| Valproic Acid Teratogenicity: A Toxicogenomics Approach Kim Kultima, Anna-Maja Nyström, Birger Scholz, Anne-Lee Gustafson,
Lennart Dencker, and Michael Stigson
Department of Pharmaceutical Biosciences, Division of Toxicology, The Biomedical
Center, Uppsala University, Uppsala, Sweden Abstract
Embryonic development is a highly coordinated set of processes that depend on hierarchies of signaling and gene regulatory networks, and the disruption of such networks may underlie many cases of chemically induced birth defects. The antiepileptic drug valproic acid (VPA) is a potent inducer of neural tube defects (NTDs) in human and mouse embryos. As with many other developmental toxicants however, the mechanism of VPA teratogenicity is unknown. Using microarray analysis, we compared the global gene expression responses to VPA in mouse embryos during the critical stages of teratogen action in vivo with those in cultured P19 embryocarcinoma cells in vitro. Among the identified VPA-responsive genes, some have been associated previously with NTDs or VPA effects [vinculin, metallothioneins 1 and 2 (Mt1, Mt2) , keratin 1-18 (Krt1-18) ], whereas others provide novel putative VPA targets, some of which are associated with processes relevant to neural tube formation and closure [transgelin 2 (Tagln2) , thyroid hormone receptor interacting protein 6, galectin-1 (Lgals1) , inhibitor of DNA binding 1 (Idb1) , fatty acid synthase (Fasn) , annexins A5 and A11 (Anxa5, Anxa11) ], or with VPA effects or known molecular actions of VPA (Lgals1, Mt1, Mt2, Id1, Fasn, Anxa5, Anxa11, Krt1-18) . A subset of genes with a transcriptional response to VPA that is similar in embryos and the cell model can be evaluated as potential biomarkers for VPA-induced teratogenicity that could be exploited directly in P19 cell-based in vitro assays. As several of the identified genes may be activated or repressed through a pathway of histone deacetylase (HDAC) inhibition and specificity protein 1 activation, our data support a role of HDAC as an important molecular target of VPA action in vivo. Key words: biomarker, embryocarcinoma, galectin-1, histone deacetylase, in vitro toxicology, metallothionein, microarray, mouse embryo, neural tube defect, Sp1, teratogen, valproic acid, vinculin. Environ Health Perspect 112:1225-1235 (2004) . doi:10.1289/txg.7034 available via http://dx.doi.org/ [Online 3 June 2004]
Address correspondence to M. Stigson, Department of Pharmaceutical Biosciences, Division of Toxicology, Uppsala University, BMC, Box 594, SE-75124 Uppsala, Sweden. Telephone: 46 18 4714208 or 46 730 216605. Fax: 46 18 4714253. E-mail: michael.stigson@farmbio.uu.se Supplemental material is available online (http://ehp.niehs.nih.gov/txg/
members/2004/7034/supplemental.pdf) . Microarray data are available in the ArrayExpress database (http://www.ebi.ac.uk/arrayexpress/) at accession numbers A-MEXP-32 and E-MEXP-45. We thank R. Engdahl and L. Norgren for excellent technical assistance, and J. Hansson, M. Norrman, E. Lopez Fernandez de Villaverde, and K. Zemiran for help. We also thank I. Lönnstedt and D. von Rosen for valuable discussions about statistical issues and data analysis. This work was supported by grant 02/04-36 from the Swedish Animal Welfare Agency (http://www.djurskyddsmyndigheten.se) . M.S was supported by a grant from AstraZeneca, and A-L.G. by the Swedish Medical Research Council (MFR) . The authors declare they have no competing financial interests. Received 16 February 2004 ; accepted 3 June 2004.
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