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Updated: Aug 8, 2025
A Protocol for Rapid Post-mortem Cell Culture of Diffuse Intrinsic Pontine Glioma DIPG
Published on: March 7, 2017
Cooperativity between H3.3K27M and PDGFRA poses multiple therapeutic vulnerabilities in human iPSC-derived diffuse
Kasey R Skinner1,2,3, Tomoyuki Koga4,3, Shunichiro Miki5,3
1Division of Neuropathology, Department of Pathology, O'Neal Comprehensive Cancer Center and Comprehensive Neuroscience Center, Heersink School of Medicine, University of Alabama at Birmingham, Birmingham, AL 35294, USA.
Abstract:
Diffuse midline glioma (DMG) is a leading cause of brain tumor death in children. In addition to hallmark H3.3K27M mutations, significant subsets also harbor alterations of other genes, such as TP53 and PDGFRA. Despite the prevalence of H3.3K27M, the results of clinical trials in DMG have been mixed, possibly due to the lack of models recapitulating its genetic heterogeneity. To address this gap, we developed human iPSC-derived tumor models harboring TP53R248Q with or without heterozygous H3.3K27M and/or PDGFRAD842V overexpression. The combination of H3.3K27M and PDGFRAD842V resulted in more proliferative tumors when gene-edited neural progenitor (NP) cells were implanted into mouse brains compared to NP with either mutation alone. Transcriptomic comparison of tumors and their NP cells of origin identified conserved JAK/STAT pathway activation across genotypes as characteristic of malignant transformation. Conversely, integrated genome-wide epigenomic and transcriptomic analyses, as well as rational pharmacologic inhibition, revealed targetable vulnerabilities unique to the TP53R248Q; H3.3K27M; PDGFRAD842V tumors and related to their aggressive growth phenotype. These include AREG-mediated cell cycle control, altered metabolism, and vulnerability to combination ONC201/trametinib treatment. Taken together, these data suggest that cooperation between H3.3K27M and PDGFRA influences tumor biology, underscoring the need for better molecular stratification in DMG clinical trials.
Insights
Diffuse midline glioma (DMG) models reveal H3.3K27M and PDGFRA cooperation drives tumor growth. Targeting unique vulnerabilities like AREG offers new therapeutic strategies for pediatric brain tumors.
Area of Science:
- Pediatric neuro-oncology
- Cancer genomics and epigenomics
- Stem cell-derived disease modeling
Background:
- Diffuse midline glioma (DMG) is a fatal pediatric brain tumor with limited treatment options.
- Existing models fail to capture the genetic heterogeneity of DMG, hindering clinical trial success.
- Hallmark H3.3K27M mutations coexist with other genetic alterations like TP53 and PDGFRA in DMG.
Approach:
- Developed patient-derived induced pluripotent stem cell (iPSC) models of DMG.
- Incorporated TP53R248Q, H3.3K27M, and PDGFRAD842V mutations individually and in combination.
- Implanted gene-edited neural progenitor (NP) cells into mouse brains to assess tumor formation and proliferation.
Key Points:
- Combined H3.3K27M and PDGFRAD842V mutations accelerated tumor proliferation in iPSC-derived models.
- JAK/STAT pathway activation was conserved across genotypes during malignant transformation.
- Integrated multi-omics analyses identified unique vulnerabilities in triple-mutant DMG, including AREG-mediated cell cycle control and metabolic alterations.
Conclusions:
- Cooperation between H3.3K27M and PDGFRA drives DMG progression.
- Targeting specific vulnerabilities, such as AREG, and combination therapies (ONC201/trametinib) show promise.
- Improved molecular stratification is crucial for advancing DMG clinical trials.
