Inactivation of SDH and FH cause loss of 5hmC and increased H3K9me3 in paraganglioma/pheochromocytoma and smooth muscle tumors.

Abstract

Succinate dehydrogenase (SDH) and fumarate hydratase (FH) are tricarboxylic acid (TCA) cycle enzymes and tumor suppressors. Loss-of-function mutations give rise to hereditary paragangliomas/pheochromocytomas and hereditary leiomyomatosis and renal cell carcinoma. Inactivation of SDH and FH results in an abnormal accumulation of their substrates succinate and fumarate, leading to inhibition of numerous α-ketoglutarate dependent dioxygenases, including histone demethylases and the ten-eleven-translocation (TET) family of 5-methylcytosine (5 mC) hydroxylases. To evaluate the distribution of DNA and histone methylation, we used immunohistochemistry to analyze the expression of 5 mC, 5-hydroxymethylcytosine (5 hmC), TET1, H3K4me3, H3K9me3, and H3K27me3 on tissue microarrays containing paragangliomas/pheochromocytomas (n = 134) and hereditary and sporadic smooth muscle tumors (n = 56) in comparison to their normal counterparts. Our results demonstrate distinct loss of 5 hmC in tumor cells in SDH- and FH-deficient tumors. Loss of 5 hmC in SDH-deficient tumors was associated with nuclear exclusion of TET1, a known regulator of 5 hmC levels. Moreover, increased methylation of H3K9me3 occurred predominantly in the chief cell component of SDH mutant tumors, while no changes were seen in H3K4me3 and H3K27me3, data supported by in vitro knockdown of SDH genes. We also show for the first time that FH-deficient smooth muscle tumors exhibit increased H3K9me3 methylation compared to wildtype tumors. Our findings reveal broadly similar patterns of epigenetic deregulation in both FH- and SDH-deficient tumors, suggesting that defects in genes of the TCA cycle result in common mechanisms of inhibition of histone and DNA demethylases.

• Authors: Attje S Hoekstra, Marieke A de Graaff, Inge H Briaire-de Bruijn, Cor Ras, Reza Maleki Seifar, Ivonne van Minderhout, Cees J Cornelisse, Pancras C W Hogendoorn, Martijn H Breuning, Johnny Suijker, Esther Korpershoek, Henricus P M Kunst, Norma Frizzell, Peter Devilee, Jean-Pierre Bayley, Judith V M G Bovée
• Journal: Oncotarget (Oncotarget) Vol. 6 Issue 36 Pg. 38777-88 (Nov 17 2015) ISSN: 1949-2553 [Electronic] United States
• PMID: 26472283 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Proto-Oncogene Proteins
• 5-hydroxymethylcytosine
• 5-Methylcytosine
• Cytosine
• Mixed Function Oxygenases
• TET1 protein, human
• Succinate Dehydrogenase
• Histone-Lysine N-Methyltransferase
• Fumarate Hydratase

Topics

• 5-Methylcytosine (analogs & derivatives)
• Adrenal Gland Neoplasms (enzymology, genetics)
• Cell Nucleus (metabolism)
• Cytosine (analogs & derivatives, metabolism)
• Fumarate Hydratase (deficiency, genetics, metabolism)
• Gene Silencing
• HEK293 Cells
• Histone-Lysine N-Methyltransferase (metabolism)
• Humans
• Immunohistochemistry
• Mixed Function Oxygenases (metabolism)
• Paraganglioma (enzymology, genetics, pathology)
• Pheochromocytoma (enzymology, genetics, pathology)
• Proto-Oncogene Proteins (metabolism)
• Smooth Muscle Tumor (enzymology, genetics)
• Succinate Dehydrogenase (deficiency, genetics, metabolism)