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Fibroblast activation protein alpha is expressed by transformed and stromal cells and is associated with mesenchymal features in glioblastoma

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Tumor Biology

Abstract

Glioblastomas are deadly neoplasms resistant to current treatment modalities. Fibroblast activation protein (FAP) is a protease which is not expressed in most of the normal adult tissues but is characteristically present in the stroma of extracranial malignancies. FAP is considered a potential therapeutic target and is associated with a worse patient outcome in some cancers. The FAP localization in the glioma microenvironment and its relation to patient survival are unknown. By analyzing 56 gliomas and 15 non-tumorous brain samples, we demonstrate increased FAP expression in a subgroup of high-grade gliomas, in particular on the protein level. FAP expression was most elevated in the mesenchymal subtype of glioblastoma. It was neither associated with glioblastoma patient survival in our patient cohort nor in publicly available datasets. FAP was expressed in both transformed and stromal cells; the latter were frequently localized around dysplastic blood vessels and commonly expressed mesenchymal markers. In a mouse xenotransplantation model, FAP was expressed in glioma cells in a subgroup of tumors that typically did not express the astrocytic marker GFAP. Endogenous FAP was frequently upregulated and part of the FAP+ host cells coexpressed the CXCR4 chemokine receptor. In summary, FAP is expressed by several constituents of the glioblastoma microenvironment, including stromal non-malignant mesenchymal cells recruited to and/or activated in response to glioma growth. The limited expression of FAP in healthy tissues together with its presence in both transformed and stromal cells suggests that FAP may be a candidate target for specific delivery of therapeutic agents in glioblastoma.

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Acknowledgments

The work was supported by the grant from the Ministry of Health of the Czech Republic, Internal grant agency IGA 12237-5/2011. The authors thank Igor Romanko for assistance with CXCR4 immunohistochemistry and Zdislava Vanickova for the help with RNA isolation. The excellent technical assistance of Kvetoslava Vlasicova and Karin Roubickova is acknowledged. Special thanks go to Martin Prevorovsky for the help with the TCGA expression data.

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Authors and Affiliations

  1. Institute of Biochemistry and Experimental Oncology, First Faculty of Medicine, Charles University in Prague, U Nemocnice 5, 128 53, Prague 2, Czech Republic

    Petr Busek, Eva Balaziova, Ivana Matrasova, Marek Hilser, Evzen Krepela & Aleksi Sedo

  2. Department of Neurosurgery, Na Homolce Hospital, Roentgenova 2, 150 30, Prague 5, Czech Republic

    Robert Tomas

  3. Department of Pathology, Na Homolce Hospital, Roentgenova 2, 150 30, Prague 5, Czech Republic

    Martin Syrucek

  4. Institute of Clinical Biochemistry and Laboratory Diagnostics of the First Faculty of Medicine, Charles University in Prague and General University Hospital in Prague, U Nemocnice 2, 128 01, Prague 2, Czech Republic

    Zuzana Zemanova

  5. Institute of Biophysics and Bioinformatics, First Faculty of Medicine, Charles University in Prague, Salmovská 1, 120 00, Prague 2, Czech Republic

    Jaromir Belacek

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  1. Petr Busek
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  2. Eva Balaziova
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  3. Ivana Matrasova
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  10. Aleksi Sedo
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Correspondence to Petr Busek or Aleksi Sedo.

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Funding

This study was funded by the Ministry of Health of the Czech Republic, Internal grant agency IGA 12237–5/2011.

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Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards. All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.

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Supplementary Figure S1

Characteristics of the glioma stem-like cell cultures a) GSC57 and b) GSC48. Expression of Sox2 (Sox2-PerCP, RD Systems) and CD133 (CD133-APC, Miltenyi Biotech) was determined by flow cytometry, detection of the differentiation markers glial fibrillary acidic protein (GFAP) and beta III tubulin was performed in cells grown on laminin in defined stem-cell serum free media (undifferentiated) or in the presence of 10 % fetal calf serum (differentiated). (PPTX 785 kb)

Supplementary Figure S2

Fibroblast activation protein (FAP) and survival in gliomas. The Kaplan-Meier survival plots for glioma patients and the proportion of individual tumor grades in the subgroups based on FAP expression in studies comprising grade I-IV tumors. A) Greavendeel [9] dataset, b) Data from REMBRANDT. Patients were divided into groups based on FAP mRNA expression: ∆ low expression = 1st quartile, □ medium expression = 2nd + 3rd quartile, ○ high expression = 4th quartile. + = censored data. Log rank test p < 0.05 for all intergroup comparisons in A), and p < 0.05 for downregulated vs. intermediate and downregulated vs. upregulated in B). (PPTX 146 kb)

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Busek, P., Balaziova, E., Matrasova, I. et al. Fibroblast activation protein alpha is expressed by transformed and stromal cells and is associated with mesenchymal features in glioblastoma. Tumor Biol. 37, 13961–13971 (2016). https://doi.org/10.1007/s13277-016-5274-9

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