocols of hESC and their progeny. Valproic acid (VPA) and methylmercury (MeHg) were used as positive control compounds to address the following fundamental questions: (1) Does transcriptome analysis allow discrimination of the two compounds? (2) How does analysis of enriched transcription factor binding sites (TFBS) and of individual probe sets (PS) distinguish between test systems? (3) Can batch effects be controlled? (4) How many DNA microarrays are needed? (5) Is the highest non-cytotoxic concentration optimal and relevant for the study of transcriptome changes? VPA triggered vast transcriptional changes, whereas MeHg altered fewer transcripts. To attenuate batch effects, analysis has been focused on the 500 PS with highest variability. The test systems differed significantly in their responses (\20 % overlap). Moreover, within one test system, little overlap between the PS changed by the two compounds has been observed. However, using TFBS enrichment, a relatively large 'common response' to VPA and MeHg could be distinguished from 'compound-specific' responses. In conclusion, the ESNATS assay battery allows classification of human DNT/RT toxicants on the basis of their transcriptome profiles.

KW - Alternative testing strategies

KW - Methylmercury

KW - Reproductive toxicity

KW - Transcription factor

KW - Valproic acid

UR - http://www.scopus.com/inward/record.url?scp=84872346334&partnerID=8YFLogxK

U2 - 10.1007/s00204-012-0967-3

DO - 10.1007/s00204-012-0967-3

M3 - Article

C2 - 23179753

AN - SCOPUS:84872346334

VL - 87

SP - 123

EP - 143

JO - Archives of toxicology

JF - Archives of toxicology

SN - 0340-5761

IS - 1

ER -