Diagnostic Utility of Genetic and Immunohistochemical H3-3A Mutation Analysis in Giant Cell Tumour of Bone

• Published in: International journal of molecular sciences 2022-01, Vol.23 (2), p.969
• Main Authors: Wągrodzki, Michał, Tysarowski, Andrzej, Seliga, Katarzyna, Wojnowska, Aneta, Stepaniuk, Maria, Castañeda Wysocka, Patrycja, Makuła, Donata, Pieńkowski, Andrzej, Szostakowski, Bartłomiej, Zub, Renata, Rutkowski, Piotr
• Format: Article
• Language: English
• Subjects: Adolescent; Adult; Aged; Aged, 80 and over; Antibodies; Biopsy; Bone lesions; Bone Neoplasms - diagnosis; Bone Neoplasms - drug therapy; Bone Neoplasms - genetics; Bone Neoplasms - metabolism; Bones; Cell cycle; Child; Cysts; Decalcification; Denosumab - therapeutic use; Diagnosis, Differential; Early Detection of Cancer; Ethylenediaminetetraacetic acids; Female; Gene expression; Genetic analysis; Giant Cell Tumor of Bone - diagnosis; Giant Cell Tumor of Bone - drug therapy; Giant Cell Tumor of Bone - genetics; Giant Cell Tumor of Bone - metabolism; Histone H3; Histones; Histones - genetics; Histones - metabolism; Humans; Male; Middle Aged; Monoclonal antibodies; Mutation; Paraffin Embedding; Paraffins; Patients; Point mutation; Reliability analysis; Sarcoma; Sensitivity; Sensitivity and Specificity; Sequence Analysis, DNA; Tissue Fixation; Tumors; Young Adult
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms23020969

To validate the reliability and implementation of an objective diagnostic method for giant cell tumour of bone (GCTB). gene mutation testing was performed using two different methods, Sanger sequencing and immunohistochemical (IHC) assays. A total of 214 patients, including 120 with GCTB and 94 with other giant cell-rich bone lesions, participated in the study. Sanger sequencing and IHC with anti-histone H3.3 G34W and G34V antibodies were performed on formalin-fixed, paraffin-embedded tissues, which were previously decalcified in EDTA if needed. The sensitivity and specificity of the molecular method was 100% (95% CI: 96.97-100%) and 100% (95% CI: 96.15-100%), respectively. The sensitivity and specificity of IHC was 94.32% (95% CI: 87.24-98.13%) and 100% (95% CI: 93.94-100.0%), respectively. P.G35 mutations were discovered in 2/9 (22.2%) secondary malignant GCTBs and 9/13 (69.2%) GCTB after denosumab treatment. We confirmed in a large series of patients that evaluation of mutational status using direct sequencing is a reliable tool for diagnosing GCTB, and it should be incorporated into the diagnostic algorithm. Additionally, we discovered IHC can be used as a screening tool. Proper tissue processing and decalcification are necessary. The presence of the mutation did not exclude malignant GCTB. Denosumab did not eradicate the neoplastic cell population of GCTB.