Abstract
Triphenyl phosphate (TPhP) is an environmental PPARγ ligand, and growing evidence suggests that it is a metabolic disruptor. We have shown previously that the structurally similar ligand, tributyltin, does not induce brite adipocyte gene expression. Here, using in vivo and in vitro models, we tested the hypothesis that TPhP is a selective PPARγ ligand, which fails to induce brite adipogenesis. C57BL/6 J male mice were fed either a low or very high-fat diet for 13 weeks. From weeks 7–13, mice were injected intraperitoneally, daily, with vehicle, rosiglitazone (Rosi), or TPhP (10 mg/kg). Compared to Rosi, TPhP did not induce expression of browning-related genes (e.g. Elovl3, Cidea, Acaa2, CoxIV) in mature adipocytes isolated from inguinal adipose. To determine if this resulted from an effect directly on the adipocytes, 3T3-L1 cells and primary human preadipocytes were differentiated into adipocytes in the presence of Rosi or TPhP. Rosi, but not TPhP, induced expression of brite adipocyte genes, mitochondrial biogenesis and cellular respiration. Further, Rosi and TPhP-induced distinct proteomes and phosphoproteomes; Rosi enriched more regulatory pathways related to fatty acid oxidation and mitochondrial proteins. We assessed the role of phosphorylation of PPARγ in these differences in 3T3-L1 cells. Only Rosi protected PPARγ from phosphorylation at Ser273. TPhP gained the ability to stimulate brite adipocyte gene expression in the presence of the CDK5 inhibitor and in 3T3-L1 cells expressing alanine at position 273. We conclude that TPhP is a selective PPARγ modulator that fails to protect PPARγ from phosphorylation at ser273.
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Acknowledgements
This work was supported by the National Institute of Environmental Health Sciences Superfund Research Program P42 ES007381 (Jennifer Schlezinger), the National Institute of Diabetes and Digestive and Kidney Diseases R01 DK117161(Stephen Farmer) and the American Heart Association 17POST33660875 (Nabil Rabhi).
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This work was supported by the National Institute of Environmental Health Sciences Superfund Research Program P42 ES007381 to JJS, the National Institute of Diabetes and Digestive and Kidney Diseases R01DK117161 to SF and the American Heart Association 17POST33660875 to NR.
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JJS, SK, NR and SF contributed to the study conception and design. Experiments, data collection and analyses were performed by SK, NR, BCB, RH, and KW. Data analysis and interpretation were performed by all authors. The first draft of the manuscript was written by Stephanie Kim. All authors read, commented on, and approved the manuscript.
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Animal studies were reviewed and approved by the Institutional Animal Care and Use Committee at Boston University and performed in an American Association for the Accreditation of Laboratory Animal Care accredited facility (Animal Welfare Assurance Number: A3316-01). All animals were treated humanely and with regard for alleviation of suffering.
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Kim, S., Rabhi, N., Blum, B.C. et al. Triphenyl phosphate is a selective PPARγ modulator that does not induce brite adipogenesis in vitro and in vivo. Arch Toxicol 94, 3087–3103 (2020). https://doi.org/10.1007/s00204-020-02815-1
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DOI: https://doi.org/10.1007/s00204-020-02815-1