Journal Article

Comprehensive Search for Genes Involved in Thalidomide Teratogenicity Using Early Differentiation Models of Human Induced Pluripotent Stem Cells: Potential Applications in Reproductive and Developmental Toxicity Testing

Yamamoto, Naoki, Kojima, Hajime, Shinke, Koudai, Hata, Yuko, Takimoto, Tetsuya, Saya, Hideyuki, Horie, Tetsuhiro, Sugihara, Eiji, Ishigaki, Yasuhito, Kato, Yu, Inaba, Takeshi, Sakamoto, Takuya, Nagano, Osamu, Hiramatsu, Noriko, Nagai, Noriaki

2025

Overview

Developmental toxicity testing is essential to identify substances that may harm embryonic development. This study aimed to establish a protocol for evaluating developmental toxicity using human induced pluripotent stem cells (iPSCs) by analyzing cellular activity and gene expression changes. Two ICH S5(R3) positive substances, valproic acid (VPA), which is a substance previously detected as positive by other test methods, and thalidomide (Thalido), were examined during early trichoderm differentiation without fetal bovine serum. RNA-seq analysis identified seven candidate genes, including TP63, associated with altered expression following exposure to VPA or Thalido. These genes were implicated in pathways related to tissue development, cell growth, and molecular interactions. While the assay effectively detected VPA and Thalido, its limitations include testing only soluble substances and focusing on early differentiation stages. Nevertheless, the protocol demonstrates potential for the classification and evaluation of emerging modality drugs based on physical properties such as solubility, polarity, and pH. Integration with AI analysis may enhance its capacity to uncover genetic variations and evaluate previously uncharacterized substances. This study provides a foundation for alternative developmental toxicity testing methods, with further refinements in the culture method expected to improve accuracy and applicability in regulatory toxicology.

• Publisher: MDPI AG,MDPI
• Subject: candidate genes / Cell culture / Cell differentiation / Cell Differentiation - drug effects / Cell Differentiation - genetics / Cytotoxicity / developmental toxicity / Drug development / Embryo cells / Embryogenesis / Enzymes / Fetuses / Gene expression / Genetic analysis / Genetic diversity / Humans / Induced Pluripotent Stem Cells - cytology / Induced Pluripotent Stem Cells - drug effects / Induced Pluripotent Stem Cells - metabolism / iPSCs / Methods / Pharmaceuticals / Pluripotency / Reproduction - drug effects / Reproduction - genetics / RNA-seq analysis / Stem cells / Teratogenesis - drug effects / Teratogenesis - genetics / Teratogenicity / Teratogens - toxicity / Thalidomide / Thalidomide - toxicity / Toxicity / Toxicity testing / Toxicity Tests - methods / Valproic acid / Valproic Acid - toxicity