Characterization of the Differentiation Patterns of Histone H3 K27M Neural Stem Cells
Engineering
BME195
This project seeks to characterize the nature of a hypothesized differentiation stall induced by the Histone H3 K27M mutation in neural stem cells. This mutation occurs in 30% of pediatric high-grade gliomas and 80% of Diffuse Intrinsic Pontine Gliomas, a more aggressive form of pediatric high-grade gliomas. Investigation of the cell type specific timing of the differentiation stall in the neural development pathway was conducted with immunofluorescence staining of cerebral organoids for the following protein markers: SOX2 (undifferentiated pluripotent stem cells, neural epithelium and radial glia neural stem cells), SATB2 (upper layer neurons), VIM (radial glia neural stem cells), TBR2 (intermediate progenitor cells), PAX6 (radial glia cells), CTIP2 (deep layer neurons), and RELN (layer 1 cajal retzius neurons). To supplement this visual data, single-cell RNA sequencing analysis was used to further examine the cell types present in cerebral organoids harboring the H3 K27M Mutation. Analysis of both sets of data supports the hypothesis that the differentiation stall occurs between neuroepithelium and radial glia cells but further optimization of the single-cell RNA sequencing pipeline is ongoing to provide a more accurate location of the differentiation stall.
