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The “go or grow” potential of gliomas is linked to the neuropeptide processing enzyme carboxypeptidase E and mediated by metabolic stress

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Abstract

Glioblastoma (GBM), the most common malignant brain tumor, is among the most lethal neoplasms, with a median survival of approximately 1 year. Prognosis is poor since GBMs possess a strong migratory and highly invasive potential, making complete surgical resection impossible. Reduced expression of carboxypeptidase E (CPE), a neuropeptide-processing enzyme, in a cell death-resistant glioma cell line and lower CPE expression levels in the cohort of GBM samples of The Cancer Genome Atlas compared to normal brain control specimens prompted us to analyze the function of CPE as a putative tumor suppressor gene. In our samples, CPE was also reduced in GBM compared to normal brain with the strongest loss in cells surrounding hypoxic tumor areas as well as in most glioma cell lines and primary glioma cells. In our cohort of glioma patients, loss of CPE predominantly occurred in glioblastomas and was associated with worse prognosis. In glioma cells, CPE overexpression was significantly reduced, whereas knockdown or inhibition enhanced glioma cell migration and invasion. The decreased migratory potential following CPE overexpression was paralleled by altered cellular morphology, promoting a transition to focal adhesions and associated stress fibers. In contrast to the decreased migration, high CPE levels were associated with higher proliferative rates. As microenvironmental regulation cues, we identified CPE as being downregulated upon hypoxia or glucose deprivation. Our findings indicate an oxygen- and nutrition-dependent anti-migratory, but pro-proliferative role of CPE in gliomas with prognostic impact for patient survival, thereby contributing to the understanding of the “go or grow” hypothesis in gliomas.

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Acknowledgments

The project was sponsored by the graduate program of the Interdisciplinary Centre for Clinical Research (IZKF) of the University of Tübingen.

Conflict of interest

All authors certify that there is no actual or potential conflict of interest in relation to this article.

Author information

Authors and Affiliations

  1. Laboratory of Molecular Neuro-Oncology, Department of General Neurology, Hertie Institute for Clinical Brain Research, University of Tübingen, Otfried-Müller-Str. 27, 72076, Tübingen, Germany

    Elisabeth Höring, Janina Seznec & Ulrike Naumann

  2. Institute of Neurology (Edinger-Institute), Goethe University of Frankfurt, Frankfurt/Main, Germany

    Patrick Nikolaus Harter, Cornelia Zachskorn & Michel Mittelbronn

  3. Department of Pathology and Neuropathology, University of Tübingen, Tübingen, Germany

    Jens Schittenhelm

  4. Laboratory for Stem Cell Research, Department of Internal Medicine II, University of Tübingen, Tübingen, Germany

    Hans-Jörg Bühring

  5. Graduate School for Cellular and Molecular Neuroscience, University of Tübingen, Tübingen, Germany

    Shohag Bhattacharyya

  6. Institute of Neuroradiology, Goethe University, Frankfurt/Main, Germany

    Elke von Hattingen

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  1. Elisabeth Höring
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  10. Ulrike Naumann
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Corresponding author

Correspondence to Ulrike Naumann.

Additional information

E. Höring and P. N. Harter contributed equally to this study.

Electronic supplementary material

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401_2011_940_MOESM1_ESM.tif

Supp. Fig. 1. CPE overexpressing (CPE no. 3) and control (neo no. 3) cells were lentivirally GFP transfected, cultured as hanging drop to create tumorspheres and placed on adult murine C57BL/6 brain slices. Tumor cell migration was tracked for 24 h by confocal live-cell imaging (using Nikon C1si confocal microscope, Nikon, Japan) (TIFF 15370 kb)

401_2011_940_MOESM2_ESM.tif

Supp. Fig. 2. CPE is downregulated in pseudopalisading glioma cells. Histology and immunohistochemistry of pseudopalisading cells. H&E staining showing central necrosis with pseudopalisading cells and vital rim (left). Immunohistochemistry for the proliferation marker Ki67 (middle) and CPE (right) in a higher magnification corresponding to the white frame of the H&E stain displays a marked reduction of CPE protein expression in areas with lower proliferating potential. (TIFF 12384 kb)

401_2011_940_MOESM3_ESM.tif

Supp. Fig. 3. CPE expression is not regulated on a transcriptional level. a CPE promoter analysis. NFκB Nuclear factor kappa B-binding element, CREB cyclic AMP response element (CRE)-binding protein consensus sequence, HRE HIF-1-binding, hypoxia-responsive element. b Regulation of CPE mRNA during hypoxia analyzed by real-time RT-PCR (n = 4, SEM). c Regulation of CPE mRNA during glucose deprivation analyzed as in b (n = 3, SEM). (TIFF 102026 kb)

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Höring, E., Harter, P.N., Seznec, J. et al. The “go or grow” potential of gliomas is linked to the neuropeptide processing enzyme carboxypeptidase E and mediated by metabolic stress. Acta Neuropathol 124, 83–97 (2012). https://doi.org/10.1007/s00401-011-0940-x

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  • DOI: https://doi.org/10.1007/s00401-011-0940-x

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