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Updated: Jun 8, 2025
Live-3D-Cell Immunocytochemistry Assays of Pediatric Diffuse Midline Glioma
Published on: November 11, 2021
Extracellular Histones Profiles of Pediatric H3K27-Altered Diffuse Midline Glioma
Diana Buzova1, Lucia Lisa Petrilli2, Jan Frohlich3
1Department of Adaptive Biotechnologies, Global Change Research Institute CAS, Brno, Czech Republic.
Background:
Diffuse midline glioma, H3 K27-altered (DMG) is a fatal tumour that arises in the midline structures of the brain. When located in the pons, it is more commonly referred to as diffuse intrinsic pontine glioma (DIPG). DMG/DIPG is usually diagnosed when children are < 10 years, and it has a median overall survival of < 12 months after diagnosis. Radiological imaging is still the gold standard for DIPG diagnosis while the use of biopsy procedures led to our knowledge on its biology, such as with the identification of the canonical histone H3K27M mutation. However, the need to improve survival encourages the development of non-invasive, fast and inexpensive assays on biofluids for optimizing molecular diagnoses in DMG/DIPG. Here, we propose a rapid, new, imaging and epigenetics-based approach to diagnose DMG/DIPG in the plasma of paediatric patients.
Methods:
A total of 20 healthy children (mean age: 10.5 years) and 24 children diagnosed with DMG/DIPG (mean age: 8.5 years) were recruited. Individual histones (H2A, H2B, H3, H4, macroH2A1.1 and macroH2A1.2), histone dimers and nucleosomes were assayed in biofluids by means of a new advanced flow cytometry ImageStream(X)-adapted method.
Results:
We report a significant increase in circulating histone dimers and tetramers (macroH2A1.1/H2B versus control: p value < 0.0001; macroH2A1.2/H2B versus control: p value < 0.0001; H2A/H2B versus control: p value < 0.0001; H3/H4 versus control: p value = 0.008; H2A/H2B/H3/H4 versus control: p value < 0.0001) and a significant downregulation of individual histones (H2B versus control: p value < 0.0001; H3 versus control: p value < 0.0001; H4 versus control: p value < 0.0001). Moreover, histones were also detectable in the cerebrospinal fluid (CSF) of patients with DMG/DIPG and in the supernatant of SF8628, OPBG-DIPG002 and OPBG-DIPG004 DMG/DIPG cell lines, with patterns mostly similar to each other, but distinct compared to blood plasma.
Conclusions:
In summary, we identified circulating histone signatures able to detect the presence of DMG/DIPG in biofluids of children, using a rapid and non-invasive ImageStream(X)-based imaging technology, which may improve diagnosis and benefit the patients.
Insights
Researchers developed a new blood test to detect diffuse midline glioma, H3 K27-altered (DMG) in children. This non-invasive method identifies specific circulating histone patterns, potentially improving early diagnosis and patient outcomes for this rare brain tumor.
Area of Science:
- Oncology
- Molecular Biology
- Biochemistry
Background:
- Diffuse midline glioma, H3 K27-altered (DMG), including diffuse intrinsic pontine glioma (DIPG), is a pediatric brain tumor with a poor prognosis.
- Current diagnosis relies on radiological imaging, with biopsies providing biological insights but posing risks.
- There is a critical need for non-invasive, rapid, and cost-effective diagnostic methods for DMG/DIPG.
Purpose of the Study:
- To develop and validate a novel, non-invasive assay for the molecular diagnosis of DMG/DIPG in pediatric patients.
- To identify specific circulating histone signatures in biofluids indicative of DMG/DIPG.
- To utilize advanced imaging flow cytometry for rapid and accurate detection.
Main Methods:
- Recruited 20 healthy children and 24 children diagnosed with DMG/DIPG.
- Assayed individual histones, histone dimers, and nucleosomes in biofluids using an ImageStream(X)-adapted flow cytometry method.
- Analyzed plasma, cerebrospinal fluid (CSF), and cell line supernatants for histone patterns.
Main Results:
- Observed a significant increase in circulating histone dimers and tetramers in DMG/DIPG patients compared to controls.
- Detected a significant downregulation of individual histones (H2B, H3, H4) in DMG/DIPG patients.
- Histone patterns in CSF and cell line supernatants showed similarities to each other but differed from plasma patterns.
Conclusions:
- Identified specific circulating histone signatures for the detection of DMG/DIPG in pediatric biofluids.
- The developed ImageStream(X)-based imaging technology offers a rapid and non-invasive approach for DMG/DIPG diagnosis.
- This new method holds potential to improve diagnostic accuracy and benefit patients with DMG/DIPG.
