Fig. 2

Insights in biological and molecular functions of miR-17–92 cluster in T-ALL. A Schematic diagram of the approach used for PDX T-ALLs transduced with lentiviral constructs encoding miR-17–92 cluster or empty as a control (Empty V.) and cultured for 3 days on MS5-DL1 feeders. Transduced (GFP +) cells were then sorted with FACS and transplanted into immunodeficient NSG recipient mice, which were subsequently treated with DAPT γ-secretase inhibitor (1 mg/mouse) or dimethyl sulfoxide (DMSO), both delivered by intraperitoneal injection at days 4 and 8 post-transplant. B Flow cytometry analysis of GFP + CD45 + alive cells in peripheral blood, bone marrow or spleen from transplanted recipient mice treated as describe in (A). Y-axes, percentage of GFP + cells. X-axes, experimental conditions. Significance analysis was performed by Mann–Whitney U test. C Graphical representation of canonical (Mod-A) and of miR-17–92 modulated NOTCH1-signalling pathway (Mod-B). D Hierarchical clustering analysis of Mod-A and Mod-B gene expression profile in the various experimental conditions (i.e., ± GSI; miR-17–92 OE or empty vector). Main clusters of genes are also indicated as GSI-UP/GSI-DOWN (Mod-A) or miR-UP/miR-DOWN (Mod-B). Colours are as per the legend. On top, Venn diagram showing Mod-A/B number of genes and relative overlapping. E Percentage distribution of ‘ETP status’ and ‘Maturation stage’ of T-ALLs in the Liu et al. cohort (n = 261) stratified according to ssGSEA using Mod-A (n = 123) and Mod-B (n = 138) gene sets. ETP, Early T-cell Precursor. P-values were computed by chi-square test. F Box-plots show the levels of MRD (at 29 days) and BMA of blasts (at 8 days) in T-ALLs the Liu et al. cohort (n = 261) stratified as in (D). MRD, Minimal Residual Disease. BMA, Bone Marrow Aspirates. G t-SNE plot of scRNAseq data on cell subsets of PDX from T-ALL patients. Colours are as per the legend. H Hierarchical clustering analysis of Enrichment Scores from GSEA using the Mod-A (GSI-UP/DOWN) and Mod-B (miR-UP/miR-DOWN) gene sets in the T-ALL cell subsets profiled by scRNAseq as in (G). I Distributions of enrichment of biological functions which were identified by GSEA using scRNA-expression profiles, in clusters of T-ALL cell subsets as in (H). The higher the size of bubbles the more significant is the enrichment of a particular biofunction. Colors of bubbles indicate the magnitude of normalized enrichment scores (NES) and are as per the legend