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EZH2 inhibitory protein (EZHIP/Cxorf67) expression correlates strongly with H3K27me3 loss in posterior fossa ependymomas and is mutually exclusive with H3K27M mutations

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A Correction to this article was published on 09 January 2021

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Abstract

The PFA molecular subgroup of posterior fossa ependymomas (PF-EPNs) shows poor outcome. H3K27me3 (me3) loss by immunohistochemistry (IHC) is a surrogate marker for PFA wherein its loss is attributed to overexpression of Cxorf67/EZH2 inhibitory protein (EZHIP), C17orf96, and ATRX loss. We aimed to subgroup PF-EPNs using me3 IHC and study correlations of the molecular subgroups with other histone related proteins, 1q gain, Tenascin C and outcome. IHC for me3, acetyl-H3K27, H3K27M, ATRX, EZH2, EZHIP, C17orf96, Tenascin-C, and fluorescence in-situ hybridisation for chromosome 1q25 locus were performed on an ambispective PF-EPN cohort (2003–2019). H3K27M-mutant gliomas were included for comparison. Among 69 patients, PFA (me3 loss) constituted 64%. EZHIP overexpression and 1q gain were exclusive to PFA seen in 72% and 19%, respectively. Tenascin C was more frequently positive in PFA (p = 0.02). H3K27M expression and ATRX loss were noted in one case of PFA–EPN each. All H3K27M-mutant gliomas (n = 8) and PFA-EPN (n = 1) were EZHIP negative. C17orf96 and acetyl-H3K27 expression did not correlate with me3 loss. H3K27me3 is a robust surrogate for PF-EPN molecular subgrouping. EZHIP overexpression was exclusive to PFA EPNs and was characteristically absent in midline gliomas and the rare PFA harbouring H3K27M mutations representing mutually exclusive pathways leading to me3 loss.

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  • 09 January 2021

    In the original publication of the article, the middle name was missing in corresponding author’s name.

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Acknowledgements

This work is supported by Central Institute of Industrial research (CSIR), New Delhi, India (Pool No 8948A/17), and Neurosciences centre, AIIMS, New Delhi.

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Authors and Affiliations

  1. Department of Pathology, All India Institute of Medical Sciences, Ansari Nagar, New Delhi, 110029, India

    Aruna Nambirajan, Agrima Sharma, Madhu Rajeshwari, Vaishali Suri, Chitra Sarkar & Mehar Chand Sharma

  2. Department of Neurosurgery, All India Institute of Medical Sciences, New Delhi, India

    Meher Tej Boorgula & Ramesh Doddamani

  3. Department of Neuroradiology, All India Institute of Medical Sciences, New Delhi, India

    Ajay Garg

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The original online version of this article was revised due to the name of corresponding author was published incorrectly and corrected in this version.

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10014_2020_385_MOESM1_ESM.tif

Supplementary Figure 1: Tukey plot showing EZH2 scores among different subgroups of PF-EPNs and H3K27M-mutant brain stem DMG in the study. *Significant difference in score was seen only between EZHIP positive and negative PF-EPNs and none of the other sub groups (TIF 17 kb)

10014_2020_385_MOESM2_ESM.tif

Supplementary Figure 2: Histomorphology and immunohistochemical features of the H3K27M-mutant PFA ependymoma. The histomorphology was that of a classical Grade 2 ependymoma with perivascular pseudo-rosettes (A) and showed characteristic dot-like staining for epithelial membrane antigen (B). There was loss of staining for H3K27me3 in tumour cells (C) with strong diffuse staining for H3K27M protein (D). Sanger sequencing chromatogram shows presence of H3F3A p.K27M (orange arrows) in tumour cells using reverse (E) and forward (F) primers (TIF 4016 kb)

10014_2020_385_MOESM3_ESM.tif

Supplementary Figure 3: Histomorphology and immunohistochemical features of H3K27M-mutant diffuse midline gliomas. A representative case showing histomorphology of an infiltrating glioma (A) showing loss of H3K27me3 staining in tumour cells (B) with strong diffuse staining for H3K27M protein (D). EZH2 expression was variable (E) while there was diffuse staining for acetyl-H3K27 (F) (TIF 6007 kb)

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Nambirajan, A., Sharma, A., Rajeshwari, M. et al. EZH2 inhibitory protein (EZHIP/Cxorf67) expression correlates strongly with H3K27me3 loss in posterior fossa ependymomas and is mutually exclusive with H3K27M mutations. Brain Tumor Pathol 38, 30–40 (2021). https://doi.org/10.1007/s10014-020-00385-9

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