Methylation Analysis Reveals Epigenetic Congruence Between Bone Sarcomas With H3-3A Mutations and Malignant Giant Cell Tumors of Bone

Hotspot mutations in H3-3A gene are key drivers in giant cell tumor of bone (GCTB). Rare primary bone sarcomas also harbor this mutation, but their clinicopathologic characteristics and molecular profiles, as well as their relationship to conventional and malignant GCTB (MGCTB) and high-grade conventional osteosarcoma (HGOS), are largely undefined. Herein, we present a series of 10 H3-3A mutated bone sarcomas (BSH3-3A) with a comparative clinicopathologic, mutational, and epigenetic analysis with conventional GCTB, MGCTB, and HGOS. BSH3-3A comprised of 6 high-grade osteosarcomas, 4 undifferentiated pleomorphic sarcoma of bone and occurred in 7 women and 3 men with a mean age of 46 years (28-74 years). The tumors involved femur (n = 4), talus (n = 2), spine (n = 2), pelvis (n = 1), and 1 unknown site. Epiphysis involvement was noted in 2 femoral tumors. In majority of the cases, BSH3-3A showed cellular proliferation of epithelioid and/or spindle cells, hyperchromatic nuclei, and conspicuous pleomorphism with or without osteoid production. One case exhibited both low- and high-grade osteosarcoma components. The mutational profile of BSH3-3A was different than that of conventional HGOS with significantly less frequent TP53 mutations. The genomic index, which reflects the degree of genomic complexity, was also significantly lower in BSH3-3A compared with HGOS, yet higher than GCTB. DNA methylation analysis revealed that most BSH3-3A and MGCTB cases form a distinct cluster, positioned near but separate from GCTB, and clearly separated from HGOS. Differential methylation analysis revealed that BSH3-3A exhibited the highest degree of similarity to MGCTB in comparison to HGOS and GCTB. Survival analysis showed that outcomes for BSH3-3A do not differ significantly from those observed in HGOS or MGCTB. Finally, BSH3-3A tumors, although radiologically and histologically identical to high-grade bone sarcomas lacking H3-3A mutations, display epigenetic features similar to MGCTB and have a significantly less complex genomic profile than HGOS, despite comparable clinical outcomes.