Journal: iScience

Article Title: Regulatory role of mTORC1 signaling in osteoblasts in acute myeloid leukemia progression and steady-state hematopoiesis

doi: 10.1016/j.isci.2025.114533

Figure Lengend Snippet: mTORC1 activation in osteoblasts accelerates AML progression (A) Schematic diagram of generation of tissue-specific Tsc1 knockout mice. (B) Schematic diagram of generation of a MLL-AF9 murine AML model. (C and D) Representative flow cytometry plots and percentages of GFP + AML cells in the (C) peripheral blood ( n = 7–9) and (D) bone marrow ( n = 11) of Tsc1 fl/fl and Col1a1-Cre;Tsc1 fl/fl mice. (E–G) Representative flow cytometry plots and percentages of (E) GFP + c-Kit – AML cells and GFP + c-Kit + AML cells ( n = 6–7), (F) BrdU + GFP + c-Kit + AML cells ( n = 4–5), and (G) Annexin V + GFP + c-Kit + AML cells ( n = 4–5) in the bone marrow of Tsc1 fl/fl and Col1a1-Cre;Tsc1 fl/fl mice. (H) Percentages of GFP + AML cells in the peripheral blood and bone marrow of Tsc1 fl/fl and Col1a1-Cre;Tsc1 fl/fl mice 8 days after the second transplantation ( n = 3). (I and J) Survival probabilities of (I) Tsc1 fl/fl and Col1a1-Cre;Tsc1 fl/fl AML mice ( n = 6–7) and (J) WT AML mice transplanted with c-Kit + AML cells from Raptor fl/fl and Col1a1-Cre;Raptor fl/fl mice ( n = 5–7). All mice used in this study were male. n.s., not significant. ∗ p < 0.05. Error bars show the standard deviation.

Article Snippet: Tsc1 fl/fl mice , Jackson Laboratory , #005680.

Techniques: Activation Assay, Knock-Out, Flow Cytometry, Transplantation Assay, Standard Deviation

Journal: iScience

Article Title: Regulatory role of mTORC1 signaling in osteoblasts in acute myeloid leukemia progression and steady-state hematopoiesis

doi: 10.1016/j.isci.2025.114533

Figure Lengend Snippet: Activation of mTORC1 in osteoblasts impairs normal hematopoiesis (A) Schematic diagram of flow cytometric analysis of Tsc1 fl/fl and Col1a1-Cre;Tsc1 fl/fl mice. (B) The number of BM-MNCs in hind legs of Tsc1 fl/fl and Col1a1-Cre;Tsc1 fl/fl mice ( n = 9–10). (C–F) Representative flow cytometry plots and percentages of (C) LSK cells and LSK subpopulations ( n = 17–18), (D) LK cells and LK subpopulations ( n = 14–15), (E) CD11b + Gr-1 + myeloid cells ( n = 15–16), and (F) B220 + IgM – immature and B220 + IgM + mature B cells ( n = 16–18) in the bone marrow of Tsc1 fl/fl and Col1a1-Cre;Tsc1 fl/fl mice. (G) Representative flow cytometry plots and fold changes in the percentage of LSK cells ( n = 7–18), CD11b + Gr-1 + myeloid cells ( n = 5–18), and B220 + IgM – immature and B220 + IgM + mature B cells ( n = 5–16) in the bone marrow of Tsc1 fl/fl , Col1a1-Cre;Tsc1 fl/fl , and Col1a1-Cre;Tsc1 fl/fl ;Raptor fl/+ mice. All mice used in this study were male. n.s., not significant. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, # p < 0.05, ## p < 0.01, and ### p < 0.001. Error bars show the standard deviation.

Article Snippet: Tsc1 fl/fl mice , Jackson Laboratory , #005680.

Techniques: Activation Assay, Flow Cytometry, Standard Deviation

Journal: iScience

Article Title: Regulatory role of mTORC1 signaling in osteoblasts in acute myeloid leukemia progression and steady-state hematopoiesis

doi: 10.1016/j.isci.2025.114533

Figure Lengend Snippet: IL-6 in osteoblasts plays a crucial role for AML progression (A) Schematic diagram of three datasets used to screen for factors involved in the cell-cell interaction between mTORC1-activated osteoblasts and AML cells. (B and C) Volcano plots showing DEGs (B) in osteoblasts co-cultured with AML cells compared to monocultured osteoblasts and (C) in mTORC1 high osteoblasts compared to mTORC1 low osteoblasts. (D) Venn diagram highlighting overlapping genes identified across the three datasets. (E) Il6 mRNA expression in calvarial osteoblasts from Col1a1-Cre;Tsc1 fl/fl mice ( n = 4–5). (F) GSEA results for the HALLMARK_IL6_JAK_STAT3_SIGNALING gene set in AML cells co-cultured with osteoblasts. (G) MFI of phosphorylated STAT3 in AML cells in the murine AML model and in lineage – cells from WT mice ( n = 4–5). (H) Schematic diagram of generation of the IL-6R-knockdown MLL-AF9 murine AML model. (I) Survival probabilities of Tsc1 fl/fl and Col1a1-Cre;Tsc1 fl/fl mice transplanted with shCtrl- or sh Il6r -transduced AML cells ( n = 10–20). (J) Schematic model of the findings of this study. Osteoblastic mTORC1 signaling enhances IL-6 production, which activates JAK/STAT3 signaling to promote cell proliferation and inhibit apoptosis in undifferentiated AML cells, driving AML progression. All mice used in this study were male. n.s., not significant. ∗ p < 0.05, ∗∗∗ p < 0.001, and # p < 0.05. Error bars show the standard deviation.

Article Snippet: Tsc1 fl/fl mice , Jackson Laboratory , #005680.

Techniques: Cell Culture, Expressing, Knockdown, Standard Deviation