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Updated: Apr 18, 2026
Live-3D-Cell Immunocytochemistry Assays of Pediatric Diffuse Midline Glioma
Published on: November 11, 2021
Cell migration in paediatric glioma; characterisation and potential therapeutic targeting
J V Cockle1, S Picton2, J Levesley3
11] Leeds Institute of Cancer Studies and Pathology, University of Leeds, Wellcome Trust Brenner Building, St James's University Hospital, Leeds, LS9 7TF, UK [2] Yorkshire Regional Centre for Paediatric Oncology and Haematology, Leeds General Infirmary, Great George Street, Leeds, LS1 3EX, UK.
Background:
Paediatric high grade glioma (pHGG) and diffuse intrinsic pontine glioma (DIPG) are highly aggressive brain tumours. Their invasive phenotype contributes to their limited therapeutic response, and novel treatments that block brain tumour invasion are needed.
Methods:
Here, we examine the migratory characteristics and treatment effect of small molecule glycogen synthase kinase-3 inhibitors, lithium chloride (LiCl) and the indirubin derivative 6-bromoindirubin-oxime (BIO), previously shown to inhibit the migration of adult glioma cells, on two pHGG cell lines (SF188 and KNS42) and one patient-derived DIPG line (HSJD-DIPG-007) using 2D (transwell membrane, immunofluorescence, live cell imaging) and 3D (migration on nanofibre plates and spheroid invasion in collagen) assays.
Results:
All lines were migratory, but there were differences in morphology and migration rates. Both LiCl and BIO reduced migration and instigated cytoskeletal rearrangement of stress fibres and focal adhesions when viewed by immunofluorescence. In the presence of drugs, loss of polarity and differences in cellular movement were observed by live cell imaging.
Conclusions:
Ours is the first study to demonstrate that it is possible to pharmacologically target migration of paediatric glioma in vitro using LiCl and BIO, and we conclude that these agents and their derivatives warrant further preclinical investigation as potential anti-migratory therapeutics for these devastating tumours.
Insights
Small molecules lithium chloride (LiCl) and 6-bromoindirubin-oxime (BIO) effectively inhibited paediatric glioma cell migration in vitro. These findings suggest potential new anti-migratory therapies for aggressive paediatric brain tumours.
Area of Science:
- Neuro-oncology
- Cellular Biology
- Pharmacology
Background:
- Paediatric high-grade glioma (pHGG) and diffuse intrinsic pontine glioma (DIPG) are aggressive brain tumours.
- Tumour invasion limits therapeutic efficacy, necessitating novel anti-invasion strategies.
- Targeting glioma cell migration is a critical unmet need.
Purpose of the Study:
- To investigate the efficacy of glycogen synthase kinase-3 inhibitors (LiCl and BIO) on paediatric glioma cell migration.
- To assess the impact of these inhibitors on cellular morphology and migratory behaviour.
- To evaluate the potential of these compounds as anti-migratory therapeutics for paediatric brain tumours.
Main Methods:
- Utilized two pHGG cell lines (SF188, KNS42) and one DIPG line (HSJD-DIPG-007).
- Employed 2D assays (transwell, immunofluorescence, live cell imaging) and 3D assays (nanofibre plates, collagen spheroid invasion).
- Examined the effects of LiCl and BIO on cell migration and cytoskeletal dynamics.
Main Results:
- All tested paediatric glioma cell lines exhibited migratory potential.
- LiCl and BIO significantly reduced cell migration across all lines.
- Immunofluorescence and live cell imaging revealed drug-induced cytoskeletal rearrangement, loss of polarity, and altered cellular movement.
Conclusions:
- This study is the first to demonstrate the in vitro inhibition of paediatric glioma cell migration using LiCl and BIO.
- These small molecules show promise as potential anti-migratory agents for paediatric brain tumours.
- Further preclinical research into LiCl, BIO, and their derivatives is warranted for therapeutic development.
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