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1. Cellular heterogeneity contributes to subtype-specific expression of ZEB1 in human glioblastoma.

Author

Philipp Euskirchen, Josefine Radke, Marc Sören Schmidt, Eva Schulze Heuling, Eric Kadikowski, Meron Maricos, Felix Knab, Ulrike Grittner, Norman Zerbe, Marcus Czabanka, Christoph Dieterich, Hrvoje Miletic, Sverre Mørk, Arend Koch, Matthias Endres, and Christoph Harms

Subjects
Medicine, Science
Abstract

The transcription factor ZEB1 has gained attention in tumor biology of epithelial cancers because of its function in epithelial-mesenchymal transition, DNA repair, stem cell biology and tumor-induced immunosuppression, but its role in gliomas with respect to invasion and prognostic value is controversial. We characterized ZEB1 expression at single cell level in 266 primary brain tumors and present a comprehensive dataset of high grade gliomas with Ki67, p53, IDH1, and EGFR immunohistochemistry, as well as EGFR FISH. ZEB1 protein expression in glioma stem cell lines was compared to their parental tumors with respect to gene expression subtypes based on RNA-seq transcriptomic profiles. ZEB1 is widely expressed in glial tumors, but in a highly variable fraction of cells. In glioblastoma, ZEB1 labeling index is higher in tumors with EGFR amplification or IDH1 mutation. Co-labeling studies showed that tumor cells and reactive astroglia, but not immune cells contribute to the ZEB1 positive population. In contrast, glioma cell lines constitutively express ZEB1 irrespective of gene expression subtype. In conclusion, our data indicate that immune infiltration likely contributes to differential labelling of ZEB1 and confounds interpretation of bulk ZEB1 expression data.

Published
2017
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2. Cellular heterogeneity contributes to subtype-specific expression of ZEB1 in human glioblastoma

Author

Eva Schulze Heuling, Marc Soeren Schmidt, Matthias Endres, Arend Koch, Christoph Harms, Philipp Euskirchen, Christoph Dieterich, Marcus Czabanka, Sverre Mørk, Norman Zerbe, Meron Maricos, Ulrike Grittner, Eric Kadikowski, Felix Knab, Josefine Radke, and Hrvoje Miletic

Subjects
Microarrays, 600 Technik, Medizin, angewandte Wissenschaften::610 Medizin und Gesundheit, Transcriptome, Animal Cells, Gene expression, Blastomas, RNA, Neoplasm, lcsh:Science, Neurological Tumors, Cultured Tumor Cells, Mutation, education.field_of_study, Brain Neoplasms, Immunohistochemistry, ErbB Receptors, Bioassays and Physiological Analysis, Oncology, Neurology, metabolism [Zinc Finger E-box-Binding Homeobox 1], Biological Cultures, Cellular Types, metabolism [ErbB Receptors], Immune Cells, Immunology, IDH1 protein, human, genetics [Isocitrate Dehydrogenase], 03 medical and health sciences, Cancer stem cell, Glioma, Genetics, Humans, RNA, Messenger, ZEB1 protein, human, education, Blood Cells, Macrophages, lcsh:R, metabolism [RNA, Neoplasm], Zinc Finger E-box-Binding Homeobox 1, Biology and Life Sciences, medicine.disease, genetics [Brain Neoplasms], 030104 developmental biology, metabolism [Isocitrate Dehydrogenase], lcsh:Q, Glioblastoma, pathology [Glioma], 0301 basic medicine, genetics [Glioma], lcsh:Medicine, Gene Expression, genetics [Glioblastoma], medicine.disease_cause, metabolism [Glioma], pathology [Glioblastoma], White Blood Cells, Tumor Microenvironment, Medicine and Health Sciences, genetics [RNA, Neoplasm], In Situ Hybridization, Fluorescence, Staining, genetics [Zinc Finger E-box-Binding Homeobox 1], Multidisciplinary, Cell Staining, Isocitrate Dehydrogenase, metabolism [Brain Neoplasms], Research Article, pathology [Brain Neoplasms], DNA repair, Population, Biology, Research and Analysis Methods, metabolism [RNA, Messenger], genetics [ErbB Receptors], genetics [RNA, Messenger], EGFR protein, human, Cell Line, Tumor, immunology [Tumor Microenvironment], medicine, ddc:610, Immunohistochemistry Techniques, Cancers and Neoplasms, Cell Biology, Cell Cultures, genetics [Tumor Microenvironment], Glioma Cells, Molecular biology, Histochemistry and Cytochemistry Techniques, Specimen Preparation and Treatment, Cell culture, Immunologic Techniques, Cancer research, metabolism [Glioblastoma], Glioblastoma Multiforme
Abstract

The transcription factor ZEB1 has gained attention in tumor biology of epithelial cancers because of its function in epithelial-mesenchymal transition, DNA repair, stem cell biology and tumor-induced immunosuppression, but its role in gliomas with respect to invasion and prognostic value is controversial. We characterized ZEB1 expression at single cell level in 266 primary brain tumors and present a comprehensive dataset of high grade gliomas with Ki67, p53, IDH1, and EGFR immunohistochemistry, as well as EGFR FISH. ZEB1 protein expression in glioma stem cell lines was compared to their parental tumors with respect to gene expression subtypes based on RNA-seq transcriptomic profiles. ZEB1 is widely expressed in glial tumors, but in a highly variable fraction of cells. In glioblastoma, ZEB1 labeling index is higher in tumors with EGFR amplification or IDH1 mutation. Co-labeling studies showed that tumor cells and reactive astroglia, but not immune cells contribute to the ZEB1 positive population. In contrast, glioma cell lines constitutively express ZEB1 irrespective of gene expression subtype. In conclusion, our data indicate that immune infiltration likely contributes to differential labelling of ZEB1 and confounds interpretation of bulk ZEB1 expression data. publishedVersion

Published
2017

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