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. 2014 Nov;128(5):743-53.
doi: 10.1007/s00401-014-1338-3. Epub 2014 Sep 9.

A Sensitive and Specific Histopathologic Prognostic Marker for H3F3A K27M Mutant Pediatric Glioblastomas

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Free PMC article

A Sensitive and Specific Histopathologic Prognostic Marker for H3F3A K27M Mutant Pediatric Glioblastomas

Sriram Venneti et al. Acta Neuropathol. .
Free PMC article

Abstract

Pediatric glioblastomas (GBM) are highly aggressive and lethal tumors. Recent sequencing studies have shown that ~30 % of pediatric GBM and ~80 % of diffuse intrinsic pontine gliomas show K27M mutations in the H3F3A gene, a variant encoding histone H3.3. H3F3A K27M mutations lead to global reduction in H3K27me3. Our goal was to develop biomarkers for the histopathologic detection of these tumors. Therefore, we evaluated the utility of measuring H3K27me3 global reduction as a histopathologic and prognostic biomarker and tested an antibody directed specifically against the H3.3 K27M mutation in 290 samples. The study cohort included 203 pediatric (including 38 pediatric high-grade astrocytomas) and 38 adult brain tumors of various subtypes and grades and 49 non-neoplastic reactive brain tissues. Detection of H3.3 K27M by immunohistochemistry showed 100 % sensitivity and specificity and was superior to global reduction in H3K27me3 as a biomarker in diagnosing H3F3A K27M mutations. Moreover, cases that stained positive for H3.3 K27M showed a significantly poor prognosis compared to corresponding negative tumors. These results suggest that immunohistochemical detection of H3.3 K27M is a sensitive and specific surrogate for the H3F3A K27M mutation and defines a prognostically poor subset of pediatric GBM.

Figures

Fig. 1
Fig. 1
Comparison of immunostaining for H3K27me3 and H3.3 K27M in wild-type and H3F3A K27M mutant pediatric GBM. a, b Representative images from a H3F3A K27M wild-type tumor, H3K27me3 (40×, a) and H3.3 K27M (40×, b). cf. Representative images from two different H3F3A K27M mutant tumors, H3K27me3 (40×, c and 60×, e) and H3.3 K27M (40×, d and 60×, f). Arrowheads indicate endothelial cells in blood vessels
Fig. 2
Fig. 2
Quantification of H3K27me3 and H3.3 K27M immunostaining in adult and pediatric brain tumors and survival analyses in pediatric high-grade gliomas. a, c Quantification of H3K27me3 (a) and H3.3 K27M (c) IHC in pediatric brain tumors. ***p < 0.0001. Pilo pilocytic astrocytoma, SEGA subependymal giant cell astrocytoma, Astro grade II astrocytoma, OD oligodendroglioma, DNET dysembryoplastic neuroepithelial tumor, GG ganglioglioma, NC neurocytoma, MEN meningioma, CP craniopharyngioma, AT/RT atypical teratoid/rhabdoid tumor, CPP choroid plexus papilloma, MB medulloblastoma, K27M H3F3A K27M mutant GBMs, WT GBMs without H3F3A K27M mutations. b Quantification of H3K27me3 (left Y-axis) and H3.3 K27M (right Y-axis) in adult brain tumors. Pilo pilocytic astrocytoma, DA diffuse astrocytoma, GG ganglioglioma, AA anaplastic astrocytomas, GBM glioblastoma, OD oligodendroglioma, AO anaplastic oligodendrogliomas. d Kaplan–Meier survival analyses between H3.3 K27M immunopositive (red line) and H3.3 K27M immunonegative (black line) pediatric GBMs (see also Table 2)
Fig. 3
Fig. 3
Comparison of H3K27me3 and H3.3 K27M in CNS embryonal tumors. a, b Representative images of H3K27me3 (40×, a) and H3.3 K27M (40×, b) in medulloblastoma. c, d Representative images of H3K27me3 (40×, c) and H3.3 K27M (40×, d) in an AT/RT case with high H3K27me3. e , f Representative images of H3K27me3 (40×, e) and H3.3 K27M (40×, f) in subset of AT/RT with low H3K27me3
Fig. 4
Fig. 4
Comparison of H3K27me3 and H3.3 K27M in pediatric brain tumors. a, b Representative images of H3K27me3 (40×, a) and H3.3 K27M (40×, b) in choroid plexus papilloma (CPC). c, d Representative images of H3K27me3 (40×, c) and H3.3 K27M (40×, d) in meningioma. e, f Representative images of H3K27me3 (40×, e) and H3.3 K27M (40×, f) in craniopharyngioma. g, h Representative images of H3K27me3 (40×, g) and H3.3 K27M (40×, h) in neurocytoma
Fig. 5
Fig. 5
Comparison of H3K27me3 and H3.3 K27M in adult brain tumors. a, b Representative images of H3K27me3 (40×, a) and H3.3 K27M (40×, b) in diffuse astrocytoma. c, d Representative images of H3K27me3 (40×, c) and H3.3 K27M (40×, d) in anaplastic oligodendroglioma e and f. Representative images of H3K27me3 (40×, e) and H3.3 K27M (40×, f) in anaplastic astrocytoma. g, h Representative images of H3K27me3 (40×, g) and H3.3 K27M (40×, h) in adult GBM
Fig. 6
Fig. 6
Comparison of H3K27me3 and H3.3 K27M in non-neoplastic brain tissues. a, b Representative images of H3K27me3 (5×, a, inset 40×) and H3.3 K27M (5×, b, inset 40×) in focal cortical dysplasia (FCD). c, d Representative images of H3K27me3 (10×, c, inset 40×) and H3.3 K27M (10×, d, inset 40×) in hippocampal (Hip) sclerosis. e, f Representative images of H3K27me3 (5×, e, inset 40×) and H3.3 K27M (5×, f, inset 40×) in hemorrhage. g, h Representative images of H3K27me3 (5×, g, inset 40×) and H3.3 K27M (5×, h, inset 40×) in abscess, inset shows abscess wall. i, j Representative images of H3K27me3 (5×, i, inset 40×) and H3.3 K27M (5×, j, inset 40×) in infarction, inset shows macrophages

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