Low Detection Rate of H3K27M Mutations in Cerebrospinal Fluid Obtained from Lumbar Puncture in Newly Diagnosed Diffuse Midline Gliomas
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Updated: Nov 7, 2025
Detection and Monitoring of Tumor Associated Circulating DNA in Patient Biofluids
Published on: June 8, 2019
Jotaro On1, Manabu Natsumeda1, Jun Watanabe1
1Department of Neurosurgery, Brain Research Institute, Niigata University, Niigata 951-8585, Japan.
Abstract:
Recent studies have suggested the feasibility of detecting H3K27M mutations in the cerebrospinal fluid of diffuse midline glioma (DMG) patients. However, cerebrospinal fluid from patients in these studies were collected mainly during biopsy, ventriculo-peritoneal shunt procedures or postmortem. We assessed circulating tumor DNA (ctDNA) extracted from cerebrospinal fluid (CSF) and plasma in a series of 12 radiographically suspected and/or pathologically confirmed diffuse midline glioma patients and assessed for H3F3A K27M mutation using digital droplet PCR. In 10 patients, CSF was obtained by lumbar puncture at presentation. A definitive detection of H3F3A K27M mutation was achieved in only one case (10%); H3F3A K27M mutation was suspected in three other cases (30%). H3F3A K27M mutation was detected in two patients in CSF obtained by ventricular tap during a ventriculo-peritoneal shunt for obstructive hydrocephalus. Cases in which a definitive assessment was possible (definite H3F3A K27M or definite H3F3A wildtype) tended to be younger (median 7.5 years vs. 40.5 years; p = 0.07) and have a higher concentration of CSF protein (median 123 mg/dL vs. 27.5 mg/dL; p = 0.21) compared to nondefinite cases. Low proliferation and apoptotic rates seemed to be characteristics of DMG unfavorable for liquid biopsy. More advanced lesions with necrosis and evidence of dissemination were unlikely to be candidates for lumbar puncture due to the fear of exacerbating obstructive hydrocephalus. Methods to safely sample CSF and a more sensitive detection of ctDNA are necessary for reliable liquid biopsy of DMG at presentation.
Insights
Detecting H3K27M mutations in diffuse midline glioma (DMG) patients via liquid biopsy showed low sensitivity. Cerebrospinal fluid (CSF) analysis using digital droplet PCR was challenging, necessitating improved ctDNA detection methods for DMG diagnosis.
Area of Science:
- Neuro-oncology
- Molecular Diagnostics
- Genetics
Background:
- Diffuse midline glioma (DMG) is an aggressive brain tumor often characterized by H3K27M mutations.
- Previous studies suggested cerebrospinal fluid (CSF) analysis for H3K27M mutation detection in DMG, but sample collection methods were limited.
- Liquid biopsy offers a less invasive approach for molecular profiling in neuro-oncology.
Purpose of the Study:
- To assess the feasibility of detecting H3F3A K27M mutations in CSF and plasma of DMG patients using circulating tumor DNA (ctDNA).
- To evaluate the efficacy of digital droplet PCR for H3K27M mutation detection in liquid biopsies from DMG patients.
- To identify factors influencing the success of liquid biopsy for H3K27M mutation detection in DMG.
Main Methods:
- Analyzed CSF and plasma samples from 12 DMG patients for H3F3A K27M mutation using digital droplet PCR.
- Collected CSF via lumbar puncture at presentation in 10 patients.
- Collected CSF during ventriculo-peritoneal shunt procedures in 2 patients.
Main Results:
- Definitive H3F3A K27M mutation detection was achieved in only 10% (1/12) of patients via CSF.
- H3F3A K27M mutation was suspected in 30% (3/12) of patients.
- Mutation detection was higher in CSF obtained during shunt procedures (2/2) compared to lumbar puncture (1/10).
- Younger age and higher CSF protein concentration were associated with definitive mutation assessment.
- Low tumor proliferation and apoptotic rates were unfavorable for liquid biopsy.
- Advanced lesions with necrosis or dissemination limited lumbar puncture feasibility.
Conclusions:
- Current ctDNA detection methods and CSF sampling techniques limit the reliability of liquid biopsy for H3K27M mutation detection in DMG at presentation.
- Improved CSF sampling strategies and more sensitive ctDNA detection assays are crucial for advancing liquid biopsy in DMG management.
- Further research is needed to optimize liquid biopsy protocols for accurate and timely diagnosis of DMG.
