{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,5]],"date-time":"2026-10-05T21:50:56Z","timestamp":1791237056043,"version":"4.1.0"},"reference-count":43,"publisher":"Radiation Research Society","issue":"2","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Radiation Research"],"published-print":{"date-parts":[[2026,5,26]]},"abstract":"<jats:p>Glioblastoma WHO CNS Grade 4 (GBM) is associated with poor prognosis and high recurrence rates despite the current therapeutic interventions. Conventional imaging bears restrictions in detecting tumor recurrence and discriminating it from tissue necrosis and post-operative scar tissues. In a previous study, we conducted a whole-transcriptome screening using next-generation sequencing (NGS) on whole blood samples (n = 33) from seven patients and tumor biopsies to identify tumor-specific gene expression patterns. This revealed downregulation post-surgery, with a return to pre-surgery baseline levels at the time of tumor recurrence. In the screening phase, we were able to identify 374 genes across all patients, and in this study, we aimed to validate the most predictive genes identified in the screening phase using the same samples with qRT-PCR (customized TaqMan array cards). A subset of candidate genes was selected for each patient to reduce the number of screening genes to be validated, resulting in a total of 94 genes for validation using qRT-PCR. All genes were measured in all samples, thus allowing a search for genes methodologically confirming the NGS candidate genes as well as examining qRT-PCR-based gene expression patterns, which were not captured by NGS in certain patients. Filter criteria included a post-surgery fold change (FC) \u2264 0.6 across all or most time points and already cited GBM-related genes. From whole blood, total RNA was isolated, converted into cDNA, and 6,270 gene expression measurements were performed employing custom TaqMan array cards (qRT-PCR). We were able to successfully validate NGS candidate genes for three patients out of seven. Differential gene expression (DGE) of both methods was comparable for these patients (e.g., r2 = 0.76, P &lt; 0.001 for patient #1). Between 38\u201362% of NGS candidate genes per patient could be successfully validated, but only in patients where the screening comprised \u226520 NGS candidate genes. QRT-PCR also detected tumor recurrence-associated expression patterns in genes that were not captured by NGS, but 2\u20139-fold more candidates were observed for NGS-selected genes. Furthermore, in a patient receiving additional chemotherapy (CTx) during the tumor recurrence phase, a reduction in DGE of 13 genes was observed. In another patient, DGE indicated recurrence 13 days before radiological examinations confirmed it. The evidence, based on screening and validated, suggests a potential association, but a coincidental link cannot be ruled out. Our findings may point to the possible utility of gene expression measurements in peripheral whole blood as a simplified liquid biopsy method for GBM tumor recurrence. However, further prospective cohort studies with improved control of sampling during clinical follow-up and radiological examinations are needed to strengthen the presumed causality and the clinical value of early gene expression changes as an indicator of tumor recurrence.<\/jats:p>","DOI":"10.1667\/rade-25-00226.1","type":"journal-article","created":{"date-parts":[[2026,5,27]],"date-time":"2026-05-27T00:03:01Z","timestamp":1779840181000},"page":"197-206","source":"Crossref","is-referenced-by-count":0,"title":["Using Gene Expression Changes for Monitoring Glioblastoma WHO CNS Grade 4 Recurrence: Validation of NGS Candidate Genes via qRT-PCR"],"prefix":"10.1667","volume":"206","author":[{"given":"R.","family":"Muhtadi","sequence":"first","affiliation":[{"name":"Bundeswehr Institute of Radiobiology, Munich, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"N.","family":"Schmid","sequence":"additional","affiliation":[{"name":"Bundeswehr Institute of Radiobiology, Munich, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"D.","family":"Bernhardt","sequence":"additional","affiliation":[{"name":"Department of Radiation Oncology, TUM School of Medicine and Health, Munich, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"G.","family":"Multhoff","sequence":"additional","affiliation":[{"name":"Department of Radiation Oncology, TUM School of Medicine and Health, Munich, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"L.","family":"H\u00f6nikl","sequence":"additional","affiliation":[{"name":"Department of Neurosurgery, TUM School of Medicine, Klinikum Rechts der Isar, Munich, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"S. E.","family":"Combs","sequence":"additional","affiliation":[{"name":"Department of Radiation Oncology, TUM School of Medicine and Health, Munich, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"S. M.","family":"Krieg","sequence":"additional","affiliation":[{"name":"Department of Neurosurgery, TUM School of Medicine, Klinikum Rechts der Isar, Munich, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"J.","family":"Gempt","sequence":"additional","affiliation":[{"name":"Department of Neurosurgery, TUM School of Medicine, Klinikum Rechts der Isar, Munich, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"M.","family":"Bernhard","sequence":"additional","affiliation":[{"name":"Department of Neurosurgery, TUM School of Medicine, Klinikum Rechts der Isar, Munich, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"V.","family":"Barsegian","sequence":"additional","affiliation":[{"name":"Institute of Nuclear Medicine, Helios Kliniken, Schwerin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"M.","family":"Lindemann","sequence":"additional","affiliation":[{"name":"Institute for Transfusion Medicine, University Hospital, Essen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"M.","family":"Kasper","sequence":"additional","affiliation":[{"name":"Bundeswehr Institute of Radiobiology, Munich, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"S.","family":"Stewart","sequence":"additional","affiliation":[{"name":"Bundeswehr Institute of Radiobiology, Munich, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"M.","family":"Port","sequence":"additional","affiliation":[{"name":"Bundeswehr Institute of Radiobiology, Munich, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"S.","family":"Eder","sequence":"additional","affiliation":[{"name":"Bundeswehr Institute of Radiobiology, Munich, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"C. D.","family":"Diehl","sequence":"additional","affiliation":[{"name":"Department of Radiation Oncology, TUM School of Medicine and Health, Munich, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"P.","family":"Ostheim","sequence":"additional","affiliation":[{"name":"Bundeswehr Institute of Radiobiology, Munich, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"M.","family":"Abend","sequence":"additional","affiliation":[{"name":"Bundeswehr Institute of Radiobiology, Munich, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1243","reference":[{"key":"1","doi-asserted-by":"publisher","unstructured":"RH Eibl  and M Schneemann  \"Explor Target Antitumor Ther.\"  4, \"Liquid biopsy and glioblastoma.\" 28 (2023)","DOI":"10.37349\/etat.2023.00121"},{"key":"2","doi-asserted-by":"publisher","unstructured":"H Ali, R Harting, R de Vries, et al.  \"Front Oncol.\"  11, \"Blood-based biomarkers for glioma in the context of gliomagenesis: A systematic review.\" 665235 (2021)","DOI":"10.3389\/fonc.2021.665235"},{"key":"3","doi-asserted-by":"publisher","unstructured":"R Stupp, W Mason, M Bent, et al.  \"N Engl J Med.\"  352, \"Radiotherapy plus concomitant and adjuvant temozolomide for glioblastoma.\" 987 (2005)","DOI":"10.1056\/NEJMoa043330"},{"key":"4","doi-asserted-by":"publisher","unstructured":"SA Grossman, X Ye, S Piantadosi, et al.  \"Clin Cancer Res.\"  16, \"Survival of patients with newly diagnosed glioblastoma treated with radiation and temozolomide in research studies in the United States.\" 2443 (2010)","DOI":"10.1158\/1078-0432.CCR-09-3106"},{"key":"5","unstructured":"AF Tamimi  and M Juweid  \"Glioblastoma.\"  \"Epidemiology and Outcome of Glioblastoma.\" (2017)"},{"key":"6","doi-asserted-by":"publisher","unstructured":"B Campos, LR Olsen, T Urup, et al.  \"Oncogene.\"  35, \"A comprehensive profile of recurrent glioblastoma.\" 5819 (2016)","DOI":"10.1038\/onc.2016.85"},{"key":"7","doi-asserted-by":"publisher","unstructured":"A Saenz-Anto\u00f1anzas, J Auzmendi-Iriarte, E Carrasco-Garcia, et al.  \"Cancers.\"  11, \"Liquid biopsy in glioblastoma: Opportunities, applications and challenges.\" 950 (2019)","DOI":"10.3390\/cancers11070950"},{"key":"8","doi-asserted-by":"publisher","unstructured":"M Martucci, R Russo, C Giordano, et al.  \"Cancers.\"  15, \"Advanced magnetic resonance imaging in the evaluation of treated glioblastoma: A pictorial essay.\" (2023)","DOI":"10.3390\/cancers15153790"},{"key":"9","doi-asserted-by":"publisher","unstructured":"OL Chinot, DR Macdonald, LE Abrey, et al.  \"Curr Neurol Neurosci Rep.\"  13, \"Response assessment criteria for glioblastoma: practical adaptation and implementation in clinical trials of antiangiogenic therapy.\" 347 (2013)","DOI":"10.1007\/s11910-013-0347-2"},{"key":"10","doi-asserted-by":"publisher","unstructured":"N Verma, MC Cowperthwaite, MG Burnett, et al.  \"Neuro-oncology.\"  15, \"Differentiating tumor recurrence from treatment necrosis: a review of neuro-oncologic imaging strategies.\" 515 (2013)","DOI":"10.1093\/neuonc\/nos307"},{"key":"11","doi-asserted-by":"publisher","unstructured":"G Dwarshuis, LL Kroon, D Brandsma, et al.  \"Acta Neuropathologica.\"  149, \"Liquid biopsies for the monitoring of gliomas and brain metastases in adults.\" 37 (2025)","DOI":"10.1007\/s00401-025-02880-9"},{"key":"12","doi-asserted-by":"publisher","unstructured":"BP de Lima, LS Ferraz, S Devalle, et al.  \"Biomolecules.\"  15, \"Liquid biopsy-derived tumor biomarkers for clinical applications in glioblastoma.\" 658 (2025)","DOI":"10.3390\/biom15050658"},{"key":"13","doi-asserted-by":"publisher","unstructured":"K Anitha, B Posinasetty, K Naveen Kumari, et al.  \"Clinica Chimica Acta.\"  554, \"Liquid biopsy for precision diagnostics and therapeutics.\" 117746 (2024)","DOI":"10.1016\/j.cca.2023.117746"},{"key":"14","doi-asserted-by":"publisher","unstructured":"A Ray  and TK Vohra  \"Neurooncol Advances.\"  4, \"Liquid biopsy\u2014from bench to bedside.\" ii66 (2022)","DOI":"10.1093\/noajnl\/vdac037"},{"key":"15","unstructured":"F Xiao, S Lv, Z Zong, et al.  \"Am J Transl Res.\"  12, \"Cerebrospinal fluid biomarkers for brain tumor detection: clinical roles and current progress.\" 1379 (20)"},{"key":"16","doi-asserted-by":"publisher","unstructured":"JCM Wan, C Massie, J Garcia-Corbacho, et al.  \"Nat Rev Cancer.\"  17, \"Liquid biopsies come of age: towards implementation of circulating tumour DNA.\" 223 (2017)","DOI":"10.1038\/nrc.2017.7"},{"key":"17","doi-asserted-by":"publisher","unstructured":"LG. Tataranu,  \"Medicina.\"  61, \"Liquid biopsy as a diagnostic and monitoring tool in glioblastoma.\" 716 (2025)","DOI":"10.3390\/medicina61040716"},{"key":"18","doi-asserted-by":"publisher","unstructured":"L Dai, Z Liu, Y Zhu, et al.  \"Heliyon.\"  9, \"Genome-wide methylation analysis of circulating tumor DNA: A new biomarker for recurrent glioblastom.\" (2023)","DOI":"10.1016\/j.heliyon.2023.e14339"},{"key":"19","doi-asserted-by":"publisher","unstructured":"D Shyr  and Q Liu  \"Biol Proced Online.\"  15, \"Next generation sequencing in cancer research and clinical application.\" 4 (2013)","DOI":"10.1186\/1480-9222-15-4"},{"key":"20","doi-asserted-by":"publisher","unstructured":"M Kubista, JM Andrade, M Bengtsson, et al.  \"Mol Aspects Med.\"  27, \"The real-time polymerase chain reaction.\" 95 (2006)","DOI":"10.1016\/j.mam.2005.12.007"},{"key":"21","doi-asserted-by":"publisher","unstructured":"R Muhtadi, D Bernhardt, G Multhoff, et al.  \"Cancers.\"  16, \"Liquid biopsy in whole blood for identification of gene expression patterns (mRNA and miRNA) associated with recurrence of glioblastoma WHO CNS grade 4.\" (2024)","DOI":"10.3390\/cancers16132345"},{"key":"22","doi-asserted-by":"publisher","unstructured":"A Bardelli  and K Pantel  \"Cancer Cell.\"  31, \"Liquid biopsies, what we do not know (yet).\" 172 (2017)","DOI":"10.1016\/j.ccell.2017.01.002"},{"key":"24","doi-asserted-by":"publisher","unstructured":"M Port, F H\u00e9rodin, M Valente, et al.  \"Ann Hematol.\"  96, \"Gene expression signature for early prediction of late occurring pancytopenia in irradiated baboons.\" 859 (2017)","DOI":"10.1007\/s00277-017-2952-7"},{"key":"25","unstructured":"\"NCBI Gene Database (cited 2025 June 6, 2025).\""},{"key":"26","doi-asserted-by":"publisher","unstructured":"MA Dymova, EV Kuligina, VA Richter,  \"Int J Mol Sci.\"  22, \"Molecular mechanisms of drug resistance in glioblastoma.\" (2021)","DOI":"10.3390\/ijms22126385"},{"key":"27","doi-asserted-by":"publisher","unstructured":"G P\u00f6pperl, C G\u00f6tz, W Rachinger, et al.  \"Eur J Nucl Med Mol Imaging.\"  31, \"Value of O-(2-(18F)fluoroethyl)-l-tyrosine PET for the diagnosis of recurrent glioma.\" 1464 (2004)","DOI":"10.1007\/s00259-004-1590-1"},{"key":"28","doi-asserted-by":"crossref","unstructured":"P Ostheim, M Majewski, Z Gluzman-Poltorak, et al.  \"Radiat Res.\"  195, \"Predicting the radiation sensitivity of male and female rhesus macaques using gene expression.\" 25 (2020)","DOI":"10.1667\/RADE-20-00161.1"},{"key":"29","doi-asserted-by":"publisher","unstructured":"M Majewski, P Ostheim, Z Gluzman-Poltorak, et al.  \"PLOS ONE.\"  16, \"Gene expression changes in male and female rhesus macaque 60 days after irradiation.\" e0254344 (2021)","DOI":"10.1371\/journal.pone.0254344"},{"key":"30","doi-asserted-by":"publisher","unstructured":"P Ostheim, M Miederer, M Schreckenberger, et al.  \"Int J Radiat Biol.\"  98, \"mRNA and small RNA gene expression changes in peripheral blood to detect internal Ra-223 exposure.\" 900 (2022)","DOI":"10.1080\/09553002.2021.1998705"},{"key":"31","doi-asserted-by":"publisher","unstructured":"D Enderle, A Spiel, CM Coticchia, et al.  \"PLoS One.\"  10, \"Characterization of RNA from exosomes and other extracellular vesicles isolated by a novel spin column-based method.\" e0136133 (2015)","DOI":"10.1371\/journal.pone.0136133"},{"key":"32","doi-asserted-by":"publisher","unstructured":"MA Aziz, B Seo, HM Hussaini, et al.  \"J Nucleic Acids.\"  2022, \"Comparing two methods for the isolation of exosomes.\" 8648373 (2022)","DOI":"10.1155\/2022\/8648373"},{"key":"33","doi-asserted-by":"publisher","unstructured":"M Kenzelmann, R Klaren, M Hergenhahn, et al.  \"Genomics.\"  83, \"High-accuracy amplification of nanogram total RNA amounts for gene profiling.\" 550 (2004)","DOI":"10.1016\/j.ygeno.2003.09.026"},{"key":"35","doi-asserted-by":"publisher","unstructured":"A Yekula, T Hsia, RR Kitchen, et al.  \"Neurooncol Adv.\"  5, \"Longitudinal analysis of serum-derived extracellular vesicle RNA to monitor dacomitinib treatment response in EGFR-amplified recurrent glioblastoma patients.\" vdad104 (2023)","DOI":"10.1093\/noajnl\/vdad104"},{"key":"36","doi-asserted-by":"publisher","unstructured":"M Niyazi, A Pitea, M Mittelbronn, et al.  \"Oncotarget.\"  7, \"A 4-miRNA signature predicts the therapeutic outcome of glioblastoma.\" 45764 (2016)","DOI":"10.18632\/oncotarget.9945"},{"key":"37","doi-asserted-by":"publisher","unstructured":"D Qi, Y Geng, J Cardenas, et al.  \"NPJ Genom Med.\"  8, \"Transcriptomic analyses of patient peripheral blood with hemoglobin depletion reveal glioblastoma biomarkers.\" 2 (2023)","DOI":"10.1038\/s41525-022-00348-3"},{"key":"38","doi-asserted-by":"publisher","unstructured":"T Wu, Y Li, B Liu, et al.  \"PLoS One.\"  11, \"Expression of ferritin light chain (FTL) Is elevated in glioblastoma, and FTL silencing inhibits glioblastoma cell proliferation via the GADD45\/JNK pathway.\" e0149361 (2016)","DOI":"10.1371\/journal.pone.0149361"},{"key":"39","doi-asserted-by":"publisher","unstructured":"ZW Hu, L Chen, RQ Ma, et al.  \"Aging (Albany NY).\"  13, \"Comprehensive analysis of ferritin subunits expression and positive correlations with tumor-associated macrophages and T regulatory cells infiltration in most solid tumors.\" 11491 (2021)","DOI":"10.18632\/aging.202841"},{"key":"40","doi-asserted-by":"publisher","unstructured":"V Ravi, AB Madhankumar, T Abraham, et al.  \"PLoS One.\"  14, \"Liposomal delivery of ferritin heavy chain 1 (FTH1) siRNA in patient xenograft derived glioblastoma initiating cells suggests different sensitivities to radiation and distinct survival mechanisms.\" e0221952 (2019)","DOI":"10.1371\/journal.pone.0221952"},{"key":"41","doi-asserted-by":"publisher","unstructured":"MY Alshahrani, EF Oghenemaro, J Rizaev, et al.  \"Human Immunol.\"  86, \"Exploring the modulation of TLR4 and its associated ncRNAs in cancer immunopathogenesis, with an emphasis on the therapeutic implications and mechanisms underlying drug resistance.\" 111188 (2025)","DOI":"10.1016\/j.humimm.2024.111188"},{"key":"42","doi-asserted-by":"publisher","unstructured":"C Zhu, X Chen, G Guan, et al.  \"Onco Targets Ther.\"  13, \"IFI30 Is a novel immune-related target with predicting value of prognosis and treatment response in glioblastoma.\" 1129 (2020)","DOI":"10.2147\/ott.S237162"},{"key":"43","doi-asserted-by":"publisher","unstructured":"Y Baba, D Nomoto, K Okadome, et al.  \"Cancer Sci.\"  111, \"Tumor immune microenvironment and immune checkpoint inhibitors in esophageal squamous cell carcinoma.\" 3132 (2020)","DOI":"10.1111\/cas.14541"},{"key":"23","doi-asserted-by":"crossref","unstructured":"M Port, F Herodin, M Valente, et al.  \"Radiat Res.\"  186, \"First generation gene expression signature for early prediction of late occurring hematological acute radiation syndrome in baboons.\" 39 (2016)","DOI":"10.1667\/RR14318.1"},{"key":"34","doi-asserted-by":"crossref","unstructured":"C Gardiner, D Di Vizio, S Sahoo, et al.  \"J Extracell Vesicles.\"  5, \"Techniques used for the isolation and characterization of extracellular vesicles: results of a worldwide survey.\" 32945 (2016)","DOI":"10.3402\/jev.v5.32945"}],"container-title":["Radiation Research"],"original-title":[],"link":[{"URL":"https:\/\/radiation-research.kglmeridian.com\/view\/journals\/rare\/aop\/article-10.1667-RADE-25-00226.1\/article-10.1667-RADE-25-00226.1.xml","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/radiation-research.kglmeridian.com\/downloadpdf\/journals\/rare\/aop\/article-10.1667-RADE-25-00226.1\/article-10.1667-RADE-25-00226.1.xml","content-type":"text\/html","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/radiation-research.kglmeridian.com\/downloadpdf\/journals\/rare\/aop\/article-10.1667-RADE-25-00226.1\/article-10.1667-RADE-25-00226.1.xml","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,10,5]],"date-time":"2026-10-05T20:52:49Z","timestamp":1791233569000},"score":1,"resource":{"primary":{"URL":"https:\/\/bioone.org\/journals\/radiation-research\/volume-206\/issue-2\/RADE-25-00226.1\/Using-Gene-Expression-Changes-for-Monitoring-Glioblastoma-WHO-CNS-Grade\/10.1667\/RADE-25-00226.1.full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,5,26]]},"references-count":43,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2026,8,1]]}},"URL":"https:\/\/doi.org\/10.1667\/rade-25-00226.1","relation":{},"ISSN":["0033-7587","1938-5404"],"issn-type":[{"value":"0033-7587","type":"print"},{"value":"1938-5404","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,5,26]]}}}