Volume 225, Issue 1 p. 4-11
Original Paper

Aberrant succination of proteins in fumarate hydratase-deficient mice and HLRCC patients is a robust biomarker of mutation status

Chiara Bardella

Chiara Bardella

Molecular and Population Genetics Laboratory, Wellcome Trust Centre for Human Genetics, Oxford, UK

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Mona El-Bahrawy

Mona El-Bahrawy

Department of Histopathology, Imperial College, Hammersmith Hospital, London, UK

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Norma Frizzell

Norma Frizzell

Department of Exercise Science, School of Public Health, University of South Carolina, Columbia, SC, USA

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Julie Adam

Julie Adam

Henry Wellcome Building for Molecular Physiology, University of Oxford, UK

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Nicola Ternette

Nicola Ternette

Henry Wellcome Building for Molecular Physiology, University of Oxford, UK

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Emine Hatipoglu

Emine Hatipoglu

Henry Wellcome Building for Molecular Physiology, University of Oxford, UK

These authors contributed equally to this work.

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Kimberley Howarth

Kimberley Howarth

Molecular and Population Genetics Laboratory, Wellcome Trust Centre for Human Genetics, Oxford, UK

These authors contributed equally to this work.

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Linda O'Flaherty

Linda O'Flaherty

Henry Wellcome Building for Molecular Physiology, University of Oxford, UK

These authors contributed equally to this work.

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Ian Roberts

Ian Roberts

Department of Cellular Pathology, John Radcliffe Hospital, Oxford, UK

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Gareth Turner

Gareth Turner

Department of Cellular Pathology, John Radcliffe Hospital, Oxford, UK

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Jennifer Taylor

Jennifer Taylor

Oxford Biomedical Research Centre, Wellcome Trust Centre for Human Genetics, Oxford, UK

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Konstantinos Giaslakiotis

Konstantinos Giaslakiotis

Department of Cellular Pathology, John Radcliffe Hospital, Oxford, UK

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Valentine M Macaulay

Valentine M Macaulay

Weatherall Institute of Molecular Medicine, University of Oxford, UK

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Adrian L Harris

Adrian L Harris

Weatherall Institute of Molecular Medicine, University of Oxford, UK

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Ashish Chandra

Ashish Chandra

Cellular Pathology, St Thomas' Hospital, London, UK

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Heli J Lehtonen

Heli J Lehtonen

Department of Medical Genetics, Biomedicum Helsinki, University of Helsinki, Finland

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Virpi Launonen

Virpi Launonen

Department of Medical Genetics, Biomedicum Helsinki, University of Helsinki, Finland

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Lauri A Aaltonen

Lauri A Aaltonen

Department of Medical Genetics, Biomedicum Helsinki, University of Helsinki, Finland

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Christopher W Pugh

Christopher W Pugh

Henry Wellcome Building for Molecular Physiology, University of Oxford, UK

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Radu Mihai

Radu Mihai

Department of Endocrine Surgery, John Radcliffe Hospital, Oxford, UK

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David Trudgian

David Trudgian

Henry Wellcome Building for Molecular Physiology, University of Oxford, UK

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Benedikt Kessler

Benedikt Kessler

Henry Wellcome Building for Molecular Physiology, University of Oxford, UK

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John W Baynes

John W Baynes

Department of Exercise Science, School of Public Health, University of South Carolina, Columbia, SC, USA

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Peter J Ratcliffe

Peter J Ratcliffe

Henry Wellcome Building for Molecular Physiology, University of Oxford, UK

These authors contributed equally to this work.

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Ian P Tomlinson

Ian P Tomlinson

Molecular and Population Genetics Laboratory, Wellcome Trust Centre for Human Genetics, Oxford, UK

Oxford Biomedical Research Centre, Wellcome Trust Centre for Human Genetics, Oxford, UK

These authors contributed equally to this work.

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Patrick J Pollard

Corresponding Author

Patrick J Pollard

Henry Wellcome Building for Molecular Physiology, University of Oxford, UK

Henry Wellcome Building for Molecular Physiology, University of Oxford, Oxford, UK.Search for more papers by this author
First published: 10 May 2011
Citations: 216

Conflict of interest statement. PP, NF, JWB and IPT have filed for a patent [No. USC-268-P(849)] covering immunohistochemical screening for the determination of FH mutations.

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. Copyright © 2011 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.