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Hoxa5 undergoes dynamic DNA methylation and transcriptional repression in the adipose tissue of mice exposed to high-fat diet

International Journal of Obesity volume 40, pages 929–937 (2016)Cite this article

Subjects

Abstract

Background/Objectives:

The genomic bases of the adipose tissue abnormalities induced by chronic positive calorie excess have been only partially elucidated. We adopted a genome-wide approach to directly test whether long-term high-fat diet (HFD) exposure affects the DNA methylation profile of the mouse adipose tissue and to identify the functional consequences of these changes.

Subjects/Methods:

We have used epididymal fat of mice fed either high-fat (HFD) or regular chow (STD) diet for 5 months and performed genome-wide DNA methylation analyses by methylated DNA immunoprecipitation sequencing (MeDIP-seq). Mouse Homeobox (Hox) Gene DNA Methylation PCR, RT-qPCR and bisulphite sequencing analyses were then performed.

Results:

Mice fed the HFD progressively expanded their adipose mass accompanied by a significant decrease in glucose tolerance (P<0.001) and insulin sensitivity (P<0.05). MeDIP-seq data analysis revealed a uniform distribution of differentially methylated regions (DMR) through the entire adipocyte genome, with a higher number of hypermethylated regions in HFD mice (P<0.005). This different methylation profile was accompanied by increased expression of the Dnmt3a DNA methyltransferase (Dnmt; P<0.05) and the methyl-CpG-binding domain protein Mbd3 (P<0.05) genes in HFD mice. Gene ontology analysis revealed that, in the HFD-treated mice, the Hox family of development genes was highly enriched in differentially methylated genes (P=0.008). To validate this finding, Hoxa5, which is implicated in fat tissue differentiation and remodeling, has been selected and analyzed by bisulphite sequencing, confirming hypermethylation in the adipose tissue from the HFD mice. Hoxa5 hypermethylation was associated with downregulation of Hoxa5 mRNA and protein expression. Feeding animals previously exposed to the HFD with a standard chow diet for two further months improved the metabolic phenotype of the animals, accompanied by return of Hoxa5 methylation and expression levels (P<0.05) to values similar to those of the control mice maintained under standard chow.

Conclusions:

HFD induces adipose tissue abnormalities accompanied by epigenetic changes at the Hoxa5 adipose tissue remodeling gene.

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Acknowledgements

LP is the recipient of the EFSD/Lilly Research Fellowship 2015. This work has been supported, in part, by the European Foundation for the Study of Diabetes (EFSD), by the Ministero dell’Università e della Ricerca Scientifica (grants PRIN and FIRB-MERIT, and PON 01_02460) and by the Società Italiana di Diabetologia (SID-FO.DI.RI). This work was also supported by the P.O.R. Campania FSE 2007-2013, Project CREMe.

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Author notes

  1. V Costa and G A Raciti: These authors equally contributed to this work.

Authors and Affiliations

  1. Dipartimento di Scienze Mediche e Traslazionali dell'Università di Napoli 'Federico II', Naples, Italy

    L Parrillo, G A Raciti, M Longo, R Spinelli, C Nigro, V Vastolo, A Desiderio, F Zatterale, P Formisano, C Miele & F Beguinot

  2. URT dell'Istituto di Endocrinologia e Oncologia Sperimentale 'Gaetano Salvatore', Consiglio Nazionale delle Ricerche, Naples, Italy

    L Parrillo, G A Raciti, M Longo, R Spinelli, C Nigro, V Vastolo, A Desiderio, F Zatterale, P Formisano, C Miele & F Beguinot

  3. Institute of Genetics and Biophysics 'Adriano Buzzati-Traverso', National Research Council, Naples, Italy

    V Costa, R Esposito & A Ciccodicola

  4. Department of Science and Technology, DiST, 'Parthenope' University of Naples, Centro Direzionale, Naples, Italy

    A Ciccodicola

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  1. L Parrillo
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Correspondence to F Beguinot.

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Parrillo, L., Costa, V., Raciti, G. et al. Hoxa5 undergoes dynamic DNA methylation and transcriptional repression in the adipose tissue of mice exposed to high-fat diet. Int J Obes 40, 929–937 (2016). https://doi.org/10.1038/ijo.2016.36

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