American Society of Hematology

Figure 6.

$Efficacy of positive hits from stepwise drug discovery approach in vivo. (A) Representative scheme of our stepwise drug discovery approach coupled with in vivo preclinical trials. Tumor cells from T-ALL PDXs were first screened with a library of 433 compounds in 384-well plate format. Positive hits were then validated in 96-well plate format with and without E4-ECs. The number of candidates was narrowed down (n = 29) after implementation of multiple experimental conditions and validations. Representative drugs not rescued by E4-ECs were then tested in vivo and proved to be effective in T-ALL PDX mice. (B) Kaplan-Meier plot depicting overall survival of all the mice (n = 111) treated with vehicle vs different therapeutic regimens across 11 different PDX models. P value was calculated using a log-rank test. (C) Bar plot showing the fold change in the percentage of T-ALL cells in the blood of RO2 and RO6 PDX mice treated with ruxolitinib, bortezomib, tofacitinib, and combinations compared with controls. P values were calculated by t test. ∗P < .05; ∗∗P < .01. (D) Bar plot showing the fold change in the spleen volume (measured by MRI) of RO2 and RO6 mice treated with ruxolitinib, bortezomib, tofacitinib, and combinations compared with controls. P values were calculated by t test. ∗P < .05; ∗∗P < .01. (E) Kaplan-Meier plot depicting overall survival of RO6 mice treated with vehicle, tofacitinib, ruxolitinib, or combination. P value was calculated using a log-rank test. (F) Line-dot plot showing changes in spleen volume (measured by MRI) of 3053 PDX mice treated with irinotecan (20 mg/kg per day) compared with controls in a timeframe of 40 days. P value was calculated by t test. ∗∗∗∗P < .0001. (G) Line-dot plot showing changes in percentage of T-ALL cells in blood of 3053 and 3119 PDX mice treated with irinotecan compared with controls in a timeframe of 80 days. Three of the 12 irinotecan-treated mice relapsed and died of leukemia. P value was calculated by t test. ∗∗∗∗P < .0001. (H) Kaplan-Meier plot depicting overall survival of 3053 and 3119 PDX mice treated with vehicle or irinotecan. Nine of the 12 irinotecan-treated mice died without leukemia. P value was calculated using a log-rank test. (I) Kaplan-Meier plot depicting overall survival of 8 different T-ALL PDX models (6467, 14741, 7072, 7155, 13002, 13356, 10512, and 5384) treated with vehicle or HDAC inhibitor panobinostat (n = 2 mice per arm, n = 32 mice total) (5 mg/kg per day). Mice belonging to 3 of the 8 models (5384, 7155, and 6467) died without leukemia. P value was calculated using a log-rank test. (J) Bar plot depicting the circulating T-ALL cells (difference between day 30 and day 0; day 0: first T-ALL detection in the blood) in 8 PDX models treated with vehicle or panobinostat. A single partially refractory model was identified (1 of 8, 7072 PDX T-ALL). P value was calculated by t test. ∗∗P < .001. (K) Bar plots depicting the percentage of circulating T-ALL cells in 4 PDX models (10512, 6467, 13002, and 13356) treated with vehicle, daunorubicin (1.2 mg/kg for 3 days/week), linsitinib (50 mg/kg per day), or combination (n = 2 mice per arm, n = 32 mice total). Mice treated with daunorubicin plus linsitinib displayed delayed leukemia progression compared with the other arms. P value was calculated by t test. ∗P < .05. (L) Kaplan-Meier plot depicting overall survival of 10512, 6467, 13002, and 13356 T-ALL PDX models treated with vehicle, daunorubicin, linsitinib, or combination. Mice treated with daunorubicin plus linsitinib survived until 3 months from enrollment. P value was calculated using a log-rank test.$

Efficacy of positive hits from stepwise drug discovery approach in vivo. (A) Representative scheme of our stepwise drug discovery approach coupled with in vivo preclinical trials. Tumor cells from T-ALL PDXs were first screened with a library of 433 compounds in 384-well plate format. Positive hits were then validated in 96-well plate format with and without E4-ECs. The number of candidates was narrowed down (n = 29) after implementation of multiple experimental conditions and validations. Representative drugs not rescued by E4-ECs were then tested in vivo and proved to be effective in T-ALL PDX mice. (B) Kaplan-Meier plot depicting overall survival of all the mice (n = 111) treated with vehicle vs different therapeutic regimens across 11 different PDX models. P value was calculated using a log-rank test. (C) Bar plot showing the fold change in the percentage of T-ALL cells in the blood of RO2 and RO6 PDX mice treated with ruxolitinib, bortezomib, tofacitinib, and combinations compared with controls. P values were calculated by t test. ∗P < .05; ∗∗P < .01. (D) Bar plot showing the fold change in the spleen volume (measured by MRI) of RO2 and RO6 mice treated with ruxolitinib, bortezomib, tofacitinib, and combinations compared with controls. P values were calculated by t test. ∗P < .05; ∗∗P < .01. (E) Kaplan-Meier plot depicting overall survival of RO6 mice treated with vehicle, tofacitinib, ruxolitinib, or combination. P value was calculated using a log-rank test. (F) Line-dot plot showing changes in spleen volume (measured by MRI) of 3053 PDX mice treated with irinotecan (20 mg/kg per day) compared with controls in a timeframe of 40 days. P value was calculated by t test. ∗∗∗∗P < .0001. (G) Line-dot plot showing changes in percentage of T-ALL cells in blood of 3053 and 3119 PDX mice treated with irinotecan compared with controls in a timeframe of 80 days. Three of the 12 irinotecan-treated mice relapsed and died of leukemia. P value was calculated by t test. ∗∗∗∗P < .0001. (H) Kaplan-Meier plot depicting overall survival of 3053 and 3119 PDX mice treated with vehicle or irinotecan. Nine of the 12 irinotecan-treated mice died without leukemia. P value was calculated using a log-rank test. (I) Kaplan-Meier plot depicting overall survival of 8 different T-ALL PDX models (6467, 14741, 7072, 7155, 13002, 13356, 10512, and 5384) treated with vehicle or HDAC inhibitor panobinostat (n = 2 mice per arm, n = 32 mice total) (5 mg/kg per day). Mice belonging to 3 of the 8 models (5384, 7155, and 6467) died without leukemia. P value was calculated using a log-rank test. (J) Bar plot depicting the circulating T-ALL cells (difference between day 30 and day 0; day 0: first T-ALL detection in the blood) in 8 PDX models treated with vehicle or panobinostat. A single partially refractory model was identified (1 of 8, 7072 PDX T-ALL). P value was calculated by t test. ∗∗P < .001. (K) Bar plots depicting the percentage of circulating T-ALL cells in 4 PDX models (10512, 6467, 13002, and 13356) treated with vehicle, daunorubicin (1.2 mg/kg for 3 days/week), linsitinib (50 mg/kg per day), or combination (n = 2 mice per arm, n = 32 mice total). Mice treated with daunorubicin plus linsitinib displayed delayed leukemia progression compared with the other arms. P value was calculated by t test. ∗P < .05. (L) Kaplan-Meier plot depicting overall survival of 10512, 6467, 13002, and 13356 T-ALL PDX models treated with vehicle, daunorubicin, linsitinib, or combination. Mice treated with daunorubicin plus linsitinib survived until 3 months from enrollment. P value was calculated using a log-rank test.

This Feature Is Available To Subscribers Only