miRNA-26a expression influences the therapy response to carmustine wafer implantation in patients with glioblastoma multiforme
Glioblastoma multiforme is the most frequent malignant brain tumor in adults being marked with a very poor prognosis. Therapy concept implies concomitant radio-chemotherapy and facultative implantation of carmustine-eluted wafer. Current literature suggests microRNA 26a expression in glioblastoma to interact with alkylating chemotherapy. Subsequently, the aim of this study was to investigate the correlation of miRNA-26a expression and carmustine wafer implantation and its potential usefulness as a predictive marker for therapy response.
In total, 229 patients with glioblastoma multiforme were included into the final analysis. Of them, 80 cases were recruited from the Saarland University Medical Center for a retrospective matched-pair analysis stratified after therapy regime: One group (carmustine wafer group; n=40) received concomitant radio-chemotherapy with carmustine wafer implantation. The other group (control group; n=40) only received concomitant radio-chemotherapy. The results were confirmed by comparing them with an independent dataset of 149 patients from the TCGA database. All tumor specimens were evaluated for miRNA-26a expression, MGMT promoter methylation, and IDH1 R132H mutation status, and the results were correlated with the clinical data.
Twenty-three patients in the carmustine wafer group showed low expression of miRNA-26a, while 17 patients showed a high expression. In the control group, 28 patients showed low expression, while 12 patients showed a high expression. The patients with high miRNA-26a expression in the carmustine wafer group were characterized by a significantly longer overall (hazard ratio [HR] 2.750 [95% CI 1.352–5.593]; p=0.004) and progression-free survival (HR 3.091 [95% CI 1.436–6.657]; p=0.003) than patients with low miRNA-26a expression. The 17 patients in the carmustine wafer group with high miRNA-26a expression showed a significantly longer progression-free survival (p=0.013) and overall survival (p=0.007) compared with the control group. There were no such correlations identified within the control group. TCGA datasets supported these findings.
MiRNA-26a expression turned out to be a promising predictor of therapy response and clinical outcome in glioblastoma patients treated with carmustine wafer implantation. For evaluation of the role of miRNA-26a in a combined therapy setting, further studies are needed in order to translate general findings to the patient’s individual situation.
KeywordsmiRNA-26a Carmustine wafer Epigenetic Glioblastoma
Gross total resection
Karnofsky Performance Score
Magnetic resonance imaging
Methylation-specific polymerase chain reaction
Quantitative reverse-transcription polymerase chain reaction
The Cancer Genome Atlas
The authors wish to thank Lisa Senger for her editorial support and Sigrid Welsch for their technical assistance in miRNA and methylation analysis.
This study was financed by Archimedes Pharma (L204150209).
Compliance with ethical standards
All procedures performed in this study were in accordance with the ethical standards of the 1964 Helsinki declaration. This article does not contain any studies with animals performed by any of the authors.
Ethics approval and consent to participate
This study was approved by the local German ethical board (Ethikkommission der Ärztekammer des Saarlandes, Saarbrücken, Germany).
Consent for publication
Written informed consent was obtained from all patients (General Medical Council of the State of Saarland, NO 93/16).
The authors declare that they have no competing interests.
- 1.Abd-El-Barr MM, Chiocca EA (2014) How much is enough? The question of extent of resection in glioblastoma multiforme. World Neurosurg 82(1–2):e109–e110Google Scholar
- 2.Ahir BK, Ozer H, Engelhard HH, Lakka SS (2017) MicroRNAs in glioblastoma pathogenesis and therapy: a comprehensive review. Crit Rev Oncol Hematol 120:22–33Google Scholar
- 3.Bäcklund LM, Nilsson BR, Goike HM, Schmidt EE, Liu L, Ichimura K, Collins VP (2003) Short postoperative survival for glioblastoma patients with a dysfunctional Rb1 pathway in combination with no wild-type PTEN. Clin Cancer Res Off J Am Assoc Cancer Res 9(11):4151–4158Google Scholar
- 4.Bäcklund LM, Nilsson BR, Liu L, Ichimura K, Collins VP (2005) Mutations in Rb1 pathway-related genes are associated with poor prognosis in anaplastic astrocytomas. Br J Cancer 93(1):124–130Google Scholar
- 5.Bartel DP (2004) MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 116(2):281–297Google Scholar
- 6.Busato F, Dejeux E, El Abdalaoui H, Gut IG, Tost J (2018) Quantitative DNA methylation analysis at single-nucleotide resolution by pyrosequencing®. Methods Mol Biol Clifton NJ 1708:427–445Google Scholar
- 7.Cancer Genome Atlas Research Network (2008) Comprehensive genomic characterization defines human glioblastoma genes and core pathways. Nature 455(7216):1061–1068Google Scholar
- 8.Capper D, Weissert S, Balss J et al (2010) Characterization of R132H mutation-specific IDH1 antibody binding in brain tumors. Brain Pathol Zurich Switz 20(1):245–254Google Scholar
- 9.Chaichana KL, Cabrera-Aldana EE, Jusue-Torres I, Wijesekera O, Olivi A, Rahman M, Quinones-Hinojosa A (2014) When gross total resection of a glioblastoma is possible, how much resection should be achieved? World Neurosurg 82(1–2):e257–e265Google Scholar
- 10.Chen C-Z (2005) MicroRNAs as oncogenes and tumor suppressors. N Engl J Med 353(17):1768–1771Google Scholar
- 11.Colaprico A, Silva TC, Olsen C et al (2016) TCGAbiolinks: an R/Bioconductor package for integrative analysis of TCGA data. Nucleic Acids Res 44(8):e71Google Scholar
- 12.Curran WJ, Scott CB, Horton J, Nelson JS, Weinstein AS, Fischbach AJ, Chang CH, Rotman M, Asbell SO, Krisch RE (1993) Recursive partitioning analysis of prognostic factors in three Radiation Therapy Oncology Group malignant glioma trials. J Natl Cancer Inst 85(9):704–710Google Scholar
- 13.De Bonis P, Anile C, Pompucci A, Fiorentino A, Balducci M, Chiesa S, Maira G, Mangiola A (2012) Safety and efficacy of Gliadel wafers for newly diagnosed and recurrent glioblastoma. Acta Neurochir 154(8):1371–1378Google Scholar
- 14.Felsberg J, Rapp M, Loeser S, Fimmers R, Stummer W, Goeppert M, Steiger H-J, Friedensdorf B, Reifenberger G, Sabel MC (2009) Prognostic significance of molecular markers and extent of resection in primary glioblastoma patients. Clin Cancer Res Off J Am Assoc Cancer Res 15(21):6683–6693Google Scholar
- 15.Ge X, Pan M-H, Wang L, Li W, Jiang C, He J, Abouzid K, Liu L-Z, Shi Z, Jiang B-H (2018) Hypoxia-mediated mitochondria apoptosis inhibition induces temozolomide treatment resistance through miR-26a/Bad/Bax axis. Cell Death Dis 9(11):1128Google Scholar
- 16.Gil J, Peters G (2006) Regulation of the INK4b-ARF-INK4a tumour suppressor locus: all for one or one for all. Nat Rev Mol Cell Biol 7(9):667–677Google Scholar
- 17.Goldhoff P, Clarke J, Smirnov I, Berger MS, Prados MD, James CD, Perry A, Phillips JJ (2012) Clinical stratification of glioblastoma based on alterations in retinoblastoma tumor suppressor protein (RB1) and association with the proneural subtype. J Neuropathol Exp Neurol 71(1):83–89Google Scholar
- 18.Grabowski MM, Recinos PF, Nowacki AS, Schroeder JL, Angelov L, Barnett GH, Vogelbaum MA (2014) Residual tumor volume versus extent of resection: predictors of survival after surgery for glioblastoma. J Neurosurg 121(5):1115–1123Google Scholar
- 19.Grossman SA, Ye X, Piantadosi S, Desideri S, Nabors LB, Rosenfeld M, Fisher J, NABTT CNS Consortium (2010) Survival of patients with newly diagnosed glioblastoma treated with radiation and temozolomide in research studies in the United States. Clin Cancer Res Off J Am Assoc Cancer Res 16(8):2443–2449Google Scholar
- 20.Halani SH, Babu R, Adamson DC (2017) Management of glioblastoma multiforme in the elderly: a review of the literature. World Neurosurg. https://doi.org/10.1016/j.wneu.2017.04.153
- 21.Hartmann C, Meyer J, Balss J et al (2009) Type and frequency of IDH1 and IDH2 mutations are related to astrocytic and oligodendroglial differentiation and age: a study of 1,010 diffuse gliomas. Acta Neuropathol (Berl) 118(4):469–474Google Scholar
- 22.Hegi ME, Diserens A-C, Gorlia T et al (2005) MGMT gene silencing and benefit from temozolomide in glioblastoma. N Engl J Med 352(10):997–1003Google Scholar
- 23.Hegi ME, Liu L, Herman JG, Stupp R, Wick W, Weller M, Mehta MP, Gilbert MR (2008) Correlation of O6-methylguanine methyltransferase (MGMT) promoter methylation with clinical outcomes in glioblastoma and clinical strategies to modulate MGMT activity. J Clin Oncol Off J Am Soc Clin Oncol 26(25):4189–4199Google Scholar
- 24.Henriksen M, Johnsen KB, Olesen P, Pilgaard L, Duroux M (2014) MicroRNA expression signatures and their correlation with clinicopathological features in glioblastoma multiforme. NeuroMolecular Med 16(3):565–577Google Scholar
- 25.Herman JG, Graff JR, Myöhänen S, Nelkin BD, Baylin SB (1996) Methylation-specific PCR: a novel PCR assay for methylation status of CpG islands. Proc Natl Acad Sci U S A 93(18):9821–9826Google Scholar
- 26.Holt DE, Bernard ME, Quan K, Clump DA, Engh JA, Burton SA, Heron DE (2016) Salvage stereotactic radiosurgery for recurrent glioblastoma multiforme with prior radiation therapy. J Cancer Res Ther 12(4):1243–1248Google Scholar
- 27.Huse JT, Brennan C, Hambardzumyan D et al (2009) The PTEN-regulating microRNA miR-26a is amplified in high-grade glioma and facilitates gliomagenesis in vivo. Genes Dev 23(11):1327–1337Google Scholar
- 28.Limentani SA, Asher A, Heafner M, Kim JW, Fraser R (2005) A phase I trial of surgery, Gliadel wafer implantation, and immediate postoperative carboplatin in combination with radiation therapy for primary anaplastic astrocytoma or glioblastoma multiforme. J Neuro-Oncol 72(3):241–244Google Scholar
- 29.Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods San Diego Calif 25(4):402–408Google Scholar
- 30.López-Urrutia E, Coronel-Hernández J, García-Castillo V et al (2017) MiR-26a downregulates retinoblastoma in colorectal cancer. Tumour Biol J Int Soc Oncodevelopmental Biol Med 39(4):1010428317695945Google Scholar
- 31.Louis DN, Perry A, Reifenberger G, von Deimling A, Figarella-Branger D, Cavenee WK, Ohgaki H, Wiestler OD, Kleihues P, Ellison DW (2016) The 2016 World Health Organization classification of tumors of the central nervous system: a summary. Acta Neuropathol (Berl) 131(6):803–820Google Scholar
- 32.Malzkorn B, Wolter M, Liesenberg F, Grzendowski M, Stühler K, Meyer HE, Reifenberger G (2010) Identification and functional characterization of microRNAs involved in the malignant progression of gliomas. Brain Pathol Zurich Switz 20(3):539–550Google Scholar
- 33.Nakamura M, Yonekawa Y, Kleihues P, Ohgaki H (2001) Promoter hypermethylation of the RB1 gene in glioblastomas. Lab Investig J Tech Methods Pathol 81(1):77–82Google Scholar
- 34.Nikolova T, Roos WP, Krämer OH, Strik HM, Kaina B (2017) Chloroethylating nitrosoureas in cancer therapy: DNA damage, repair and cell death signaling. Biochim Biophys Acta Rev Cancer 1868(1):29–39Google Scholar
- 35.Ostrom QT, Gittleman H, Farah P, Ondracek A, Chen Y, Wolinsky Y, Stroup NE, Kruchko C, Barnholtz-Sloan JS (2013) CBTRUS statistical report: primary brain and central nervous system tumors diagnosed in the United States in 2006-2010. Neuro-Oncol 15(Suppl 2):ii1–i56Google Scholar
- 36.Pallud J, Audureau E, Noel G et al (2015) Long-term results of carmustine wafer implantation for newly diagnosed glioblastomas: a controlled propensity-matched analysis of a French multicenter cohort. Neuro-Oncol 17(12):1609–1619Google Scholar
- 37.Parker NR, Correia N, Crossley B, Buckland ME, Howell VM, Wheeler HR (2013) Correlation of microRNA 132 up-regulation with an unfavorable clinical outcome in patients with primary glioblastoma multiforme treated with radiotherapy plus concomitant and adjuvant temozolomide chemotherapy. Transl Oncol 6(6):742–748Google Scholar
- 38.Sabel M, Giese A (2008) Safety profile of carmustine wafers in malignant glioma: a review of controlled trials and a decade of clinical experience. Curr Med Res Opin 24(11):3239–3257Google Scholar
- 39.Sawaya R (1999) Extent of resection in malignant gliomas: a critical summary. J Neuro-Oncol 42(3):303–305Google Scholar
- 40.Simpson DJ, Hibberts NA, McNicol AM, Clayton RN, Farrell WE (2000) Loss of pRb expression in pituitary adenomas is associated with methylation of the RB1 CpG island. Cancer Res 60(5):1211–1216Google Scholar
- 41.Sippl C, Ketter R, Bohr L, Kim YJ, List M, Oertel J, Urbschat S (2018) MiRNA-181d expression significantly affects treatment responses to carmustine wafer implantation. Neurosurgery. https://doi.org/10.1093/neuros/nyy214
- 42.Sippl C, Urbschat S, Kim YJ, Senger S, Oertel J, Ketter R (2018) Promoter methylation of RB1, P15, P16, and MGMT and their impact on the clinical course of pilocytic astrocytomas. Oncol Lett 15(2):1600–1606Google Scholar
- 43.Stewart LA (2002) Chemotherapy in adult high-grade glioma: a systematic review and meta-analysis of individual patient data from 12 randomised trials. Lancet Lond Engl 359(9311):1011–1018Google Scholar
- 44.Stupp R, Mason WP, van den Bent MJ et al (2005) Radiotherapy plus concomitant and adjuvant temozolomide for glioblastoma. N Engl J Med 352(10):987–996Google Scholar
- 45.Takeuchi K, Hoshino K (1977) Statistical analysis of factors affecting survival after glioblastoma multiforme. Acta Neurochir 37(1–2):57–73Google Scholar
- 46.Weber EL, Goebel EA (2005) Cerebral edema associated with Gliadel wafers: two case studies. Neuro-Oncol 7(1):84–89Google Scholar
- 47.Wemmert S, Bettscheider M, Alt S, Ketter R, Kammers K, Feiden W, Steudel W-I, Rahnenführer J, Urbschat S (2009) p15 promoter methylation—a novel prognostic marker in glioblastoma patients. Int J Oncol 34(6):1743–1748Google Scholar
- 48.Westphal M, Hilt DC, Bortey E, Delavault P, Olivares R, Warnke PC, Whittle IR, Jääskeläinen J, Ram Z (2003) A phase 3 trial of local chemotherapy with biodegradable carmustine (BCNU) wafers (Gliadel wafers) in patients with primary malignant glioma. Neuro-Oncol 5(2):79–88Google Scholar
- 49.Xu W, Yang H, Liu Y et al (2011) Oncometabolite 2-hydroxyglutarate is a competitive inhibitor of α-ketoglutarate-dependent dioxygenases. Cancer Cell 19(1):17–30Google Scholar
- 50.Yoshioka M, Matsutani T, Hara A, Hirono S, Hiwasa T, Takiguchi M, Iwadate Y (2018) Real-time methylation-specific PCR for the evaluation of methylation status of MGMT gene in glioblastoma. Oncotarget 9(45):27728–27735Google Scholar
- 51.Yu N, Yang Y, Li X, Zhang M, Huang J, Wang X, Long X (2016) MiR-26a inhibits proliferation and migration of HaCaT keratinocytes through regulating PTEN expression. Gene 594(1):117–124Google Scholar
- 52.Zhang Y-F, Zhang A-R, Zhang B-C, Rao Z-G, Gao J-F, Lv M-H, Wu Y-Y, Wang S-M, Wang R-Q, Fang D-C (2013) MiR-26a regulates cell cycle and anoikis of human esophageal adenocarcinoma cells through Rb1-E2F1 signaling pathway. Mol Biol Rep 40(2):1711–1720Google Scholar