Figure 3.
HDAC inhibitor and H3K9 methyltransferase inhibition reversed the epigenetic abnormality and inhibited maintenance of Dnmt3a−/−;Idh2R140Qleukemia. (A) Western blot of histone modification in c-kit+ bone marrow progenitor cells of mice from all genotypes. (B) Global histone modification shows a decrease of H3K9me3 and an increase of H3K9/27ac after HDAC inhibitor treatment in bone marrow cells from 3aKO-140 mice in vitro. (C) HDAC inhibitor treatment (vorinostat) reduced the proliferation of double-mutant cells in a colony-forming assay at day 7. Mean ± standard deviation (SD); the results of 3 independent cultures are presented. *P < .05; **P < .01, by Student t test. (D-G) HDAC inhibitor treatment reduced leukocytosis (D-E) and reversed anemia (F) and thrombocytopenia (G) in the recipients of Dnmt3a−/−;Idh2R140Q leukemic cells. Values are means ± SD (n = 7 and 6 in the vehicle- and inhibitor-treated groups, respectively). **P < .01, ***P < .001, by Student t test analysis. (H) Representative blood smears for vehicle- and panobinostat-treated double-mutant recipient mice. (I) The flow plot of mouse bone marrow treated with panobinostat and DMSO. The mouse bone marrow progenitor (c-kit+, Mac1&Gr1−) is gated. (J) Kaplan-Meier survival curves for mice treated with DMSO and panobinostat (10 mg/kg). **P < .01, by long-rank test. Inset: a reduction of splenomegaly after treatment with panobinostat (10 mg/kg) at 4 weeks after bone marrow transplantation. (K) Response of 3aKO-140 HSPCs to the Ehmt2 inhibitor BIX01294 in in vitro colony-forming assay of 1000 c-kit+ cells (left). Immunoblot for H3K9me2 of HSPCs with DMSO or BIX01294 treatment (right). (L) Response of 3aKO-140 HSPCs to the Ezh2 inhibitors EPZ-6438 and GSK343 in in vitro colony-forming assay of 1000 c-kit+ cells (left and middle). Immunoblot for H3K27me3 of HSPCs with EPZ-6438 or GSK343 treatment (right). (M) The colony forming units of Dnmt3aKO c-kit+ bone marrow cells under G9a inhibitor Bix01294, HDAC inhibitor vorinostat and EZH2 inhibitor. Mean ± SD (n = 3 per group). (N), The CFU of Idh2R140Q c-kit+ bone marrow cells under G9a inhibitor Bix01294, HDAC inhibitor vorinostat and EZH2 inhibitor. Mean ± SD (n = 3 per group).

HDAC inhibitor and H3K9 methyltransferase inhibition reversed the epigenetic abnormality and inhibited maintenance of Dnmt3a−/−;Idh2R140Qleukemia. (A) Western blot of histone modification in c-kit+ bone marrow progenitor cells of mice from all genotypes. (B) Global histone modification shows a decrease of H3K9me3 and an increase of H3K9/27ac after HDAC inhibitor treatment in bone marrow cells from 3aKO-140 mice in vitro. (C) HDAC inhibitor treatment (vorinostat) reduced the proliferation of double-mutant cells in a colony-forming assay at day 7. Mean ± standard deviation (SD); the results of 3 independent cultures are presented. *P < .05; **P < .01, by Student t test. (D-G) HDAC inhibitor treatment reduced leukocytosis (D-E) and reversed anemia (F) and thrombocytopenia (G) in the recipients of Dnmt3a−/−;Idh2R140Q leukemic cells. Values are means ± SD (n = 7 and 6 in the vehicle- and inhibitor-treated groups, respectively). **P < .01, ***P < .001, by Student t test analysis. (H) Representative blood smears for vehicle- and panobinostat-treated double-mutant recipient mice. (I) The flow plot of mouse bone marrow treated with panobinostat and DMSO. The mouse bone marrow progenitor (c-kit+, Mac1&Gr1−) is gated. (J) Kaplan-Meier survival curves for mice treated with DMSO and panobinostat (10 mg/kg). **P < .01, by long-rank test. Inset: a reduction of splenomegaly after treatment with panobinostat (10 mg/kg) at 4 weeks after bone marrow transplantation. (K) Response of 3aKO-140 HSPCs to the Ehmt2 inhibitor BIX01294 in in vitro colony-forming assay of 1000 c-kit+ cells (left). Immunoblot for H3K9me2 of HSPCs with DMSO or BIX01294 treatment (right). (L) Response of 3aKO-140 HSPCs to the Ezh2 inhibitors EPZ-6438 and GSK343 in in vitro colony-forming assay of 1000 c-kit+ cells (left and middle). Immunoblot for H3K27me3 of HSPCs with EPZ-6438 or GSK343 treatment (right). (M) The colony forming units of Dnmt3aKO c-kit+ bone marrow cells under G9a inhibitor Bix01294, HDAC inhibitor vorinostat and EZH2 inhibitor. Mean ± SD (n = 3 per group). (N), The CFU of Idh2R140Q c-kit+ bone marrow cells under G9a inhibitor Bix01294, HDAC inhibitor vorinostat and EZH2 inhibitor. Mean ± SD (n = 3 per group).

Close Modal
This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal