Mutation Analysis of H3F3A and H3F3B as a Diagnostic Tool for Giant Cell Tumor of Bone and Chondroblastoma

Abstract

Specific H3F3A driver mutations and IDH2 mutations were recently described in giant cell tumor of bone (GCTB) and H3F3B driver mutations in chondroblastoma; these may be helpful as a diagnostic tool for giant cell-containing tumors of the bone. Using Sanger sequencing, we determined the frequency of H3F3A, H3F3B, IDH1, and IDH2 mutations in GCTBs (n=60), chondroblastomas (n=12), and other giant cell-containing tumors (n=24), including aneurysmal bone cyst, chondromyxoid fibroma, and telangiectatic osteosarcoma. To find an easy applicable marker for H3F3A mutation status, H3K36 trimethylation and ATRX expression were correlated with H3F3A mutations. In total, 69% of all GCTBs harbored an H3F3A (G34W/V) mutation compared with 0% of all other giant cell-containing tumors (P<0.001), whereas 70% of chondroblastomas showed an H3F3B (K36M) mutation compared with 0% of other giant cell-containing tumors (P<0.001). Diffuse H3K36 trimethylation positivity was more often seen in mutated H3F3A GCTBs compared with other giant cell-containing tumors (P=0.005). ATRX protein expression was not correlated with H3F3A mutation status. Hotspot mutations in IDH1 or IDH2 were absent. Our results show that H3F3A and H3F3B mutation analysis appears to be a highly specific, although less sensitive, diagnostic tool for the distinction of GCTB and chondroblastoma from other giant cell-containing tumors. Although H3K36 trimethylation and ATRX immunohistochemistry cannot be used as surrogate markers for H3F3A mutation status, mutations in H3F3A are associated with increased H3K36 trimethylation, suggesting that methylation at this residue may play a role in the etiology of the disease.

Development of high-grade osteosarcoma in a patient with recurrent giant cell tumor of the ischium while receiving treatment with denosumab

“…The diagnosis of giant cell-containing lesions of bone is challenging (15). Cleven et al (15) recently reported that H3F3A mutation analysis appears to be a highly specific, although less sensitive, diagnostic tool for the distinction of GCTB from other giant cellcontaining tumors.…”

“…Cleven et al (15) recently reported that H3F3A mutation analysis appears to be a highly specific, although less sensitive, diagnostic tool for the distinction of GCTB from other giant cellcontaining tumors. Agaram et al (9) reported that fluorescence in situ hybridization for USP6 break-apart is a useful ancillary tool in the diagnosis of primary aneurysmal bone cysts and can help to distinguish them from giant cell reparative granulomas and other morphologically similar lesions.…”

CORR ® Tumor Board: What are the Functional Results, Complications, and Outcomes of Using a Custom Unipolar Wrist Hemiarthroplasty for Treatment of Grade III Giant Cell Tumors of the Distal Radius?

“…[2,4]. This finding has helped to confirm cases with unusual histologic features or metaphyseal location as bona fide giant cell tumors, expanding the clinicopathologic spectrum of the disease [1,4].…”

“…[2,4]. This finding has helped to confirm cases with unusual histologic features or metaphyseal location as bona fide giant cell tumors, expanding the clinicopathologic spectrum of the disease [1,4]. H3F3A encodes a replicationindependent histone (H3.3), known to have an epigenetic function regulating other genes; however, the exact mechanistic role of H3F3A mutation in the development of giant cell tumor is unknown.…”

Role of Next-Generation Sequencing as a Diagnostic Tool for the Evaluation of Bone and Soft-Tissue Tumors

“…At least in our experience, IHC is of limited help in difficult cases. However, recent advances in molecular pathology using NGS demonstrated K36M mutations in the H3F3B gene in 95% of chondroblastomas and H3F3A mutations in 92% of giant cell tumors of the bones [12,13] . Both genes encode for histone H3.3 [14] .…”