title: Identification of thalidomide-specific transcriptomics and proteomics signatures during differentiation of human embryonic stem cells.
abstract: Embryonic development can be partially recapitulated in vitro by differentiating human embryonic stem cells (hESCs). Thalidomide is a developmental toxicant in vivo and acts in a species-dependent manner. Besides its therapeutic value, thalidomide also serves as a prototypical model to study teratogenecity. Although many in vivo and in vitro platforms have demonstrated its toxicity, only a few test systems accurately reflect human physiology. We used global gene expression and proteomics profiling (two dimensional electrophoresis (2DE) coupled with Tandem Mass spectrometry) to demonstrate hESC differentiation and thalidomide embryotoxicity/teratogenecity with clinically relevant dose(s). Proteome analysis showed loss of POU5F1 regulatory proteins PKM2 and RBM14 and an over expression of proteins involved in neuronal development (such as PAK2, PAFAH1B2 and PAFAH1B3) after 14 days of differentiation. The genomic and proteomic expression pattern demonstrated differential expression of limb, heart and embryonic development related transcription factors and biological processes. Moreover, this study uncovered novel possible mechanisms, such as the inhibition of RANBP1, that participate in the nucleocytoplasmic trafficking of proteins and inhibition of glutathione transferases (GSTA1, GSTA2), that protect the cell from secondary oxidative stress. As a proof of principle, we demonstrated that a combination of transcriptomics and proteomics, along with consistent differentiation of hESCs, enabled the detection of canonical and novel teratogenic intracellular mechanisms of thalidomide.
authors.affiliation: Center of Physiology and Pathophysiology, Institute of Neurophysiology, Cologne, Germany.
authors.forename: Kesavan
authors.initials: K
authors.lastname: Meganathan
authors.forename: Smita
authors.initials: S
authors.lastname: Jagtap
authors.forename: Vilas
authors.initials: V
authors.lastname: Wagh
authors.forename: Johannes
authors.initials: J
authors.lastname: Winkler
authors.forename: John Antonydas
authors.initials: JA
authors.lastname: Gaspar
authors.forename: Diana
authors.initials: D
authors.lastname: Hildebrand
authors.forename: Maria
authors.initials: M
authors.lastname: Trusch
authors.forename: Karola
authors.initials: K
authors.lastname: Lehmann
authors.forename: Jürgen
authors.initials: J
authors.lastname: Hescheler
authors.forename: Hartmut
authors.initials: H
authors.lastname: Schlüter
authors.forename: Agapios
authors.initials: A
authors.lastname: Sachinidis
mesh_terms: D002454:Cell Differentiation / Q000187:drug effects* / Q000235:genetics*; D004305:Dose-Response Relationship, Drug; D015536:Down-Regulation / Q000187:drug effects / Q000235:genetics; D047108:Embryonic Development / Q000187:drug effects / Q000235:genetics; D053595:Embryonic Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D005121:Extremities / Q000196:embryology; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D005982:Glutathione Transferase / Q000235:genetics / Q000378:metabolism; D006321:Heart / Q000187:drug effects / Q000196:embryology; D006801:Humans; D007700:Kinetics; D021381:Protein Transport / Q000187:drug effects; D040901:Proteomics; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D013792:Thalidomide / Q000494:pharmacology*; D013997:Time Factors; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D051153:Wnt Proteins / Q000235:genetics / Q000378:metabolism; D060449:Wnt Signaling Pathway / Q000187:drug effects / Q000235:genetics
chemical_list: D012333:RNA, Messenger; D014157:Transcription Factors; D051153:Wnt Proteins; D013792:Thalidomide; D005982:Glutathione Transferase