The Teratogenic Effects of Anti-Diabetic Drugs: In Vitro Assays Using Embryoid Body Morphogenesis

Abstract

Many adults take some form of medication to treat chronic conditions, however little is known about their effects on embryogenesis and pregnancy. With the increase in fertility problems society is facing and the increase in use of medications, it is important to understand what effects pharmaceuticals have on the developing embryo to ensure that future generations are not adversely affected by teratogenic compounds that can lead to birth defects. In this study, teratogenicity of anti-diabetic compounds was examined using the in vitro P19C5 embryoid body (EB) elongation morphogenesis model. As the EB model recapitulates key embryological events crucial for body patterning and axis formation, adverse impact of drugs on EB growth and elongation implicates their potential teratogenicity. Of the 31 anti-diabetic compounds examined, 27 decreased EB size and 11 either decreased or increased EB elongation at a given concentration. Morphogenetic impact of four specific anti-diabetic drugs, dapagliflozin, phenformin HCl, manidipine 2HCl, and resveratrol, was further examined by investigating expression patterns of key developmental regulators. Each drug exhibited a unique effect on gene expression patterns. Notably, dapagliflozin significantly reduced the level of the Wnt3 gene, a crucial regulator of the primitive streak formation. The Wnt-inhibiting effect of dapagliflozin was also confirmed by the TOPFLASH reporter assay. Furthermore, it was revealed that the drug impact on EB size was likely mediated by adverse effect on cell proliferation. These results suggest that anti-diabetic compounds impact EB morphogenesis by perturbing key developmental regulators, cellular signaling, and proliferation. The present research should lay the foundation for further investigations, including animal and human studies, to determine teratogenicity of specific anti-diabetes medications.