Gliomas are the most common brain tumor type in children and adolescents. To date, diagnosis and therapy monitoring for these tumors rely on magnetic resonance imaging (MRI) and histopathological as well as molecular analyses of tumor tissue. Recently, liquid biopsies (LB) have emerged as promising tool for diagnosis and longitudinal tumor assessment potentially allowing for a more precise therapeutic management. However, the optimal strategy for monitoring gliomas by LB remains to be determined. In this study, we analyzed circulating tumor DNA (ctDNA) from 78 liquid biopsies (plasma n = 44, cerebrospinal fluid n = 34 (CSF)) of 35 glioma patients, determining H3F3A K28M (K27M) and BRAF V600E mutation allele frequency using droplet digital PCR (ddPCR). All results were correlated to clinically relevant parameters including diagnostic imaging and CSF aspiration site (ventricular vs lumbar) with respect to tumor localization. Regarding diagnostic accuracy, the calculated sensitivity score in the H3F3A K27M cohort was 84.61% for CSF and 73.68% for plasma. In the BRAF V600E cohort, we determined a sensitivity of 83.3% in plasma and 80% in CSF. The overall specificity was 100%. With respect to the CSF aspiration, the intra-operatively obtained CSF demonstrated 100% detection rate, followed by ventricular CSF obtained via Ommaya Reservoir/shunt puncture (93%) and CSF obtained via lumbar puncture (66%). Notably, this further correlated with the proximity of the CSF site to tumor localization. Longitudinal CSF monitoring demonstrated a good correlation to clinical and radiological disease evolution. Importantly, we show for the first time that monitoring BRAF V600E by ddPCR could serve as treatment response assessment in gliomas. In summary, our observation may inform recommendations with regard to location of CSF aspiration when incorporating LB into future treatment protocols.

Center for Cancer Research Medical University of Vienna Vienna Austria

Center for Precision Medicine University Hospital Brno Brno Czech Republic

Department of Biology Faculty of Medicine and Central European Institute of Technology Masaryk University Brno Czech Republic

Department of Biomedical Imaging and Image Guided Therapy Medical University of Vienna Vienna Austria

Department of Internal Medicine 1 Medical University of Vienna Vienna Austria

Department of Neurosurgery Medical University of Vienna Vienna Austria

Department of Pediatric Oncology University Hospital Brno and Faculty of Medicine Masaryk University Brno Czech Republic

Department of Pediatrics and Adolescent Medicine Comprehensive Center for Pediatrics and Comprehensive Cancer Center Medical University of Vienna Vienna Austria

Department of Radiation Oncology Medical University of Vienna Vienna Austria

Division of Neuropathology and Neurochemistry Department of Neurology Medical University of Vienna Vienna Austria

Research Center of Image Analysis and Artificial Intelligence Faculty of Medicine and Dentistry Danube Private University Krems Stein Austria

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Arthur C, Carlson LM, Svoboda J, Sandvik U, Jylhä C, Nordenskjöld M et al (2024) Liquid biopsy guides successful molecular targeted therapy of an inoperable pediatric brainstem neoplasm. NPJ Precis Oncol 8:44. 10.1038/s41698-024-00535-8 PubMed PMC

Azad TD, Bettegowda C (2022) Longitudinal monitoring of diffuse midline glioma using liquid biopsy. Neuro Oncol 24:1375–1376. 10.1093/neuonc/noac076 PubMed PMC

Bouffet E, Hansford JR, Garrè ML, Hara J, Plant-Fox A, Aerts I et al (2023) Dabrafenib plus Trametinib in Pediatric Glioma with BRAF V600 Mutations. N Engl J Med 389:1108–1120. 10.1056/NEJMoa2303815 PubMed

Buczkowicz P, Hoeman C, Rakopoulos P, Pajovic S, Letourneau L, Dzamba M et al (2014) Genomic analysis of diffuse intrinsic pontine gliomas identifies three molecular subgroups and recurrent activating ACVR1 mutations. Nat Genet 46:451–456. 10.1038/ng.2936 PubMed PMC

Cantor E, Wierzbicki K, Tarapore RS, Ravi K, Thomas C, Cartaxo R et al (2022) Serial H3K27M cell-free tumor DNA (cf-tDNA) tracking predicts ONC201 treatment response and progression in diffuse midline glioma. Neuro Oncol 24:1366–1374. 10.1093/neuonc/noac030 PubMed PMC

Capper D, Reifenberger G, French PJ, Schweizer L, Weller M, Touat M et al (2023) EANO guideline on rational molecular testing of gliomas, glioneuronal, and neuronal tumors in adults for targeted therapy selection. Neuro Oncol 25:813–826. 10.1093/neuonc/noad008 PubMed PMC

Castel D, Philippe C, Calmon R, Le Dret L, Truffaux N, Boddaert N et al (2015) Histone H3F3A and HIST1H3B K27M mutations define two subgroups of diffuse intrinsic pontine gliomas with different prognosis and phenotypes. Acta Neuropathol 130:815–827. 10.1007/s00401-015-1478-0 PubMed PMC

Christensen E, Birkenkamp-Demtröder K, Sethi H, Shchegrova S, Salari R, Nordentoft I et al (2019) Early detection of metastatic relapse and monitoring of therapeutic efficacy by ultra-deep sequencing of plasma cell-free dna in patients with urothelial bladder carcinoma. J Clin Oncol Off J Am Soc Clin Oncol 37:1547–1557. 10.1200/jco.18.02052 PubMed

Duffy MJ (2024) Circulating tumor DNA (ctDNA) as a biomarker for lung cancer: early detection, monitoring and therapy prediction. Tumour Biol J Int Soc Oncodev Biol Med 46:S283-s295. 10.3233/tub-220044 PubMed

Eibl RH, Schneemann M (2021) Liquid biopsy and primary brain tumors. Cancers (Basel). 10.3390/cancers13215429 PubMed PMC

Filbin MG, Tirosh I, Hovestadt V, Shaw ML, Escalante LE, Mathewson ND et al (2018) Developmental and oncogenic programs in H3K27M gliomas dissected by single-cell RNA-seq. Science 360:331–335. 10.1126/science.aao4750 PubMed PMC

Gojo J, Pavelka Z, Zapletalova D, Schmook MT, Mayr L, Madlener S et al (2019) Personalized treatment of H3K27M-mutant pediatric diffuse gliomas provides improved therapeutic opportunities. Front Oncol 9:1436. 10.3389/fonc.2019.01436 PubMed PMC

Hargrave DR, Terashima K, Hara J, Kordes UR, Upadhyaya SA, Sahm F et al (2023) Phase II trial of dabrafenib plus trametinib in relapsed/refractory BRAF V600–mutant pediatric high-grade glioma. J Clin Oncol 41:5174–5183. 10.1200/jco.23.00558 PubMed PMC

Hoffman LM, Van Veldhuijzen Zanten SEM, Colditz N, Baugh J, Chaney B, Hoffmann M et al (2018) Clinical, radiologic, pathologic, and molecular characteristics of long-term survivors of diffuse intrinsic pontine glioma (DIPG): a collaborative report from the International and European Society for Pediatric Oncology DIPG Registries. J Clin Oncol 36:1963–1972. 10.1200/jco.2017.75.9308 PubMed PMC

Jansen MH, van Veldhuijzen Zanten SE, Sanchez Aliaga E, Heymans MW, Warmuth-Metz M, Hargrave D et al (2015) Survival prediction model of children with diffuse intrinsic pontine glioma based on clinical and radiological criteria. Neuro Oncol 17:160–166. 10.1093/neuonc/nou104 PubMed PMC

Jones C, Baker SJ (2014) Unique genetic and epigenetic mechanisms driving paediatric diffuse high-grade glioma. Nat Rev Cancer 14:651–661. 10.1038/nrc3811 PubMed PMC

Jones C, Karajannis MA, Jones DTW, Kieran MW, Monje M, Baker SJ et al (2017) Pediatric high-grade glioma: biologically and clinically in need of new thinking. Neuro Oncol 19:153–161. 10.1093/neuonc/now101 PubMed PMC

Kang KM, Muralidharan K, Yekula A, Small JL, Rosh ZS, Jones PS et al (2021) Blood-based detection of BRAF V600E in gliomas and brain tumor metastasis. Cancers. 10.3390/cancers13061227 PubMed PMC

Khuong-Quang D-A, Buczkowicz P, Rakopoulos P, Liu X-Y, Fontebasso AM, Bouffet E et al (2012) K27M mutation in histone H3.3 defines clinically and biologically distinct subgroups of pediatric diffuse intrinsic pontine gliomas. Acta Neuropathol 124:439–447. 10.1007/s00401-012-0998-0 PubMed PMC

Krynina O, de Ståhl TD, Jylhä C, Arthur C, Giraud G, Nyman P et al (2024) The potential of liquid biopsy for detection of the KIAA1549-BRAF fusion in circulating tumor DNA from children with pilocytic astrocytoma. Neurooncol Adv. 10.1093/noajnl/vdae008 PubMed PMC

Kuligina E, Moiseyenko F, Belukhin S, Stepanova E, Zakharova M, Chernobrivtseva V et al (2021) Tumor irradiation may facilitate the detection of tumor-specific mutations in plasma. World J Clin Oncol 12:1215–1226. 10.5306/wjco.v12.i12.1215 PubMed PMC

Li JH, He ZQ, Lin FH, Chen ZH, Yang SY, Duan H et al (2019) Assessment of ctDNA in CSF may be a more rapid means of assessing surgical outcomes than plasma ctDNA in glioblastoma. Mol Cell Probes 46:101411. 10.1016/j.mcp.2019.06.001 PubMed

Louis DN, Perry A, Wesseling P, Brat DJ, Cree IA, Figarella-Branger D et al (2021) The 2021 WHO classification of tumors of the central nervous system: a summary. Neuro Oncol 23:1231–1251. 10.1093/neuonc/noab106 PubMed PMC

Mackay A, Burford A, Carvalho D, Izquierdo E, Fazal-Salom J, Taylor KR et al (2017) Integrated molecular meta-analysis of 1000 pediatric high-grade and diffuse intrinsic pontine glioma. Cancer Cell 32:520-537.e525. 10.1016/j.ccell.2017.08.017 PubMed PMC

Madlener S, Gojo J (2020) Liquid biomarkers for pediatric brain tumors: biological features, advantages and perspectives. J Pers Med. 10.3390/jpm10040254 PubMed PMC

Ostrom QT, Adel Fahmideh M, Cote DJ, Muskens IS, Schraw JM, Scheurer ME et al (2019) Risk factors for childhood and adult primary brain tumors. Neuro Oncol 21:1357–1375. 10.1093/neuonc/noz123 PubMed PMC

Pagès M, Beccaria K, Boddaert N, Saffroy R, Besnard A, Castel D et al (2018) Co-occurrence of histone H3 K27M and BRAF V600E mutations in paediatric midline grade I ganglioglioma. Brain Pathol 28:103–111. 10.1111/bpa.12473 PubMed PMC

Pagès M, Rotem D, Gydush G, Reed S, Rhoades J, Ha G et al (2022) Liquid biopsy detection of genomic alterations in pediatric brain tumors from cell-free DNA in peripheral blood, CSF, and urine. Neuro Oncol 24:1352–1363. 10.1093/neuonc/noab299 PubMed PMC

Panditharatna E, Kilburn LB, Aboian MS, Kambhampati M, Gordish-Dressman H, Magge SN et al (2018) Clinically relevant and minimally invasive tumor surveillance of pediatric diffuse midline gliomas using patient-derived liquid biopsy. Clin Cancer Res Off J Am Assoc Cancer Res 24:5850–5859. 10.1158/1078-0432.Ccr-18-1345 PubMed PMC

Ryall S, Krishnatry R, Arnoldo A, Buczkowicz P, Mistry M, Siddaway R et al (2016) Targeted detection of genetic alterations reveal the prognostic impact of H3K27M and MAPK pathway aberrations in paediatric thalamic glioma. Acta Neuropathol Commun 4:93. 10.1186/s40478-016-0353-0 PubMed PMC

Schwartzentruber J, Korshunov A, Liu XY, Jones DT, Pfaff E, Jacob K et al (2012) Driver mutations in histone H3.3 and chromatin remodelling genes in paediatric glioblastoma. Nature 482:226–231. 10.1038/nature10833 PubMed

Soffietti R, Bettegowda C, Mellinghoff IK, Warren KE, Ahluwalia MS, De Groot JF et al (2022) Liquid biopsy in gliomas: a RANO review and proposals for clinical applications. Neuro Oncol 24:855–871. 10.1093/neuonc/noac004 PubMed PMC

Stepien N, Senfter D, Furtner J, Haberler C, Dorfer C, Czech T et al (2023) Proof-of-concept for liquid biopsy disease monitoring of myc-amplified group 3 medulloblastoma by droplet digital PCR. Cancers. 10.3390/cancers15092525 PubMed PMC

Tie J, Cohen JD, Lahouel K, Lo SN, Wang Y, Kosmider S et al (2022) Circulating tumor dna analysis guiding adjuvant therapy in stage II colon cancer. N Engl J Med 386:2261–2272. 10.1056/NEJMoa2200075 PubMed PMC

Tie J, Cohen JD, Wang Y, Christie M, Simons K, Lee M et al (2019) Circulating tumor DNA analyses as markers of recurrence risk and benefit of adjuvant therapy for stage III colon cancer. JAMA Oncol 5:1710–1717. 10.1001/jamaoncol.2019.3616 PubMed PMC

Traylor JI, Kuo JS (2019) “Liquid biopsy” of circulating tumor DNA characterizes treatment response in pediatric patients diffuse midline glioma. Neurosurgery 85:E167-e168. 10.1093/neuros/nyz029 PubMed

Trevethan R (2017) Sensitivity, specificity, and predictive values: foundations, pliabilities, and pitfalls in research and practice. Front Public Health 5:307. 10.3389/fpubh.2017.00307 PubMed PMC

Venneti S, Kawakibi AR, Ji S, Waszak SM, Sweha SR, Mota M et al (2023) Clinical efficacy of ONC201 in H3K27M-mutant diffuse midline gliomas is driven by disruption of integrated metabolic and epigenetic pathways. Cancer Discov 13:2370–2393. 10.1158/2159-8290.Cd-23-0131 PubMed PMC

Wierzbicki K, Ravi K, Franson A, Bruzek A, Cantor E, Harris M et al (2020) Targeting and therapeutic monitoring of H3K27M-mutant glioma. Curr Oncol Rep 22:19. 10.1007/s11912-020-0877-0 PubMed PMC

Yekula A, Muralidharan K, Rosh ZS, Youngkin AE, Kang KM, Balaj L et al (2020) Liquid biopsy strategies to distinguish progression from pseudoprogression and radiation necrosis in glioblastomas. Adv Biosyst 4:e2000029. 10.1002/adbi.202000029 PubMed PMC