Frequency of GNAS R201H substitution mutation in polyostotic fibrous dysplasia: Pyrosequencing analysis in tissue samples with or without decalcification

• Published in: Scientific reports 2017-06, Vol.7 (1), p.2836-7, Article 2836
• Main Authors: Shin, Su-Jin, Lee, Seok Joo, Kim, Sang Kyum
• Format: Article
• Language: English
• Subjects: 38/23; 38/39; 38/77; 692/4017; 692/699/67/1344; Adolescent; Adult; Aged; Aged, 80 and over; Amino Acid Substitution; Biopsy; Child; Child, Preschool; Chromogranins - genetics; Decalcification; Deoxyribonucleic acid; DNA; DNA Mutational Analysis; Female; Fibrous dysplasia; Fibrous Dysplasia, Polyostotic - genetics; Fibrous Dysplasia, Polyostotic - pathology; Gene Frequency; GTP-Binding Protein alpha Subunits, Gs - genetics; Guanine; Guanine nucleotide-binding protein; High-Throughput Nucleotide Sequencing; Humanities and Social Sciences; Humans; Infant; Male; Middle Aged; multidisciplinary; Mutation; Paraffin; Pathogenesis; Science; Science (multidisciplinary); Sequence Analysis, DNA; Young Adult
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-017-03093-1

Guanine nucleotide-binding protein/α-subunit ( GNAS ) mutations are involved in fibrous dysplasia (FD) pathogenesis. Here, we analyzed GNAS mutations in FD which were performed by pyrosequencing DNA isolated from formalin-fixed paraffin-embedded (FFPE) tissue. The mutation detection rate was determined in FD specimens with and without decalcification. GNAS mutation was identified in 28 cases out of 87 FDs (32.18%) [p.R201C ( N  = 14) and p.R201H ( N  = 14)]. GNAS mutation was more likely to occur in polyostotic FD (7/28, 25.0%); FD without GNAS mutation was mostly monostotic form (56/59, 94.9%, P  = 0.011). The G > A (R201H) mutation was more frequent in polyostotic FD (6/14 patients, 42.9%) than the C > T (R201C) mutation (1/14, 7.1%) ( P  = 0.077). We divided the FD cases into two subgroups: tissue specimens that were not decalcified ( N  = 35, 40.2%), and tissue specimens that were decalcified ( N  = 52, 59.8%). GNAS mutation was more frequently identified in FD specimens that were not subjected to decalcification (23/35, 65.7%) than in FD specimens that were decalcified (5/52, 9.6%) ( P  = 0.001). In conclusion, mutation analysis of GNAS by pyrosequencing has diagnostic value in FFPE tissue of patients with FD, especially in specimens that were not decalcified. The R201H substitution mutation of GNAS may be involved in the pathogenesis of polyostotic FD.