HGG-12_Kasey Skinner


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Kasey Skinner1,2, Tomoyuki Koga3, Shunichiro Miki4,5, Robert F. Gruener6, R. Stephanie Huang7, Frank Furnari4,5, C. Ryan Miller2;

1Neuroscience Curriculum, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA. 2Department of Pathology, University of Alabama Birmingham, Birmingham, AL, USA. 3Department of Neurosurgery, University of Minnesota Medical School, Minneapolis, MN, USA. 4Ludwig Institute for Cancer Research, La Jolla, CA, USA. 5Department of Pathology, University of California at San Diego, La Jolla, CA, USA. 6Ben May Department for Cancer Research, University of Chicago, Chicago, IL, USA. 7Department of Experimental and Clinical Pharmacology, University of Minnesota, Minneapolis, MN, USA

Diffuse intrinsic pontine glioma (DIPG) is a subset of high-grade glioma that occurs predominantly in children and has no cure. Up to 80% of DIPG harbor a heterozygous point mutation that results in a lysine 27 to methionine substitution in histone variant H3.3 (H3.3K27M). Existing DIPG models have provided insight into the role of H3.3K27M but have limitations: genetically engineered murine models often rely on overexpression of the mutant histone to form tumors; patient-derived xenografts (PDX) are more genetically faithful but preclude examination of the effect of individual mutations on pathogenesis. To address these shortcomings and better recapitulate the genetics of human tumors, we designed a novel DIPG model based on human induced pluripotent stem cells (iPSC) edited via CRISPR to express heterozygous H3.3K27M. Edited iPSC were chemically differentiated into neural progenitor cells, which upon implantation into the brainstems of immunodeficient mice formed diffusely invasive tumors that were histologically consistent with high-grade glioma. Further, neurospheres cultured from primary tumors formed secondary tumors upon reimplantation with more diffuse invasion, suggesting in vivo evolution. To validate this model’s relevance to DIPG transcriptionally, we performed RNA-sequencing on a cohort of primary and secondary tumor neurospheres (termed primary and secondary iDIPG) and compared them to published RNA-seq data from pediatric PDX and patient tumor samples. Hierarchical clustering and principal component analysis on differentially expressed genes (P<0.05) showed that H3.3K27M iDIPG cluster with H3.3K27M PDX and patient tumors. Further, ssGSEA showed that H3.3K27M iDIPG are enriched for astrocytic and mesenchymal signature genes, a defining feature of H3.3K27M DIPG. Finally, we found that primary H3.3K27M iDIPG neurospheres are sensitive to panobinostat, an HDAC inhibitor shown to be effective against H3.3K27M DIPG cells in vitro. Overall, these data suggest that H3.3K27M iDIPG are a promising tool for investigating DIPG biology and new therapeutic strategies.

Duration: 04:59

Posted: Sunday, June 6, 2021

Video tags: 2021 SNO Pediatric Meeting