The effect of lead during the Flint water crisis on mouse embryonic stem cells self-renewal and differentiation markers

Abstract

During the Flint water crisis, the residents of Flint, Michigan experienced a significant increase in blood lead levels. For some this resulted in an increase as high as 40 μg/dL from 5 μg/dL, which is considered to be safe by the Center for Disease Control and Prevention. Since the extent of the effect of the lead exposure in early embryonic development is not greatly investigated, the aim of this study is to explore the effect of lead exposure at concentrations present in Flint, MI during the Flint water crisis in the embryonic development. The expression of pluripotency and self-renewal markers (Oct4, Sox2, Nanog and Zfp-42) coupled with morphological and alkaline phosphatase assays revealed that mouse embryonic stem cells (mESC) pluripotency and self-renewal capabilities are perturbed following exposure in a lead acetate concentration dependent manner. Moreover, mouse embryoid bodies (mEB), which provide ideal models for testing toxicity in vitro, revealed that lead acetate exposure induces fewer but larger mEBs, whereas gene expression analysis of lineage specific transcription factors showed an increased mRNA level of endodermal (Gata 4, Gata 6, Sox 7) and mesodermal markers (Eomes, Hand 1, Slug 1) while the mRNA level of ectodermal markers (Otx 2, Noggin, Sox 1) decreased. Taken all together, these results indicate that lead acetate disturbs the pluripotency of mESC and differentiation potential of mEBs by inhibiting differentiation towards ectodermal lineages and inducing it towards endodermal and mesodermal lineages.