H3.3 K36M Mutation as a Clinical Diagnosis Method of Suspected Chondroblastoma Cases

Abstract

Objective: Whether H3.3 K36M mutation (H3K36M) could be an approach if the diagnosis of chondroblastoma (CB) patients was indistinct and it was suspected to be unclear clinically.

Methods: We reviewed and compared our clinical experiences of CB cases and some suspected cases, which were not diagnosed distinctly, between 2013 to 2019. A total of 15 male and four female cases included in this study were seperated into two groups, CB group and suspected case (SC) group. The CB group included 13 men and 3 women, with an age range from 9 to 54 (mean age, 22 years old). The SC group included two men and one woman, with the age range from 13 to 25 (mean age, 19 years old). In both groups the patients had been followed-up until December 2019 and none of the patients had prior treatment history. We evaluated the clinical complaints, radiological features, and clinical-histological features of the cases and performed an immunohistochemical (IHC) study to detect whether the H3K36M expression of cases was different, consistent with a gene-mutation analysis.

Results: In both groups, the radiologic features of both groups appeared as round low-density shadow with a clear edge, pathologic features showed diffuse proliferation of neoplastic cells with multinuclear giant cells. The radiological tumor size of CB group and SC group showed little difference, which was about 29.0*21.6 mm. Clinical-immunohistochemical features of both groups showed chondroid matrix inside with naïve tumor cells, multinucleated giant cells, and ground substance cells. Most of them showed chondro-related antibody positive (12 cases) but some of them showed S-100 negative (four cases). The clear difference of both groups was the result of H3K36M IHC study and gene analysis. In our cases, the CB group showed diffuse H3K36M positive and the SC group showed negative. The gene mutation analysis revealed that H3K36M-positive CB patients had K36M mutation, which were not found in the SC group. Sanger sequencing showed an A > T substitution at codon 36 of histone H3F3B. No other types of histone H3 mutation was detected in the CB group. Particularly, one of the suspected cases showed a G34W mutation was confirmed to be a giant cell tumor of bone (GCTB).

Conclusions: Our study showed H3K36M immunohistochemistry and gene mutation analysis were specific clinical diagnostic tools to distinguish suspected CB from other giant cell-rich or cartilage matrix-diffuse bone tumors. The clinical-radiological and histomorphological features of patients gave suggestions on whether the H3K36M IHC and gene analysis should be required.

Keywords: Chondroblastoma; Chondromyxoid fibroma; Giant-cell tumor of bone; Histone H3K36 mutation; Immunohistochemistry.

Fig. 1

Clinical‐radiological features of CB cases and suspected cases. (A1‐E1) The X‐ray features of CB cases. (F1) The X‐ray feature of suspected case. (A2‐E2) The CT or MRI features of the corresponding CB cases. (F2) The MRI feature of the corresponding suspected case.

Fig. 2

Clinical‐histological features and H3K36M IHC results of CB cases and suspected case. (A1‐A4) The HE features of CB cases. (B) The HE features of the suspected case. (C1‐C4) The S100 features of the corresponding CB cases. (D) The S100 features of the suspected case. (E1‐E4) The K36M features of the corresponding CB cases. (F) The K36M features of the corresponding the suspected case.

Fig. 3

Clinical‐immunohistochemical features of CB cases and suspected cases. The diagnoses of suspected cases were confirmed by the follow‐up gene analysis. SC: suspected case.

Fig. 4

Gene mutation analysis of CB cases and suspected cases. (A2) Sanger sequencing showed a A > T substitution at codon 36 on histone H3F3B in CB cases. (B2) Sanger sequencing showed a G > T substrtution at codon 34 on histone H3F3A in SC3 (Suspected Case 3).