P20.10.B CHARACTERIZATION OF NEW PROGNOSTIC MARKERS IN A COHORT OF ADULT SPINAL EPENDYMOMAS
Authors: Patanè M , Calatozzolo C , Maddaloni L , Paterra R , Marucci G , Eoli M , Farinotti M , Ferroli P , Silvani A , Pollo B 
Published: September 2023

Abstract

BACKGROUND
Spinal ependymomas (SP-EPN) are group of primary central nervous system tumors recently re-classified by the WHO (CNS5 2021), located in spinal cord. SP-EPN are grade 2-3 tumors, found mainly in cervical and thoracic region, characterized mostly by NF2 mutations and classical histology: grade 3 tumors present also microvascular proliferation, necrosis and high mitotic rate. Recently, a novel subtype was discovered with a unique methylation profile (SP-EPN-MYCN) and characterized by the amplification of MYCN gene, grade 3 histology and a very aggressive behavior. Treatment of these tumors remains challenging: gross-total resection is not often achieved and radiation failed to give sacceptable results. Besides, no prognostic factors except the mitotic index and tumor location, are available, thus, new diagnostic and prognostic biomarkers would be very useful for their accurate classification and management.

MATERIAL AND METHODS
We have collected 170 patients operated at our Institute from 1997 to 2019 of spinal ependymomas located in the cervical and thoracic region, excluded conus/filum mixopapillary ones. All tumors were revised according to the new WHO CNS5 2021 classification. In a cohort of 50 grade 2/3 SP-EPN we have evaluated by immunohistochemistry the expression of MYCN, H3K27me3, CDK4 and EZH2: the latter is a component of the PRC2 complex, which regulates the histone H3 methylation at Lys27 (K27) and recently described as a transcription factor which controls the expression of MYCN and CDK4 genes. We have also evaluated the amplification of MYCN and CDK4 genes by Copy Number Variation Assay and MLPA techniques. Clinical data were collected and related to the results obtained.

RESULTS
We have found strong overexpression of MYCN only in grade 3 SP-EPN (3/50), while grade 2 tumors showed low or no expression; amplification of MYCN gene was similarly confirmed by CNV assay and MLPA only in grade 3. Interestingly, also CDK4 overexpression was found only in grade 3 ependymomas (1/50), but it is expressed oppositely to MYCN: in tumors were MYCN is overexpressed, CDK4 was completely absent. Amplification of CDK4 gene was confirmed only in those with CDK4 overexpression. These results were both related to an interesting overexpression of EZH2 only in grade 3 SP-EPN (4/50), while H3K27me3 is expressed in all grades without differences. Finally, in our study both MYCN and CDK4-amplified ependymomas were related to a very aggressive behaviour and poor outcome with high recurrence rate, dissemination and metastases.

CONCLUSION
In this study we confirmed the prognostic role of MYCN as indicated by WHO classification and we have showed that CDK4 and EZH2 overexpression could be considered as new promising prognostic and predictive factors in SP-EPN. Since many trials with EZH2 and CDK4 inhibitors have been set up in the last years, these analyses could suggest a new therapeutic approach.