Investigation of the effects of phthalates on in vitro thyroid models with RNA-Seq and ATAC-Seq

Frontiers in Endocrinology

Endocrinology

Lorenzo Moroni¹  Daniel J. Carvalho²  Stefan Giselbrecht²  Florian Caiment³  Duncan Hauser³  Marta Nazzari³  Mírian Romitti⁴  Sabine Costagliola⁴ 

[1] Department of Complex Tissue Regeneration, MERLN Institute for Technology-Inspired Regenerative Medicine, Maastricht University, Maastricht, Netherlands;Department of Instructive Biomaterials Engineering, MERLN Institute for Technology-Inspired Regenerative Medicine, Maastricht University, Maastricht, Netherlands;Department of Toxicogenomics, GROW School for Oncology and Reproduction, Maastricht University, Maastricht, Netherlands;Institute of Interdisciplinary Research in Molecular Human Biology (IRIBHM), Université Libre de Bruxelles, Brussels, Belgium;

关键词: ATAC-seq;    Nthy-ori 3-1;    organoids;    phthalates;    RNA-seq;    thyroid;   

DOI  :  10.3389/fendo.2023.1200211

 received in 2023-04-04, accepted in 2023-08-08,  发布年份 2023

来源: Frontiers

【 摘 要 】

IntroductionPhthalates are a class of endocrine-disrupting chemicals that have been shown to negatively correlate with thyroid hormone serum levels in humans and to cause a state of hyperactivity in the thyroid. However, their mechanism of action is not well described at the molecular level.MethodsWe analyzed the response of mouse thyroid organoids to the exposure to a biologically relevant dose range of the phthalates bis(2-ethylhexyl) phthalate (DEHP), di-iso-decylphthalate (DIDP), di-iso-nonylphthalate (DINP), and di-n-octylphthalate (DnOP) for 24 h and simultaneously analyzed mRNA and miRNA expression via RNA sequencing. Using the expression data, we performed differential expression and gene set enrichment analysis. We also exposed the human thyroid follicular epithelial cell line Nthy-ori 3-1 to 1 µM of DEHP or DINP for 5 days and analyzed changes in chromatin accessibility via ATAC-Seq.ResultsDose-series analysis showed how the expression of several genes increased or decreased at the highest dose tested. As expected with the low dosing scheme, the compounds induced a modest response on the transcriptome, as we identified changes in only mmu-miR-143-3p after DINP treatment and very few differentially expressed genes. No effect was observed on thyroid markers. Ing5, a component of histones H3 and H4 acetylation complexes, was consistently upregulated in three out of four conditions compared to control, and we observed a partial overlap among the genes differentially expressed by the treatments. Gene set enrichment analysis showed enrichment in the treatment samples of the fatty acid metabolism pathway and in the control of pathways related to the receptor signalling and extracellular matrix organization. ATAC-Seq analysis showed a general increase in accessibility compared to the control, however we did not identify significant changes in accessibility in the identified regions.DiscussionWith this work, we showed that despite having only a few differentially expressed genes, diverse analysis methods could be applied to retrieve relevant information on phthalates, showing the potential of in vitro thyroid-relevant systems for the analysis of endocrine disruptors.

【 授权许可】

Unknown   
Copyright © 2023 Nazzari, Romitti, Hauser, Carvalho, Giselbrecht, Moroni, Costagliola and Caiment