Fig. 6.

ONC201 and ONC206 single and combination treatments induce transcriptional reprogramming and cell lineage shifts. (A) DMG cells were treated with ONC201/6 as single or in combination (48 hrs), followed by bulk RNA sequencing. Differential gene expression (volcano plot) of cells treated with monotherapy (ONC201, top left; ONC206, top right), and combination therapy (bottom) indicated differential gene expression induced by each drug. (B) Combination treatment resulted in significant differential gene expression (relative to DMSO control). Functional enrichment analyses (Gene Ontology, KEGG) identfied molecular pathways uniquely activated (red) and inhibited (blue) in each condition. (C) Left panel: Single-cell RNA sequencing of DMG cells treated with ONC201 (96 hrs) showed a decrease in cycling and oligodendrocyte-like progenitor cells (OPC), and an increase in differentiated astrocytic-like (AC) cells. PCA plot (left) and bar graph (right) show the proportion of cells (AC, OPC, oligodendrocyte-like, cycling) in control (DMSO) and in treated conditions. Right panel: Visual representation of cell lineage shift from OPC-like to AC-like cells in response to imipridone treatment. Arrows from OPC towards AC or oligodendrocyte lineages indicate developmental hierarchy; arrow encircling cycling cells indicates self-renewal and replenishment of OPC pool. Area taken up by a given cell type is respective to its observed proportion in the left panel.