Generation of diffuse intrinsic pontine glioma mouse models by brainstem targeted in utero electroporation

Abstract

Background Diffuse intrinsic pontine gliomas (DIPGs) are highly lethal childhood brain tumors. Their unique genetic makeup, pathological heterogeneity, and brainstem location all present challenges to treatment. Developing mouse models that accurately reflect each of these distinct features will be critical to advance our understanding of DIPG development, progression, and therapeutic resistance. The aim of this study was to generate new mouse models of DIPG, and characterize the role of specific oncogenic combinations in DIPG pathogenesis. Methods We used in utero electroporation (IUE) to transfect neural stem cells in the developing brainstem with PiggyBac DNA transposon plasmids. Combinations of PDGFB, PdgfraD842V, or PdgfraWT, combined with dominant negative Trp53 (DNp53) and H3.3K27M expression induced fully penetrant brainstem gliomas. Results IUE enabled the targeted transfection of brainstem neural stem cells. PDGFB + DNp53 + H3.3K27M induced the rapid development of grade-IV gliomas. PdgfraD842V + DNp53 + H3.3K27M produced slower forming grade-III gliomas. PdgfraWT + DNp53 + H3.3K27M produced high and low-grade gliomas with extended latencies. PDGFB, PdgfraD842V, and PdgfraWT DIPG models display unique histopathological and molecular features found in human DIPGs. Conclusion Brainstem targeted in utero electroporation provides a rapid and flexible system to generate diverse DIPG mouse models. Using IUE to investigate mutation and pathohistological heterogeneity of DIPG will provide a valuable tool for future genetic and preclinical studies.

Everolimus improves the efficacy of dasatinib in PDGFRα-driven glioma

“…In order to study the utility of dasatinib in PDGFRα-driven HGG, we adapted an intrauterine electroporation (IUE) mouse model (ref. 15 and Figure 1A). We induced glioma in mice by injecting plasmids encoding: (a) PBase; (b) PB-CAG-DNp53-Ires-luciferase (TP53); (c) PB-CAG-PdgfraD824V-Ires-EGFP (PDGFRA D842V); and (d) PB-CAG-H3.3 K27M-Ires-EGFP (H3K27M) into the lateral ventricles (forebrain) of E13.5 embryos in CD1 mice.…”

“…Murine model of HGG using IUE IUE was performed using sterile technique on isoflurane/oxygenanesthetized pregnant CD1 female mice at E13.5 (cortex), according to established methodology (36). All tumors for this study were generated by injecting plasmids into either the lateral ventricle (forebrain) or the fourth ventricle (hindbrain).…”

Histone-Mutant Glioma: Molecular Mechanisms, Preclinical Models, and Implications for Therapy

“…Other IUE-based models have directly compared the effects of exogenous PDGF ligand versus receptor. The combination of piggyBac transposon-mediated embryonic IUE of H3.3K27M, p53 loss, and either PDGF-B, PDGFRA WT , or constitutive mutant PDGFRA D842V all resulted in fully penetrant glioma formation, but with distinct differences in histological characterization [ 32 ]. Exogenous PDGF ligand led to cell-extrinsic perivascular changes that contributed to the shortest latency, while WT PDGFRA led to less aggressive tumors with longer latency as compared with PDGFRA D842V .…”

Clinical efficacy and predictive biomarkers of ONC201 in H3 K27M-mutant diffuse midline glioma

“…1a). To evaluate the in vivo e cacy of ONC201, we adopted a midline in utero electroporation (IUE) mouse model of H3 K27M-glioma 10,11 , in which tumors harboring dominant negative TP53, PDGFRA D842V, and H3F3A (H3.3) K27M mutations ("PPK") are generated ( Fig. 1b-c).…”