Background:
Osteoclasts are major actors in the maintenance of bone homeostasis. The full functional maturation of osteoclasts from monocyte lineage cells is essential for the degradation of old/damaged bone matrix. Diuron is one of the most frequently encountered herbicides, particularly in water sources. However, despite a reported delayed ossification
in vivo
, its impact on bone cells remains largely unknown.
Objectives:
The objectives of this study were to first better characterize osteoclastogenesis by identifying genes that drive the differentiation of
monocyte progenitors into osteoclasts and to evaluate the toxicity of diuron on osteoblastic and osteoclastic differentiation
in vitro
.
Methods:
We performed chromatin immunoprecipitation (ChIP) against H3K27ac followed by ChIP-sequencing (ChIP-Seq) and RNA-sequencing (RNA-Seq) at different stages of differentiation of
monocytes into active osteoclasts. Differentially activated super-enhancers and their potential target genes were identified. Then to evaluate the toxicity of diuron on osteoblasts and osteoclasts, we performed RNA-Seq and functional tests during
in vitro
osteoblastic and osteoclastic differentiation by exposing cells to different concentrations of diuron.
Results:
The combinatorial study of the epigenetic and transcriptional remodeling taking place during differentiation has revealed a very dynamic epigenetic profile that supports the expression of genes vital for osteoclast differentiation and function. In total, we identified 122 genes induced by dynamic super-enhancers at late days. Our data suggest that high concentration of diuron (
) affects viability of mesenchymal stem cells (MSCs)
in vitro
associated with a decrease of bone mineralization. At a lower concentration (
), an inhibitory effect was observed
in vitro
on the number of osteoclasts derived from
monocytes without affecting cell viability. Among the diuron-affected genes, our analysis suggests a significant enrichment of genes targeted by pro-differentiation super-enhancers, with an odds ratio of 5.12 (
).
Discussion:
Exposure to high concentrations of diuron decreased the viability of MSCs and could therefore affect osteoblastic differentiation and bone mineralization. This pesticide also disrupted osteoclasts maturation by impairing the expression of cell-identity determining genes. Indeed, at sublethal concentrations, differences in the expression of these key genes were mild during the course of
in vitro
osteoclast differentiation. Taken together our results suggest that high exposure levels of diuron could have an effect on bone homeostasis.
https://doi.org/10.1289/EHP11690
...