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( A ) In vivo CCS1477 treatment schematic for survival analyses. <t>CD45.2</t> + D/F or T/F AML cells were transplanted into sublethally <t>irradiated</t> <t>CD45.1</t> + WT recipient mice. Treatment was initiated 14 days posttransplant. CCS1477 (20 mg/kg) or vehicle was dosed by mouth (PO) once daily (QD). ( B ) Average ± SEM percent c-Kit + cells in the CD45.2 + peripheral blood (PB) of CCS1477- and vehicle-treated D/F ( n = 7 per treatment) and T/F ( n = 11 per treatment) transplanted AML mice over 2 weeks. Week 0 time point was immediately before the first treatment, followed by weeks 1 and 2 on treatment. Significant differences were evaluated by two-way analysis of variance (ANOVA) Šídák’s multiple comparisons test. Average ± SEM ( C ) spleen weight, and numbers of ( D ) CD45.2 + and ( E ) CD45.2 + LSK (Lin − Sca1 + Kit + ) bone marrow cells of D/F ( n = 8 per treatment) or T/F ( n = 8 per treatment) AML mice after 2 weeks of CCS1477 or vehicle. Significant differences were evaluated by unpaired t test. ( F ) Kaplan-Meier survival analysis of CCS1477-treated and vehicle-treated D/F AML (CCS1477, n = 12; vehicle, n = 14) and T/F AML (CCS1477, n = 9; vehicle, n = 9) transplant recipient mice. Overall survival (OS) of moribund D/F AML mice was 94 and 73 days for CCS1477 and vehicle, respectively. OS of moribund T/F AML mice was 51 and 39 days for CCS1477 and vehicle, respectively. Significance determined by the log-rank (Mantel-Cox) test. ( G ) Average ± SEM spleen weight at mouse moribundity of CCS1477- or vehicle-treated D/F ( n = 3 per treatment) or T/F ( n = 3 per treatment) AML mice. Significant differences were evaluated by unpaired t test. Individual data points represent biological replicates. For all panels, * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001.
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( A ) In vivo CCS1477 treatment schematic for survival analyses. <t>CD45.2</t> + D/F or T/F AML cells were transplanted into sublethally <t>irradiated</t> <t>CD45.1</t> + WT recipient mice. Treatment was initiated 14 days posttransplant. CCS1477 (20 mg/kg) or vehicle was dosed by mouth (PO) once daily (QD). ( B ) Average ± SEM percent c-Kit + cells in the CD45.2 + peripheral blood (PB) of CCS1477- and vehicle-treated D/F ( n = 7 per treatment) and T/F ( n = 11 per treatment) transplanted AML mice over 2 weeks. Week 0 time point was immediately before the first treatment, followed by weeks 1 and 2 on treatment. Significant differences were evaluated by two-way analysis of variance (ANOVA) Šídák’s multiple comparisons test. Average ± SEM ( C ) spleen weight, and numbers of ( D ) CD45.2 + and ( E ) CD45.2 + LSK (Lin − Sca1 + Kit + ) bone marrow cells of D/F ( n = 8 per treatment) or T/F ( n = 8 per treatment) AML mice after 2 weeks of CCS1477 or vehicle. Significant differences were evaluated by unpaired t test. ( F ) Kaplan-Meier survival analysis of CCS1477-treated and vehicle-treated D/F AML (CCS1477, n = 12; vehicle, n = 14) and T/F AML (CCS1477, n = 9; vehicle, n = 9) transplant recipient mice. Overall survival (OS) of moribund D/F AML mice was 94 and 73 days for CCS1477 and vehicle, respectively. OS of moribund T/F AML mice was 51 and 39 days for CCS1477 and vehicle, respectively. Significance determined by the log-rank (Mantel-Cox) test. ( G ) Average ± SEM spleen weight at mouse moribundity of CCS1477- or vehicle-treated D/F ( n = 3 per treatment) or T/F ( n = 3 per treatment) AML mice. Significant differences were evaluated by unpaired t test. Individual data points represent biological replicates. For all panels, * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001.
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( A ) In vivo CCS1477 treatment schematic for survival analyses. CD45.2 + D/F or T/F AML cells were transplanted into sublethally irradiated CD45.1 + WT recipient mice. Treatment was initiated 14 days posttransplant. CCS1477 (20 mg/kg) or vehicle was dosed by mouth (PO) once daily (QD). ( B ) Average ± SEM percent c-Kit + cells in the CD45.2 + peripheral blood (PB) of CCS1477- and vehicle-treated D/F ( n = 7 per treatment) and T/F ( n = 11 per treatment) transplanted AML mice over 2 weeks. Week 0 time point was immediately before the first treatment, followed by weeks 1 and 2 on treatment. Significant differences were evaluated by two-way analysis of variance (ANOVA) Šídák’s multiple comparisons test. Average ± SEM ( C ) spleen weight, and numbers of ( D ) CD45.2 + and ( E ) CD45.2 + LSK (Lin − Sca1 + Kit + ) bone marrow cells of D/F ( n = 8 per treatment) or T/F ( n = 8 per treatment) AML mice after 2 weeks of CCS1477 or vehicle. Significant differences were evaluated by unpaired t test. ( F ) Kaplan-Meier survival analysis of CCS1477-treated and vehicle-treated D/F AML (CCS1477, n = 12; vehicle, n = 14) and T/F AML (CCS1477, n = 9; vehicle, n = 9) transplant recipient mice. Overall survival (OS) of moribund D/F AML mice was 94 and 73 days for CCS1477 and vehicle, respectively. OS of moribund T/F AML mice was 51 and 39 days for CCS1477 and vehicle, respectively. Significance determined by the log-rank (Mantel-Cox) test. ( G ) Average ± SEM spleen weight at mouse moribundity of CCS1477- or vehicle-treated D/F ( n = 3 per treatment) or T/F ( n = 3 per treatment) AML mice. Significant differences were evaluated by unpaired t test. Individual data points represent biological replicates. For all panels, * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001.

Journal: Science Advances

Article Title: P300/CBP inhibition with inobrodib in combination with gilteritinib and venetoclax targets leukemia stem cells in epigenetic mutant AML

doi: 10.1126/sciadv.aec9305

Figure Lengend Snippet: ( A ) In vivo CCS1477 treatment schematic for survival analyses. CD45.2 + D/F or T/F AML cells were transplanted into sublethally irradiated CD45.1 + WT recipient mice. Treatment was initiated 14 days posttransplant. CCS1477 (20 mg/kg) or vehicle was dosed by mouth (PO) once daily (QD). ( B ) Average ± SEM percent c-Kit + cells in the CD45.2 + peripheral blood (PB) of CCS1477- and vehicle-treated D/F ( n = 7 per treatment) and T/F ( n = 11 per treatment) transplanted AML mice over 2 weeks. Week 0 time point was immediately before the first treatment, followed by weeks 1 and 2 on treatment. Significant differences were evaluated by two-way analysis of variance (ANOVA) Šídák’s multiple comparisons test. Average ± SEM ( C ) spleen weight, and numbers of ( D ) CD45.2 + and ( E ) CD45.2 + LSK (Lin − Sca1 + Kit + ) bone marrow cells of D/F ( n = 8 per treatment) or T/F ( n = 8 per treatment) AML mice after 2 weeks of CCS1477 or vehicle. Significant differences were evaluated by unpaired t test. ( F ) Kaplan-Meier survival analysis of CCS1477-treated and vehicle-treated D/F AML (CCS1477, n = 12; vehicle, n = 14) and T/F AML (CCS1477, n = 9; vehicle, n = 9) transplant recipient mice. Overall survival (OS) of moribund D/F AML mice was 94 and 73 days for CCS1477 and vehicle, respectively. OS of moribund T/F AML mice was 51 and 39 days for CCS1477 and vehicle, respectively. Significance determined by the log-rank (Mantel-Cox) test. ( G ) Average ± SEM spleen weight at mouse moribundity of CCS1477- or vehicle-treated D/F ( n = 3 per treatment) or T/F ( n = 3 per treatment) AML mice. Significant differences were evaluated by unpaired t test. Individual data points represent biological replicates. For all panels, * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001.

Article Snippet: For CCS1477 monotherapy in vivo studies, CD45.1 + C57BL6/J mice (Charles River Laboratories), 7 to 8 weeks of age, were irradiated with 500 rads before transplantation with either 10 million CD45.2 + D/F AML cells or 2.5 million CD45.2 + T/F AML cells via retro-orbital injection.

Techniques: In Vivo, Irradiation