title: 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.affiliation: Department of Human Genetics, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Attje S
authors.initials: AS
authors.lastname: Hoekstra
authors.affiliation: Department of Pathology, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Marieke A
authors.initials: MA
authors.lastname: de Graaff
authors.affiliation: Department of Pathology, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Inge H
authors.initials: IH
authors.lastname: Briaire-de Bruijn
authors.affiliation: Department of Biotechnology, Delft University of Technology, Delft, The Netherlands.
authors.forename: Cor
authors.initials: C
authors.lastname: Ras
authors.affiliation: Department of Biotechnology, Delft University of Technology, Delft, The Netherlands.
authors.forename: Reza Maleki
authors.initials: RM
authors.lastname: Seifar
authors.affiliation: Department of Clinical Genetics, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Ivonne
authors.initials: I
authors.lastname: van Minderhout
authors.affiliation: Department of Pathology, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Cees J
authors.initials: CJ
authors.lastname: Cornelisse
authors.affiliation: Department of Pathology, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Pancras C W
authors.initials: PC
authors.lastname: Hogendoorn
authors.affiliation: Department of Clinical Genetics, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Martijn H
authors.initials: MH
authors.lastname: Breuning
authors.affiliation: Department of Pathology, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Johnny
authors.initials: J
authors.lastname: Suijker
authors.affiliation: Department of Pathology, Josephine Nefkens Institute, Erasmus Medical Center Rotterdam, Rotterdam, The Netherlands.
authors.forename: Esther
authors.initials: E
authors.lastname: Korpershoek
authors.affiliation: Department of Otorhinolaryngology, Head and Neck Surgery, Radboud University Nijmegen Medical Center, Nijmegen, The Netherlands.
authors.forename: Henricus P M
authors.initials: HP
authors.lastname: Kunst
authors.affiliation: Department of Pharmacology, Physiology & Neuroscience, School of Medicine, University of South Carolina, Columbia, SC, USA.
authors.forename: Norma
authors.initials: N
authors.lastname: Frizzell
authors.affiliation: Department of Human Genetics, Leiden University Medical Center, Leiden, The Netherlands.|Department of Pathology, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Peter
authors.initials: P
authors.lastname: Devilee
authors.affiliation: Department of Human Genetics, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Jean-Pierre
authors.initials: JP
authors.lastname: Bayley
authors.affiliation: Department of Pathology, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Judith V M G
authors.initials: JV
authors.lastname: Bovée
mesh_terms: D044503:5-Methylcytosine / Q000031:analogs & derivatives; D000310:Adrenal Gland Neoplasms / Q000201:enzymology / Q000235:genetics*; D002467:Cell Nucleus / Q000378:metabolism; D003596:Cytosine / Q000031:analogs & derivatives* / Q000378:metabolism; D005649:Fumarate Hydratase / Q000172:deficiency / Q000235:genetics* / Q000378:metabolism; D020868:Gene Silencing; D057809:HEK293 Cells; D011495:Histone-Lysine N-Methyltransferase / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry; D006899:Mixed Function Oxygenases / Q000378:metabolism; D010235:Paraganglioma / Q000201:enzymology / Q000235:genetics* / Q000473:pathology; D010673:Pheochromocytoma / Q000201:enzymology / Q000235:genetics* / Q000473:pathology; D011518:Proto-Oncogene Proteins / Q000378:metabolism; D018235:Smooth Muscle Tumor / Q000201:enzymology / Q000235:genetics*; D013385:Succinate Dehydrogenase / Q000172:deficiency / Q000235:genetics* / Q000378:metabolism
chemical_list: D011518:Proto-Oncogene Proteins; C011865:5-hydroxymethylcytosine; D044503:5-Methylcytosine; D003596:Cytosine; D006899:Mixed Function Oxygenases; C462539:TET1 protein, human; D013385:Succinate Dehydrogenase; D011495:Histone-Lysine N-Methyltransferase; D005649:Fumarate Hydratase
keywords: fumarate hydratase; hereditary leiomyomatosis and renal cell carcinoma; methylation; paraganglioma; succinate dehydrogenase