Notch pathway in ependymoma RELA-fused subgroup: upregulation and association with cancer stem cells markers expression

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Abstract

RELA-fused supratentorial (ST) ependymoma (EPN) is an aggressive subgroup with poor prognosis. Considering the putative role of Notch signaling in the maintenance of the cancer stem cells (CSC) phenotype in RELA-fused EPN, we investigated the expression of Notch pathway and its target genes in this subgroup. We also evaluated the effects of two Notch inhibitors (DAPT and RO4929097) on cell proliferation, apoptosis, colony formation, and CSCs markers gene expression on EPN cell line of the RELA-fused subgroup (BXD-1425). In addition, in silico signatures of the Notch genes and CSCs markers were analyzed on a large clinical dataset from GSE64415 study. We found that among the ST-EPN subgroups the Notch signaling (NOTCH1, JAG1, JAG2, and HES4) is specifically activated in the ST-EPN-RELA. Furthermore, treatment of the RELA-fused EPN cell line with the Notch inhibitors impaired the Notch signaling expression and revealed that Notch axis is not essential for cell proliferation and survival in this setting. NOTCH1 expression in ST-EPN was correlated with the CSCs markers VEGFA and L1CAM overexpression and JAG1 expression was correlated with the CCND1 and CDK6 overexpression. In addition, in vitro treatment with Notch inhibitors induced downregulation of CSCs markers. These findings indicate that Notch signaling can be involved in the ST-EPN-RELA CSCs maintenance by modulating the expression of genes responsible for cell phenotype and cell fate.

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Acknowledgements

We thank Xio Nan-Li that kindly provided BXD-1425 cell line for this study.

Funding

This collaborative study was supported by the Public Research Agencies: Fundação de Amparo a Pesquisa do Estado de São Paulo (FAPESP)—Grant nos. 2014/20341-0/Brazil and 2016/19820-6/ Brazil, Coordenação de Aperfeiçoamento de Pessoal de Nível Superior—Brasil (CAPES)—Finance Code 001, CNPq 457884/2014-2 and 151760/2018-7 and Fundação de Apoio ao Ensino, Pesquisa e Assistência do Hospital das Clínicas da Faculdade de Medicina de Ribeirão Preto da Universidade de São Paulo (FAEPA).

Author information

TAM planned and conducted all experiments, performed the in silico analysis, drafted and critically read the paper. GAVC helped to review the design of the study, performed the in silico analysis, and critically read the paper. GRS helped with RT-PCR and critically read the paper. KRS helped with in vitro assays and critically read the paper. RCPL helped to review the design of the study and critically read the paper. CAS critically read the paper. LGT critically read the paper. ETV helped with the design of the study and critically read the paper. KSB helped with the design of the study and critically read the paper.

Correspondence to Kleiton Silva Borges.

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