September 2016
Volume 57, Issue 12
Open Access
ARVO Annual Meeting Abstract  |   September 2016
The effect of oncometabolite 2-hydroxyglutarate on the biological behavior of differentiated melanocytes and uveal melanoma cells
Author Affiliations & Notes
  • Cindy Weidmann
    Axe médecine régénératrice et Centre universitaire d’ophtalmologie-Recherche, Centre de Recherche du CHU de Québec, Quebec City, Quebec, Canada
  • Colm F. Quirke
    Axe médecine régénératrice et Centre universitaire d’ophtalmologie-Recherche, Centre de Recherche du CHU de Québec, Quebec City, Quebec, Canada
  • Christine Yao
    Axe médecine régénératrice et Centre universitaire d’ophtalmologie-Recherche, Centre de Recherche du CHU de Québec, Quebec City, Quebec, Canada
  • Carolyne Mary Lowry
    Département de médecine nucléaire et de radiobiologie, Université de Sherbrooke, Sherbrooke, Quebec, Canada
  • Jade Pomerleau
    Axe médecine régénératrice et Centre universitaire d’ophtalmologie-Recherche, Centre de Recherche du CHU de Québec, Quebec City, Quebec, Canada
  • J. Richard Wagner
    Département de médecine nucléaire et de radiobiologie, Université de Sherbrooke, Sherbrooke, Quebec, Canada
  • Solange Landreville
    Axe médecine régénératrice et Centre universitaire d’ophtalmologie-Recherche, Centre de Recherche du CHU de Québec, Quebec City, Quebec, Canada
    Département d'ophtalmologie, Université Laval, Quebec City, Quebec, Canada
  • Footnotes
    Commercial Relationships   Cindy Weidmann, None; Colm F. Quirke, None; Christine Yao, None; Carolyne Lowry, None; Jade Pomerleau, None; J. Richard Wagner, None; Solange Landreville, None
  • Footnotes
    Support  Fonds Merck-FMed de la Fondation de l'Université Laval, Fonds de recherche du Québec – Santé (FRQS), Fondation du CHU de Québec, Canada Foundation for Innovation
Investigative Ophthalmology & Visual Science September 2016, Vol.57, 4117. doi:
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      Cindy Weidmann, Colm F. Quirke, Christine Yao, Carolyne Mary Lowry, Jade Pomerleau, J. Richard Wagner, Solange Landreville; The effect of oncometabolite 2-hydroxyglutarate on the biological behavior of differentiated melanocytes and uveal melanoma cells. Invest. Ophthalmol. Vis. Sci. 2016;57(12):4117.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : Liver metastasis is a dreaded complication of uveal melanoma (UM). Ocular tumors with a propensity to metastasize are less differentiated than their non-metastatic counterparts. Cell differentiation and metastatic potential are controlled, among others, by the precise regulation of DNA methylation. Hydroxylation of 5-methylcytosine (5-mC) in 5-hydroxymethylcytosine (5-hmC) is a new demethylation mechanism mediated by Ten-eleven translocation 5-methylcytosine oxygenases (TETs) and isocitrate dehydrogenases (IDHs). The overall level of DNA hydroxymethylation in the genome of several cancers is significantly lower than that observed in the corresponding differentiated tissues and is correlated with metastatic stage. Our preliminary data suggest a similar 5-hmC loss in UM. About 10% of cutaneous melanomas harbor a mutation of IDH genes that leads to the production of the oncometabolite 2-hydroxyglutarate (2-HG), which inhibits TET enzymes. Our research project aims to understand the mechanisms that deregulate DNA hydroxymethylation in UM.

Methods : mRNA expression of DNA methylases and demethylases was determined in UM, choroid, and melanocytes by gene expression profiling. Mutations in these enzymes were screened by Sanger sequencing. Percentage of 5-hmC of total cytosines was quantified in DNA from UM, choroid, and melanocytes by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). Low grade UMs and melanocytes were treated with the TET inhibitor 2-HG before studying their state of differentiation using morphologic analyses, cell proliferation assays, as well as quantification of 5-hmC using ELISA.

Results : IDH1 and IDH2 transcripts were repressed in UM (melanocyte/UM ratio: 9.1 and 12.6, respectively). IDH1 or IDH2 mutations were not detected in UM cell lines screened so far. We measured a significant loss of DNA hydroxymethylation in UM genome compared to melanocytes and choroid (mean % on total cytosines: UM, 0.0035%; choroid, 0.185%; melanocytes, 0.0345%). 2-HG exposure increased proliferation and decreased melanocytic dendrite length in melanocytes.

Conclusions : Our study will yield novel information about the UM epigenome, and how it contributes to metastasis. Alterations of DNA methylation are reversible, and can thus be targeted with drugs.

This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.

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