Glioblastoma (GBM) is associated with poor prognosis despite aggressive surgical resection, chemotherapy, and radiation therapy. Unfortunately, this standard therapy does not target glioma cancer stem cells (GCSCs), a subpopulation of GBM cells that can give rise to recurrent tumors. GBMs express human cytomegalovirus (HCMV) proteins, and previously we found that the level of expression of HCMV immediate-early (IE) protein in GBMs is a prognostic factor for poor patient survival. In this study, we investigated the relation between HCMV infection of GBM cells and the presence of GCSCs. Primary GBMs were characterized by their expression of HCMV-IE and GCSCs marker CD133 and by patient survival. The extent to which HCMV infection of primary GBM cells induced a GCSC phenotype was evaluated in vitro. In primary GBMs, a large fraction of CD133-positive cells expressed HCMV-IE, and higher co-expression of these two proteins predicted poor patient survival. Infection of GBM cells with HCMV led to upregulation of CD133 and other GSCS markers (Notch1, Sox2, Oct4, Nestin). HCMV infection also promoted the growth of GBM cells as neurospheres, a behavior typically displayed by GCSCs, and this phenotype was prevented by either chemical inhibition of the Notch1 pathway or by treatment with the anti-viral drug ganciclovir. GBM cells that maintained expression of HCMV-IE failed to differentiate into neuronal or astrocytic phenotypes. Our findings imply that HCMV infection induces phenotypic plasticity of GBM cells to promote GCSC features and may thereby increase the aggressiveness of this tumor.

Danish Cancer Society Research Center Copenhagen Denmark

Department of Medical Biochemistry and Biophysics Karolinska Institute Stockholm Sweden

Department of Neurology and Neurosurgery Karolinska University Hospital Stockholm Sweden

Department of Neurosurgery Copenhagen University Hospital Rigshospitalet Copenhagen Denmark

Institute of Molecular and Translational Medicine Faculty of Medicine and Dentistry Palacky University Olomouc Czech Republic

Karolinska Institute Department of Medicine Center for Molecular Medicine Cell and Molecular Immunology Microbial Pathogenesis Unit Stockholm Sweden

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1Wen PY, Kesari S. Malignant gliomas in adults. N Engl J Med 2008; 359: 492–507. PubMed

2Lima FR, Kahn SA, Soletti RC, Biasoli D, Alves T, da Fonseca AC et al. Glioblastoma: therapeutic challenges, what lies ahead. Biochim Biophys Acta 2012; 1826: 338–349. PubMed

3Singh SK, Hawkins C, Clarke ID, Squire JA, Bayani J, Hide T et al. Identification of human brain tumour initiating cells. Nature 2004; 432: 396–401. PubMed

4Cheng JX, Liu BL, Zhang X. How powerful is CD133 as a cancer stem cell marker in brain tumors? Cancer Treat Rev 2009; 35: 403–408. PubMed

5Ardebili SY, Zajc I, Gole B, Campos B, Herold-Mende C, Drmota S et al. CD133/prominin1 is prognostic for GBM patient's survival, but inversely correlated with cysteine cathepsins' expression in glioblastoma derived spheroids. Radiol Oncol 2011; 45: 102–115. PubMed PMC

6Zeppernick F, Ahmadi R, Campos B, Dictus C, Helmke BM, Becker N et al. Stem cell marker CD133 affects clinical outcome in glioma patients. Clin Cancer Res 2008; 14: 123–129. PubMed

7Beier D, Hau P, Proescholdt M, Lohmeier A, Wischhusen J, Oefner PJ et al. CD133(+) and CD133(-) glioblastoma-derived cancer stem cells show differential growth characteristics and molecular profiles. Cancer Res 2007; 67: 4010–4015. PubMed

8Ogden AT, Waziri AE, Lochhead RA, Fusco D, Lopez K, Ellis JA et al. Identification of A2B5+CD133- tumor-initiating cells in adult human gliomas. Neurosurgery 2008; 62: 505–514; discussion 514-505. PubMed

9Wang J, Sakariassen PO, Tsinkalovsky O, Immervoll H, Boe SO, Svendsen A et al. CD133 negative glioma cells form tumors in nude rats and give rise to CD133 positive cells. Int J Cancer 2008; 122: 761–768. PubMed

10Chen R, Nishimura MC, Bumbaca SM, Kharbanda S, Forrest WF, Kasman IM et al. A hierarchy of self-renewing tumor-initiating cell types in glioblastoma. Cancer Cell 2010; 17: 362–375. PubMed

11Ishiwata T, Teduka K, Yamamoto T, Kawahara K, Matsuda Y, Naito Z. Neuroepithelial stem cell marker nestin regulates the migration, invasion and growth of human gliomas. Oncol Rep 2011; 26: 91–99. PubMed

12Pannuti A, Foreman K, Rizzo P, Osipo C, Golde T, Osborne B et al. Targeting Notch to target cancer stem cells. Clin Cancer Res 2010; 16: 3141–3152. PubMed PMC

13Takahashi K, Yamanaka S. Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell 2006; 126: 663–676. PubMed

14Stopschinski BE, Beier CP, Beier D. Glioblastoma cancer stem cells—from concept to clinical application. Cancer Lett 2013; 338: 32–40. PubMed

15Florio T, Barbieri F. The status of the art of human malignant glioma management: the promising role of targeting tumor-initiating cells. Drug Disc Today 2012; 17: 1103–1110. PubMed

16Cobbs CS, Harkins L, Samanta M, Gillespie GY, Bharara S, King PH et al. Human cytomegalovirus infection and expression in human malignant glioma. Cancer Res 2002; 62: 3347–3350. PubMed

17Soderberg-Naucler C, Rahbar A, Stragliotto G. Survival in patients with glioblastoma receiving valganciclovir. N Engl J Med 2013; 369: 985–986. PubMed

18Rahbar A, Orrego A, Peredo I, Dzabic M, Wolmer-Solberg N, Straat K et al. Human cytomegalovirus infection levels in glioblastoma multiforme are of prognostic value for survival. J Clin Virol 2013; 57: 36–42. PubMed

19Sinclair J. Human cytomegalovirus: Latency and reactivation in the myeloid lineage. J Clin Virol 2008; 41: 180–185. PubMed

20Soderberg-Naucler C, Fish KN, Nelson JA. Reactivation of latent human cytomegalovirus by allogeneic stimulation of blood cells from healthy donors. Cell 1997; 91: 119–126. PubMed

21Perez-Garcia A, Carrion-Navarro J, Bosch-Fortea M, Lazaro-Ibanez E, Prat-Acin R, Ayuso-Sacido A. Genomic instability of surgical sample and cancer-initiating cell lines from human glioblastoma. Front Biosci 2012; 17: 1469–1479. PubMed

22Bartkova J, Hamerlik P, Stockhausen MT, Ehrmann J, Hlobilkova A, Laursen H et al. Replication stress and oxidative damage contribute to aberrant constitutive activation of DNA damage signalling in human gliomas. Oncogene 2010; 29: 5095–5102. PubMed

23Verhaak RG, Hoadley KA, Purdom E, Wang V, Qi Y, Wilkerson MD et al. Integrated genomic analysis identifies clinically relevant subtypes of glioblastoma characterized by abnormalities in PDGFRA, IDH1, EGFR, and NF1. Cancer Cell 2010; 17: 98–110. PubMed PMC

24Patel AP, Tirosh I, Trombetta JJ, Shalek AK, Gillespie SM, Wakimoto H et al. Single-cell RNA-seq highlights intratumoral heterogeneity in primary glioblastoma. Science 2014; 344: 1396–1401. PubMed PMC

25Brescia P, Ortensi B, Fornasari L, Levi D, Broggi G, Pelicci G. CD133 is essential for glioblastoma stem cell maintenance. Stem Cell 2013; 31: 857–869. PubMed

26Odeberg J, Wolmer N, Falci S, Westgren M, Seiger A, Soderberg-Naucler C. Human cytomegalovirus inhibits neuronal differentiation and induces apoptosis in human neural precursor cells. J Virol 2006; 80: 8929–8939. PubMed PMC

27Hu J, Ho AL, Yuan L, Hu B, Hua S, Hwang SS et al. From the Cover: Neutralization of terminal differentiation in gliomagenesis. Proc Natl Acad Sci USA 2013; 110: 14520–14527. PubMed PMC

28Ranganathan P, Clark PA, Kuo JS, Salamat MS, Kalejta RF. Significant association of multiple human cytomegalovirus genomic Loci with glioblastoma multiforme samples. J Virol 2012; 86: 854–864. PubMed PMC

29Bhattacharjee B, Renzette N, Kowalik TF. Genetic analysis of cytomegalovirus in malignant gliomas. J Virol 2012; 86: 6815–6824. PubMed PMC

30Mitchell DA, Xie W, Schmittling R, Learn C, Friedman A, McLendon RE et al. Sensitive detection of human cytomegalovirus in tumors and peripheral blood of patients diagnosed with glioblastoma. Neuro-oncol 2008; 10: 10–18. PubMed PMC

31Hadaczek P, Ozawa T, Soroceanu L, Yoshida Y, Matlaf L, Singer E et al. Cidofovir: a novel antitumor agent for glioblastoma. Clin Cancer Res 2013; 19: 6473–6483. PubMed PMC

32Stern-Ginossar N, Weisburd B, Michalski A, Le VT, Hein MY, Huang SX et al. Decoding human cytomegalovirus. Science 2012; 338: 1088–1093. PubMed PMC

33Fan X, Khaki L, Zhu TS, Soules ME, Talsma CE, Gul N et al. NOTCH pathway blockade depletes CD133-positive glioblastoma cells and inhibits growth of tumor neurospheres and xenografts. Stem Cell 2010; 28: 5–16. PubMed PMC

34Venere M, Hamerlik P, Wu Q, Rasmussen RD, Song LA, Vasanji A et al. Therapeutic targeting of constitutive PARP activation compromises stem cell phenotype and survival of glioblastoma-initiating cells. Cell Death Differ 2014; 21: 258–269. PubMed PMC