title: Aberrant succination of proteins in fumarate hydratase-deficient mice and HLRCC patients is a robust biomarker of mutation status.
abstract: Germline mutations in the FH gene encoding the Krebs cycle enzyme fumarate hydratase predispose to hereditary leiomyomatosis and renal cell cancer (HLRCC) syndrome. FH-deficient cells and tissues accumulate high levels of fumarate, which may act as an oncometabolite and contribute to tumourigenesis. A recently proposed role for fumarate in the covalent modification of cysteine residues to S-(2-succinyl) cysteine (2SC) (termed protein succination) prompted us to assess 2SC levels in our existing models of HLRCC. Herein, using a previously characterized antibody against 2SC, we show that genetic ablation of FH causes high levels of protein succination. We next hypothesized that immunohistochemistry for 2SC would serve as a metabolic biomarker for the in situ detection of FH-deficient tissues. Robust detection of 2SC was observed in Fh1 (murine FH)-deficient renal cysts and in a retrospective series of HLRCC tumours (n = 16) with established FH mutations. Importantly, 2SC was undetectable in normal tissues (n = 200) and tumour types not associated with HLRCC (n = 1342). In a prospective evaluation of cases referred for genetic testing for HLRCC, the presence of 2SC-modified proteins (2SCP) correctly predicted genetic alterations in FH in every case. In two series of unselected type II papillary renal cancer (PRCC), prospectively analysed by 2SCP staining followed by genetic analysis, the biomarker accurately identified previously unsuspected FH mutations (2/33 and 1/36). The investigation of whether metabolites in other tumour types produce protein modification signature(s) that can be assayed using similar strategies will be of interest in future studies of cancer.
authors.affiliation: Molecular and Population Genetics Laboratory, Wellcome Trust Centre for Human Genetics, Oxford, UK.
authors.forename: Chiara
authors.initials: C
authors.lastname: Bardella
authors.forename: Mona
authors.initials: M
authors.lastname: El-Bahrawy
authors.forename: Norma
authors.initials: N
authors.lastname: Frizzell
authors.forename: Julie
authors.initials: J
authors.lastname: Adam
authors.forename: Nicola
authors.initials: N
authors.lastname: Ternette
authors.forename: Emine
authors.initials: E
authors.lastname: Hatipoglu
authors.forename: Kimberley
authors.initials: K
authors.lastname: Howarth
authors.forename: Linda
authors.initials: L
authors.lastname: O'Flaherty
authors.forename: Ian
authors.initials: I
authors.lastname: Roberts
authors.forename: Gareth
authors.initials: G
authors.lastname: Turner
authors.forename: Jennifer
authors.initials: J
authors.lastname: Taylor
authors.forename: Konstantinos
authors.initials: K
authors.lastname: Giaslakiotis
authors.forename: Valentine M
authors.initials: VM
authors.lastname: Macaulay
authors.forename: Adrian L
authors.initials: AL
authors.lastname: Harris
authors.forename: Ashish
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authors.lastname: Chandra
authors.forename: Heli J
authors.initials: HJ
authors.lastname: Lehtonen
authors.forename: Virpi
authors.initials: V
authors.lastname: Launonen
authors.forename: Lauri A
authors.initials: LA
authors.lastname: Aaltonen
authors.forename: Christopher W
authors.initials: CW
authors.lastname: Pugh
authors.forename: Radu
authors.initials: R
authors.lastname: Mihai
authors.forename: David
authors.initials: D
authors.lastname: Trudgian
authors.forename: Benedikt
authors.initials: B
authors.lastname: Kessler
authors.forename: John W
authors.initials: JW
authors.lastname: Baynes
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Ratcliffe
authors.forename: Ian P
authors.initials: IP
authors.lastname: Tomlinson
authors.forename: Patrick J
authors.initials: PJ
authors.lastname: Pollard
mesh_terms: D000328:Adult; D000368:Aged; D000818:Animals; D014408:Biomarkers, Tumor / Q000172:deficiency / Q000235:genetics / Q000378:metabolism; D002292:Carcinoma, Renal Cell / Q000175:diagnosis* / Q000235:genetics; D004195:Disease Models, Animal; D005260:Female; D005649:Fumarate Hydratase / Q000172:deficiency* / Q000235:genetics / Q000378:metabolism; D020022:Genetic Predisposition to Disease; D018095:Germ-Line Mutation; D006801:Humans; D007680:Kidney Neoplasms / Q000175:diagnosis* / Q000235:genetics; D018231:Leiomyomatosis / Q000175:diagnosis* / Q000235:genetics; D019656:Loss of Heterozygosity; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D008822:Mice, Transgenic; D008875:Middle Aged; D009386:Neoplastic Syndromes, Hereditary / Q000175:diagnosis* / Q000235:genetics; D011446:Prospective Studies; D012680:Sensitivity and Specificity; D019802:Succinic Acid / Q000378:metabolism
chemical_list: D014408:Biomarkers, Tumor; D019802:Succinic Acid; D005649:Fumarate Hydratase