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Mutations in SH3BP2, the cherubism gene, were not detected in central or peripheral giant cell tumours of the jaw
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British Journal of Oral and Maxillofacial Surgery 46 (2008) 229–230
Short communication
Mutations in SH3BP2, the cherubism gene, were not detected in central or peripheral giant cell tumours of the jaw Bernadine D. Idowu a,b , Garreth Thomas c,d , Richard Frow c , Timothy C. Diss b , Adrienne M. Flanagan a,b,c,d,∗ a
Institute of Orthopaedics and Musculoskeletal Science, University College London, Stanmore, Middlesex, UK Histopathology, Royal National Orthopaedic Hospital NHS Trust, Stanmore, UK c Department of Histopathology, University College Hospital, London, UK d Tumour Biology Laboratory, Cancer Research UK Clinical Centre, Barts and the London Queen Mary’s University, UK b
Accepted 15 April 2007 Available online 4 June 2007
Abstract Giant cell granulomas of the jaw (GCGJ) are non-familial, generally unilateral osteoclast-rich lesions that are histopathologically indistinguishable from cherubism. Cherubism is an autosomal dominant disease that is characterised by bilateral radiolucencies of the jaw, and caused by mutations that occur in SH3BP2 exon 10. The aim of the study was to screen lesional GCGJ tissue for SH3BP2 mutations. Lesional mononuclear stromal or spindle cells were microdissected from paraffin-embedded tissue from GCGJ, and DNA was then extracted and sequenced for SH3BP2 mutations associated with cherubism. No mutations were detected in 26 GCGJ (15 central, 11 peripheral), which indicated that people with GCGJ do not harbour cherubism-related germline SH3BP2 mutations, and that GCGJ do not harbour somatic SH3BP2 mutations. This suggests that cherubism and GCGJ arise on a different genetic background, and therefore detection of SH3BP2 mutations can be a useful means of distinguishing between them. © 2007 The British Association of Oral and Maxillofacial Surgeons. Published by Elsevier Ltd. All rights reserved. Keywords: Cherubism; Giant cell granuloma of jaw; Mutation; SH3BP2
Introduction Giant cell granuloma of the jaw (GCGJ) is classified as either peripheral or central, both of which are non-familial. Central GCGJ is an osteolytic lesion of the mandible or maxilla and behaves in a variably aggressive manner. The peripheral variant occurs on the gingiva or alveolar ridge more commonly than in the mandible. Cherubism (OMIM 118400) is an autosomal dominant disease that is present in childhood, and is characterised by bilateral radiolucencies of the mandible and sometimes the maxilla. Ueki et al. showed that germline mutations in SH3BP2 (ENST0000310699) are implicated in its pathogenesis.1 ∗
Corresponding author. Tel.: +44 20 8909 5354. E-mail address: [email protected] (A.M. Flanagan).
GCGJ and cherubism cannot be distinguished microscopically,2 and as there are occasional reports of unilateral cherubism,3 and of cases of multiple central GCGJ that occur in the absence of hyperparathyroidism,4 it is of interest to know if patients with cherubism and GCGJ share the same genetic abnormality either at a germline or somatic level. Germline mutations in SH3BP2 have not been detected in blood samples from four patients with central GCGJ.2 The aim of this study was to investigate whether somatic SH3BP2 mutations occur in GCGJ.
Materials and methods All GCGJ included were unilateral lesions: hyperparathyroidism had been excluded. Twenty six GCGJ, 11 peripheral (4 men, 7 women), and 15 central (5 men, 10 women) were
0266-4356/$ – see front matter © 2007 The British Association of Oral and Maxillofacial Surgeons. Published by Elsevier Ltd. All rights reserved.
doi:10.1016/j.bjoms.2007.04.014
230
B.D. Idowu et al. / British Journal of Oral and Maxillofacial Surgery 46 (2008) 229–230
analysed. The age at presentation of the peripheral lesions ranged between 26 and 73 years (mean 45), and for those with central lesions between 11 and 43 years (mean 28). Nine of the peripheral lesions were on the mandible and 2 on the maxilla: 7 and 8 of the central lesions were at these sites, respectively. The study complied with the standards of the local ethics committee. Five sections 5 m thick of each tumour were stained with haematoxylin, and their mononuclear spindle cells microdissected using a 14G needle under light microscopy. Roughly 80% of the DNA analysed was derived from this group and contamination with other tissue (including the osteoclast-like giant cells that are a reactive population and would not harbour a mutation)5 was kept to a minimum. DNA was extracted using Proteinase K (Sigma, Poole, Dorset, UK). Primers were designed to flank the mutation-rich region in SH2BP3 exon 10. The forward primer was sited in the intronic region of exon 9 (forward primer – 5 CCG CCC CGT GTC TGA CAG TGA3 , reverse primer – 5 AGT GTC AGC CTG TGA GGG TTG3 ). The primers designed for the original polymerase chain reaction were used for the sequencing cycling reaction. Each case was sequenced in both directions using the DTCS Quick Start kit (Beckman Coulter UK Ltd., Bucks, UK) and analysed on a CEQTM 8000 DNA Genetic Analysis System (Beckman Coulter).
Results The expected 130 bp product of polymerase chain reaction was generated from all genomic DNA samples extracted from the mononuclear spindle cell component of the tumours. However, direct sequencing failed to show any of the previously reported missense SH3BP2 mutations. DNA extracted from blood, and paraffin-embedded tissue from the jaw lesion of a patient with cherubism were used as controls, and showed a PRO418ARG mutation in both samples.
Discussion To our knowledge this is the first report that illustrates the failure to detect cherubism-associated mutations in GCGJ, which implies that neither germline nor somatic SH3BP2 cherubism-associated mutations are implicated in the pathogenesis of GCGJ, and that these morphologically-similar lesions are genetically distinct. In addition to SH3BP2, germline mutations in NF1 and PTPN11 are also implicated in the pathogenesis of GCGJ,
and are central to the development of neurofibromatosis-type 1 and Noonan syndrome, respectively; these syndromes may present with a cherubism phenotype, or with single or multiple GCGJ.6–8 Although these syndromes may be familial, this is not always the case, and therefore they should be included in the differential diagnosis in patients who present with osteoclast-rich lesions of the jaw. Recent studies in mice predicted that patients with cherubism can be treated successfully with anti-tumour necrosis factor-␣ (TNF-␣) agents.9 To investigate this it will be important to distinguish between cherubism and GCGJ, because if the mechanism by which these lesions develop are different, GCGJ may not respond to agents directed against TNF-␣. Acknowledgements We thank Chris Yeoman for his expert technical assistance, and Research and Development Unit, The Royal National Orthopaedic Hospital, Stanmore, UK and The Mason Medical Research Foundation for financial support.
References 1. Ueki Y, Tiziani V, Santanna C, Fukai N, Maulik C, Garfinkle J, et al. Mutations in the gene encoding c-Abl-binding protein SH3BP2 cause cherubism. Nature Genet 2001;28:125–6. 2. de Lange J, van Maarle MC, van den Akker HP, Redeker EJ., DNA analysis of the SH3BP2 gene in patients with aggressive central giant cell granuloma. Br J Oral Maxillofac Surg; Epub ahead of print, in press. 3. Mangion J, Edkins S, Goss AN, Stratton MR, Flanagan AM. Familial craniofacial fibrous dysplasia: absence of linkage to GNAS1 and the gene for cherubism. J Med Genet 2000;37:E37. 4. de Lange J, van den Akker HP. Clinical and radiological features of central giant-cell lesions of the jaw. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2005;99:464–70. 5. Flanagan AM, Tinkler SM, Horton MA, Williams DM, Chambers TJ. The multinucleate cells in giant cell granulomas of the jaw are osteoclasts. Cancer 1988;62:1139–45. 6. Tartaglia M, Kalidas K, Shaw A, Song X, Musat DL, van der Burgt I, et al. PTPN11 mutations in Noonan syndrome: molecular spectrum, genotypephenotype correlation, and phenotypic heterogenetity. Am J Hum Genet 2002;70:1555–63. 7. van Capelle CI, Hogeman PH, van der Sijs-Bos CJ, Heggelman BG, Idowu B, Slootweg PJ, et al., Neurofibromatosis presenting with a cherubism phenotype. Eur J Pediatr; Epub ahead of print, in press. 8. Jafarov T, Ferimazova N, Reichenberger E. Noonan-like syndrome mutations in PTPN11 in patients diagnosed with cherubism. Clin Genet 2005;68:190–1. 9. Ueki Y, Lin CY, Senoo M, Ebihara T, Agata N, Onji M, et al. Increased myeloid cell responses to M-CSF and RANKL cause bone loss and inflammation in SH3BP2 “cherubism” mice. Cell 2007;128:71–83.
British Journal of Oral and Maxillofacial Surgery 46 (2008) 229–230
Short communication
Mutations in SH3BP2, the cherubism gene, were not detected in central or peripheral giant cell tumours of the jaw Bernadine D. Idowu a,b , Garreth Thomas c,d , Richard Frow c , Timothy C. Diss b , Adrienne M. Flanagan a,b,c,d,∗ a
Institute of Orthopaedics and Musculoskeletal Science, University College London, Stanmore, Middlesex, UK Histopathology, Royal National Orthopaedic Hospital NHS Trust, Stanmore, UK c Department of Histopathology, University College Hospital, London, UK d Tumour Biology Laboratory, Cancer Research UK Clinical Centre, Barts and the London Queen Mary’s University, UK b
Accepted 15 April 2007 Available online 4 June 2007
Abstract Giant cell granulomas of the jaw (GCGJ) are non-familial, generally unilateral osteoclast-rich lesions that are histopathologically indistinguishable from cherubism. Cherubism is an autosomal dominant disease that is characterised by bilateral radiolucencies of the jaw, and caused by mutations that occur in SH3BP2 exon 10. The aim of the study was to screen lesional GCGJ tissue for SH3BP2 mutations. Lesional mononuclear stromal or spindle cells were microdissected from paraffin-embedded tissue from GCGJ, and DNA was then extracted and sequenced for SH3BP2 mutations associated with cherubism. No mutations were detected in 26 GCGJ (15 central, 11 peripheral), which indicated that people with GCGJ do not harbour cherubism-related germline SH3BP2 mutations, and that GCGJ do not harbour somatic SH3BP2 mutations. This suggests that cherubism and GCGJ arise on a different genetic background, and therefore detection of SH3BP2 mutations can be a useful means of distinguishing between them. © 2007 The British Association of Oral and Maxillofacial Surgeons. Published by Elsevier Ltd. All rights reserved. Keywords: Cherubism; Giant cell granuloma of jaw; Mutation; SH3BP2
Introduction Giant cell granuloma of the jaw (GCGJ) is classified as either peripheral or central, both of which are non-familial. Central GCGJ is an osteolytic lesion of the mandible or maxilla and behaves in a variably aggressive manner. The peripheral variant occurs on the gingiva or alveolar ridge more commonly than in the mandible. Cherubism (OMIM 118400) is an autosomal dominant disease that is present in childhood, and is characterised by bilateral radiolucencies of the mandible and sometimes the maxilla. Ueki et al. showed that germline mutations in SH3BP2 (ENST0000310699) are implicated in its pathogenesis.1 ∗
Corresponding author. Tel.: +44 20 8909 5354. E-mail address: [email protected] (A.M. Flanagan).
GCGJ and cherubism cannot be distinguished microscopically,2 and as there are occasional reports of unilateral cherubism,3 and of cases of multiple central GCGJ that occur in the absence of hyperparathyroidism,4 it is of interest to know if patients with cherubism and GCGJ share the same genetic abnormality either at a germline or somatic level. Germline mutations in SH3BP2 have not been detected in blood samples from four patients with central GCGJ.2 The aim of this study was to investigate whether somatic SH3BP2 mutations occur in GCGJ.
Materials and methods All GCGJ included were unilateral lesions: hyperparathyroidism had been excluded. Twenty six GCGJ, 11 peripheral (4 men, 7 women), and 15 central (5 men, 10 women) were
0266-4356/$ – see front matter © 2007 The British Association of Oral and Maxillofacial Surgeons. Published by Elsevier Ltd. All rights reserved.
doi:10.1016/j.bjoms.2007.04.014
230
B.D. Idowu et al. / British Journal of Oral and Maxillofacial Surgery 46 (2008) 229–230
analysed. The age at presentation of the peripheral lesions ranged between 26 and 73 years (mean 45), and for those with central lesions between 11 and 43 years (mean 28). Nine of the peripheral lesions were on the mandible and 2 on the maxilla: 7 and 8 of the central lesions were at these sites, respectively. The study complied with the standards of the local ethics committee. Five sections 5 m thick of each tumour were stained with haematoxylin, and their mononuclear spindle cells microdissected using a 14G needle under light microscopy. Roughly 80% of the DNA analysed was derived from this group and contamination with other tissue (including the osteoclast-like giant cells that are a reactive population and would not harbour a mutation)5 was kept to a minimum. DNA was extracted using Proteinase K (Sigma, Poole, Dorset, UK). Primers were designed to flank the mutation-rich region in SH2BP3 exon 10. The forward primer was sited in the intronic region of exon 9 (forward primer – 5 CCG CCC CGT GTC TGA CAG TGA3 , reverse primer – 5 AGT GTC AGC CTG TGA GGG TTG3 ). The primers designed for the original polymerase chain reaction were used for the sequencing cycling reaction. Each case was sequenced in both directions using the DTCS Quick Start kit (Beckman Coulter UK Ltd., Bucks, UK) and analysed on a CEQTM 8000 DNA Genetic Analysis System (Beckman Coulter).
Results The expected 130 bp product of polymerase chain reaction was generated from all genomic DNA samples extracted from the mononuclear spindle cell component of the tumours. However, direct sequencing failed to show any of the previously reported missense SH3BP2 mutations. DNA extracted from blood, and paraffin-embedded tissue from the jaw lesion of a patient with cherubism were used as controls, and showed a PRO418ARG mutation in both samples.
Discussion To our knowledge this is the first report that illustrates the failure to detect cherubism-associated mutations in GCGJ, which implies that neither germline nor somatic SH3BP2 cherubism-associated mutations are implicated in the pathogenesis of GCGJ, and that these morphologically-similar lesions are genetically distinct. In addition to SH3BP2, germline mutations in NF1 and PTPN11 are also implicated in the pathogenesis of GCGJ,
and are central to the development of neurofibromatosis-type 1 and Noonan syndrome, respectively; these syndromes may present with a cherubism phenotype, or with single or multiple GCGJ.6–8 Although these syndromes may be familial, this is not always the case, and therefore they should be included in the differential diagnosis in patients who present with osteoclast-rich lesions of the jaw. Recent studies in mice predicted that patients with cherubism can be treated successfully with anti-tumour necrosis factor-␣ (TNF-␣) agents.9 To investigate this it will be important to distinguish between cherubism and GCGJ, because if the mechanism by which these lesions develop are different, GCGJ may not respond to agents directed against TNF-␣. Acknowledgements We thank Chris Yeoman for his expert technical assistance, and Research and Development Unit, The Royal National Orthopaedic Hospital, Stanmore, UK and The Mason Medical Research Foundation for financial support.
References 1. Ueki Y, Tiziani V, Santanna C, Fukai N, Maulik C, Garfinkle J, et al. Mutations in the gene encoding c-Abl-binding protein SH3BP2 cause cherubism. Nature Genet 2001;28:125–6. 2. de Lange J, van Maarle MC, van den Akker HP, Redeker EJ., DNA analysis of the SH3BP2 gene in patients with aggressive central giant cell granuloma. Br J Oral Maxillofac Surg; Epub ahead of print, in press. 3. Mangion J, Edkins S, Goss AN, Stratton MR, Flanagan AM. Familial craniofacial fibrous dysplasia: absence of linkage to GNAS1 and the gene for cherubism. J Med Genet 2000;37:E37. 4. de Lange J, van den Akker HP. Clinical and radiological features of central giant-cell lesions of the jaw. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2005;99:464–70. 5. Flanagan AM, Tinkler SM, Horton MA, Williams DM, Chambers TJ. The multinucleate cells in giant cell granulomas of the jaw are osteoclasts. Cancer 1988;62:1139–45. 6. Tartaglia M, Kalidas K, Shaw A, Song X, Musat DL, van der Burgt I, et al. PTPN11 mutations in Noonan syndrome: molecular spectrum, genotypephenotype correlation, and phenotypic heterogenetity. Am J Hum Genet 2002;70:1555–63. 7. van Capelle CI, Hogeman PH, van der Sijs-Bos CJ, Heggelman BG, Idowu B, Slootweg PJ, et al., Neurofibromatosis presenting with a cherubism phenotype. Eur J Pediatr; Epub ahead of print, in press. 8. Jafarov T, Ferimazova N, Reichenberger E. Noonan-like syndrome mutations in PTPN11 in patients diagnosed with cherubism. Clin Genet 2005;68:190–1. 9. Ueki Y, Lin CY, Senoo M, Ebihara T, Agata N, Onji M, et al. Increased myeloid cell responses to M-CSF and RANKL cause bone loss and inflammation in SH3BP2 “cherubism” mice. Cell 2007;128:71–83.