Page 110 - Haematologica June

L. Trentin et al.
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Figure 3. Higher leukemia-initiating cell potential of cells in early G1-S transition. (A) Cell cycle “live” staining: simultaneous DNA (Hoechst 33342) and RNA (pyronin Y) labeling of BCP-ALL cells and cell cycle annotated cellular subfractions G1a, G1blow, G1bhigh and G2/M analyzed by FACS. Sorted fractions were transplanted into NOD/SCID mice (105 cells/mouse) and leukemia engraftment was analyzed as weeks from transplantation until appearance of ≥1% huCD19+ ALL cells in peripheral blood of the recipients. (B) Increased engraftment activity upon transplantation of G1blow (red) compared to G2/M (blue) cells; (i, ii) short time to leukemia (TTLshort) ALL, two independent experiments, n=3 and n=4 mice/group; (iii) long time to leukemia (TTLlong) ALL, n=3 mice/group. (C) Maintained distinct engraftment in second- ary recipients transplanted with unsorted bulk primograft ALL derived from primary recipients transplanted with G1blow or G2/M sorted fractions, n=3 and n=2 mice/group, respectively. Log-rank test; P=statistical significance.
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Figure 4. G1blow acute lymphoblastic leukemia cells are characterized by a transcriptional stem cell program. (A) Unsupervised cluster analysis of 865 genes (1122 probe sets) differentially regulated (green: down-regulated; red: up-regulated) between G1blow and G2/M sorted cellular subfractions False discovery rate (FDR) q- val<0.05 (B) Positive (left) enrichment of gene sets attributed to stemness or negative enrichment of sets annotated to mature or short-term stem cells (right) with the G1blow-profile (gene set enrichment analysis, NOM P-value ≤0.05 and FDR q-value ≤0.05).
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haematologica | 2018; 103(6)