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Updated: Jun 21, 2025
Laboratory-Engineered Glioblastoma Organoid Culture and Drug Screening
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Parsing the effect of co-culture with brain organoids on Diffuse Intrinsic Pontine Glioma (DIPG) using quantitative
Victoria G Prior1, Simon Maksour2, Sara Miellet2
1Children's Cancer Research Unit, Kids Research, The Children's Hospital at Westmead, Westmead, Australia; The University of Sydney, Discipline of Child and Adolescent Health, The Children's Hospital at Westmead, Westmead, Australia.
Abstract:
Diffuse Intrinsic Pontine Gliomas (DIPGs) are deadly brain cancers in children for which there is no effective treatment. This can partly be attributed to preclinical models that lack essential elements of the in vivo tissue environment, resulting in treatments that appear promising preclinically, but fail to result in effective cures. Recently developed co-culture models combining stem cell-derived brain organoids with brain cancer cells provide tissue dimensionality and a human-relevant tissue-like microenvironment. As these models are technically challenging, we aimed to establish whether interaction with the organoid influences DIPG biology and thus warrants their use. To address this question DIPG24 cells were cultured with pluripotent stem cell-derived cortical organoids. We created "mosaic" co-cultures enriched for tumour cell-neuronal cell interactions versus "assembloid" co-cultures enriched for tumour cell-tumour cell interactions. Sequential window acquisition of all theoretical mass spectra (SWATH-MS) was used to analyse the proteomes of DIPG fractions isolated by flow-assisted cell sorting. Control proteomes from DIPG spheroids were compared with DIPG cells isolated from mosaic and assembloid co-cultures. This suggested changes in cell interaction with the external environment reflected by decreased gene ontology terms associated with adhesion and extracellular matrix, and increased DNA synthesis and replication, in DIPG24 cells under either co-culture condition. By contrast, the mosaic co-culture was associated with neuron-specific brahma-associated factor (nBAF) complex signalling, a process associated with neuronal maturation. We propose that co-culture with brain organoids is a valuable tool to parse the contribution of the brain microenvironment to DIPG tumour biology.
Insights
Co-culturing Diffuse Intrinsic Pontine Glioma (DIPG) cells with brain organoids reveals microenvironmental influences on tumor biology. This approach, using novel co-culture models, provides insights into DIPG progression and potential therapeutic strategies.
Area of Science:
- Neuro-oncology
- Developmental neurobiology
- Cancer biology
Background:
- Diffuse Intrinsic Pontine Gliomas (DIPGs) are aggressive pediatric brain tumors with no effective treatments.
- Current preclinical models inadequately replicate the in vivo tumor microenvironment, leading to translational failures.
- Stem cell-derived brain organoids offer a more human-relevant microenvironment for studying complex diseases.
Purpose of the Study:
- To investigate the impact of brain organoid co-culture on Diffuse Intrinsic Pontine Glioma (DIPG) cell biology.
- To determine if co-culture models warrant further development for DIPG research.
- To analyze proteomic changes in DIPG cells when interacting with neuronal organoids.
Main Methods:
- DIPG24 cells were co-cultured with pluripotent stem cell-derived cortical organoids, creating "mosaic" (tumor-neuron) and "assembloid" (tumor-tumor) models.
- Proteomic analysis of isolated DIPG cells was performed using Sequential Window Acquisition of All Theoretical Mass Spectra (SWATH-MS).
- Comparative proteomic analysis was conducted between DIPG spheroids and DIPG cells from co-culture models.
Main Results:
- DIPG24 cells in co-culture exhibited altered cellular processes, including decreased adhesion and extracellular matrix interactions, and increased DNA synthesis and replication.
- Mosaic co-cultures specifically showed enrichment of neuron-specific brahma-associated factor (nBAF) complex signaling, linked to neuronal maturation.
- Proteomic shifts suggest the brain microenvironment significantly influences DIPG cell behavior.
Conclusions:
- Co-culture with brain organoids provides a valuable platform for dissecting the influence of the brain microenvironment on DIPG tumor biology.
- These advanced models can reveal mechanisms of DIPG progression not evident in simpler preclinical systems.
- Further development of organoid co-culture models holds promise for advancing DIPG research and therapeutic development.
