Journal List > J Korean Soc Endocrinol > v.20(2) > 1063771

Rhee, Woo, Koh, Oh, Kim, Kim, Kim, and Park: Mutational Analysis of Gsα Protein in Fibrous dysplasia of the Bone

Abstract

Background

Fibrous dysplasia of the bone (FD) is a benign fibrous bone lesion which usually involves the long bones of the extremities. FD may be asymptomatic, but often leads to bone deformity and pathological fracture. The disease is caused by a somatic mutation in the Gsα protein, which is responsible for intracellular signal transduction.

Methods

Mutations in the GNAS1 gene, which codes for Gsα protein, was investigated in 34 patients with monostotic and polyostotic FD and McCune-Albright syndrome. DNA was extracted from formalin-fixed, paraffin embedded bone tissues, and exons 8 and 9 of the GNAS1 gene amplified using a polymerase chain reaction (PCR). Subsequently, plasmid cloning and DNA sequencing analysis were performed.

Results

The PCR was successfully performed in 5 patients with monostotic FD. However, the sequencing analysis failed to identify any significant point mutations in exons 8 or 9 of GNAS1. Nevertheless, 3 point mutations were observed in the intron of the GNAS1 gene in 2 samples.

Conclusion

In addition to the previously known somatic mutations of the GNAS1 gene, this study suggests that fibrous dysplasia of the bone might be associated with another point mutations of the GNAS1 gene.

Figures and Tables

Fig. 1
Analyses of gene mutations in patients with fibrous dysplasia. The figures show the intron sequences of the GNAS1 gene. Point mutations (arrowed) were observed in the introns of the 27th and 33rd samples, and these were amplified by using a primer for exon 8. Another point mutation was observed in an intron of the 27th sample, and this was amplified by using a primer for exon 9.
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Table 1
Oligonucleotide Primers Used for PCR Amplification of Exon 8 and Exon 9 in the GNAS1 Gene
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Table 2
Sites of Involved Bone in Patients with Fibrous Dysplasia
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References

1. Lichtenstein L. Polyostotic fibrous dysplasia. Arch Surg. 1938. 36:874–898.
2. Lichtenstein L, Jaffe H. Fibrous dysplasia of a bone: Condition affecting one, several or many bones, graver cases of which may present abnormal pigmentation of skin, premature sexual development, hyperthyroidism or still other extraskeletal abnormalities. Arch Pathol. 1942. 33:777.
3. Fletcher C, Unni K, Mertens F. Pathology and Genetics of Tumours of Soft Tissue and Bone: World Health Organization Classification of Tumours. 2002. Lyon: IARC Press;341–342.
4. Tinschert S, Gerl H, Gewies A, Jung HP, Nurnberg P. McCune-Albright syndrome: Clinical and molecular evidence of mosaicism in an unusual giant patient. Am J Med Genet. 1999. 83:100–108.
5. Weinstein LS, Shenker A, Gejman PV, Merino MJ, Friedman E, Spiegel AM. Activating mutations of the stimulatory G protein in the McCuneAlbright syndrome. N Engl J Med. 1991. 325:1688–1695.
6. Weinstein LS, Yu S, Warner DR, Liu J. Endocrine manifestations of stimulatory G protein alpha-subunit mutations and the role of genomic imprinting. Endocr Rev. 2001. 22:675–705.
7. Coleman DE, Berghuis AM, Lee E, Linder ME, Gilman AG, Sprang SR. Structures of active conformations of Gi alpha 1 and the mechanism of GTP hydrolysis. Science. 1994. 265:1405–1412.
8. Sondek J, Lambright DG, Noel JP, Hamm HE, Sigler PB. GTPase mechanism of Gproteins from the 1.7-A crystal structure of transducin alpha-GDP-AIF-4. Nature. 1994. 372:276–279.
9. Marie PJ, de Pollak C, Chanson P, Lomri A. Increased proliferation of osteoblastic cells expressing the activating Gs alpha mutation in monostotic and polyostotic fibrous dysplasia. Am J Pathol. 1997. 150:1059–1069.
10. Riminucci M, Fisher LW, Shenker A, Spiegel AM, Bianco P, Gehron Robey P. Fibrous dysplasia of bone in the McCune-Albright syndrome: Abnormalities in bone formation. Am J Pathol. 1997. 151:1587–1600.
11. Ringel MD, Schwindinger WF, Levine MA. Clinical implications of genetic defects in G proteins. The molecular basis of McCune-Albright syndrome and Albright hereditary osteodystrophy. Medicine (Baltimore). 1996. 75:171–184.
12. Spiegel AM. Inborn errors of signal transduction: Mutations in G proteins and G protein-coupled receptors as a cause of disease. J Inherit Metab Dis. 1997. 20:113–121.
13. Alman BA, Greel DA, Wolfe HJ. Activating mutations of Gs protein in monostotic fibrous lesions of bone. J Orthop Res. 1996. 14:311–315.
14. Sakamoto A, Oda Y, Iwamoto Y, Tsuneyoshi M. A comparative study of fibrous dysplasia and osteofibrous dysplasia with regard to Gsα mutation at the Arg201 codon: Polymerase chain reaction-restriction fragment length polymorphism analysis of paraffinembedded tissues. J Mol Diagn. 2000. 2:67–72.
15. Bianco P, Riminucci M, Majolagbe A, Kuznetsov SA, Collins MT, Mankani MH, Corsi A, Bone HG, Wientroub S, Spiegel AM, Fisher LW, Robey PG. Mutations of the GNAS1 gene, stromal cell dysfunction, and osteomalacic changes in non-McCune-Albright fibrous dysplasia of bone. J Bone Miner Res. 2000. 15:120–128.
16. Wickham CL, Boyce M, Joyner MV, Sarsfield P, Wilkins BS, Jones DB, Ellard S. Amplification of PCR products in excess of 600 base pairs using DNA extracted from decalcified, paraffin wax embedded bone marrow trephine biopsies. Mol Pathol. 2000. 53:19–23.
17. Provan AB, Hodges E, Smith AG, Smith JL. Use of paraffin wax embedded bone marrow trephine biopsy specimens as a source of archival DNA. J Clin Pathol. 1992. 45:763–765.
18. Wu L, Patten N, Yamashiro CT, Chui B. Extraction and amplification of DNA from formalinfixed, paraffin-embedded tissues. Appl Immunohistochem Mol Morphol. 2002. 10:269–274.
19. Sato Y, Sugie R, Tsuchiya B, Kameya T, Natori M, Mukai K. Comparison of the DNA extraction methods for polymerase chain reaction amplification from formalin-fixed and paraffin-embedded tissues. Diagn Mol Pathol. 2001. 10:265–271.
20. Shi SR, Datar R, Liu C, Wu L, Zhang Z, Cote RJ, Taylor CR. DNA extraction from archival formalinfixed, paraffin-embedded tissues: Heatinduced retrieval in alkaline solution. Histochem Cell Biol. 2004. 122:211–218.
21. Candeliere GA, Roughley PJ, Glorieux FH. Polymerase chain reaction-based technique for the selective enrichment and analysis of mosaic Arg201 mutations in G alpha s from patients with fibrous dysplasia of bone. Bone. 1997. 21:201–206.
22. Alman BA, Naber SP, Terek RM, Jiranek WA, Goldberg MJ, Wolfe HJ. Platelet-derived growth factor in fibrous musculoskeletal disorders: A study of pathologic tissue sections and in vitro primary cell cultures. J Orthop Res. 1995. 13:67–77.
23. Stevens-Simon C, Stewart J, Nakashima II, White M. Exacerbation of fibrous dysplasia associated with an adolescent pregnancy. J Adolesc Health. 1991. 12:403–405.
24. Fraser WD, Walsh CA, Birch MA, Durham B, Dillon JP, McCreavy D, Gallagher JA. Parathyroid hormone-related protein in the aetiology of fibrous dysplasia of bone in the McCune Albright syndrome. Clin Endocrinol (Oxf). 2000. 53:621–628.
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