Figure 7.
Figure 7. Depletion of CBFA2T3 diminishes LSC gene signatures and in vivo expansion of Kasumi-1 cells. (A) Expression changes of CBFA2T3 cis-actome and cis-repressome genes in CBFA2T3-depleted vs control cells. The boxplot at the right shows all significantly changed CBFA2T3 cis-actome and cis-repressome genes upon depletion of CBFA2T3 in Kasumi-1 cells (P ≤ .05). (B) Heatmap showing increased (green) or decreased (red) expression of genes within the HSC and LSC gene sets indicated at the right in duplicated control and CBFA2T3-knockdown samples. “LSC Up” denotes GAL_LEUKEMIC_STEM_CELL_UP; “LSC Down” denotes GAL_LEUKEMIC_STEM_CELL_DN; “HSC Up” denotes JAATINEN_HEMATOPOIETIC_STEM_CELL_UP; and “HSC Down” denotes JAATINEN_HEMATOPOIETIC_STEM_CELL_DN. CBFA2T3 cis-actome and cis-repressome genes, as well as genes in multiple affected pathways, are marked on the left. (C) Effects of CBFA2T3 knockdown on cell growth of the indicated AML cells. Cells were counted daily starting at 4 days after lentiviral transduction. (D) Flow cytometric, cell cycle quantification of control and CBFA2T3-knockdown t(8;21)-AML cells stained with propidium iodide. Error bars represent the standard error of 3 independent biological replicates. (E) Representative bioluminescence images (left) and average luminescence intensity (right) of control and CBFA2T3-depleted Kasumi-1 cells and SKNO-1 cells engrafted into NSGS (Kasumi-1) or NSG (SKNO-1) mice over the indicated period. 1 × 106 SKNO-1 cells were transplanted into NSG mice 5 hours after radiograph radiation (2.25 Gy); 2.5 × 106 Kasumi-1 cells were transplanted into NSGS mice 5 hours after radiograph radiation (2.5 Gy). (F) Proposed model of the interplay between CBFA2T3, GCN5, and RUNX1-RUNX1T1 in regulating LSC cell fate and AML relapse. The dashed line on the left denotes a possible role for CBFA2T3 in facilitating LSC transformation. The dashed lines on the right denote the abilities of CBFA2T3 to maintain LSCs and inhibit growth arrest, thus facilitating the expansion of LSCs that ultimately lead to relapse. (G) A 2-step classifier predicting AML patient prognosis. Details are given in “Discussion.” *P < .05; **P < .01; ***P < .001.

Depletion of CBFA2T3 diminishes LSC gene signatures and in vivo expansion of Kasumi-1 cells. (A) Expression changes of CBFA2T3 cis-actome and cis-repressome genes in CBFA2T3-depleted vs control cells. The boxplot at the right shows all significantly changed CBFA2T3 cis-actome and cis-repressome genes upon depletion of CBFA2T3 in Kasumi-1 cells (P ≤ .05). (B) Heatmap showing increased (green) or decreased (red) expression of genes within the HSC and LSC gene sets indicated at the right in duplicated control and CBFA2T3-knockdown samples. “LSC Up” denotes GAL_LEUKEMIC_STEM_CELL_UP; “LSC Down” denotes GAL_LEUKEMIC_STEM_CELL_DN; “HSC Up” denotes JAATINEN_HEMATOPOIETIC_STEM_CELL_UP; and “HSC Down” denotes JAATINEN_HEMATOPOIETIC_STEM_CELL_DN. CBFA2T3 cis-actome and cis-repressome genes, as well as genes in multiple affected pathways, are marked on the left. (C) Effects of CBFA2T3 knockdown on cell growth of the indicated AML cells. Cells were counted daily starting at 4 days after lentiviral transduction. (D) Flow cytometric, cell cycle quantification of control and CBFA2T3-knockdown t(8;21)-AML cells stained with propidium iodide. Error bars represent the standard error of 3 independent biological replicates. (E) Representative bioluminescence images (left) and average luminescence intensity (right) of control and CBFA2T3-depleted Kasumi-1 cells and SKNO-1 cells engrafted into NSGS (Kasumi-1) or NSG (SKNO-1) mice over the indicated period. 1 × 106 SKNO-1 cells were transplanted into NSG mice 5 hours after radiograph radiation (2.25 Gy); 2.5 × 106 Kasumi-1 cells were transplanted into NSGS mice 5 hours after radiograph radiation (2.5 Gy). (F) Proposed model of the interplay between CBFA2T3, GCN5, and RUNX1-RUNX1T1 in regulating LSC cell fate and AML relapse. The dashed line on the left denotes a possible role for CBFA2T3 in facilitating LSC transformation. The dashed lines on the right denote the abilities of CBFA2T3 to maintain LSCs and inhibit growth arrest, thus facilitating the expansion of LSCs that ultimately lead to relapse. (G) A 2-step classifier predicting AML patient prognosis. Details are given in “Discussion.” *P < .05; **P < .01; ***P < .001.

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