Supplemental Figure S 1 A B Supplemental Figure

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Supplemental Figure S 1 A B Supplemental Figure 1: Specificity of VU 661013 for

Supplemental Figure S 1 A B Supplemental Figure 1: Specificity of VU 661013 for MCL-1. A, Binding affinity of VU 661013 toward purified human MCL-1, BCL-2, and BCL x. L in biochemical assays. B, Correlation of MCL-1 protein content to growth inhibiton in response to suprahigh dosing of VU 661013 in insensitive AML cell lines.

Supplemental Figure S 2 A B C D E F Supplemental Figure 2: Growth

Supplemental Figure S 2 A B C D E F Supplemental Figure 2: Growth inhibition with VU 661013 does not correlate well with protein Levels. A-F, Ratio of BCL-2, BCL-x. L and MCL-1 protein levels against AML cell line sensitivity to an MCL-1 inhibitor.

A B Supplemental Figure S 3 C Supplemental Figure S 3: Specificity of VU

A B Supplemental Figure S 3 C Supplemental Figure S 3: Specificity of VU 661013 for human MCL-1. A Binding affinity of VU 661013 toward purified human and mouse Mcl-1 in a biochemical assay. B Species specificity and PK information for VU 661013. C Hemotoxylin and Eosin staining of spleen, kidney, liver and heart tissue in vehicle and VU 661013 75 mg/kg treated NSGS mice after 21 days of daily dosing, 20 x magnification.

Supplemental Figure S 4 A Supplemental Figure S 4: Leukemia chimerism in setting of

Supplemental Figure S 4 A Supplemental Figure S 4: Leukemia chimerism in setting of continued treatment with MCL-1 inhibitor in cell line xenograft. Mice engrafted with MV-4 -11 cells were treated with VU 661013 for 4 -6 weeks and human CD 45+CD 33+ cells were measured in the blood, bone marrow and spleen. Vehicle day 28 (n= 4), VU 661013 75 mg/kg day 28 (n= 5) and VU 661013 75 mg/kg day 42 (n= 4).

Supplemental Figure S 5 A B C D

Supplemental Figure S 5 A B C D

Supplemental Figure S 5: Combination treatment with VU 661013 and VEN in human cell

Supplemental Figure S 5: Combination treatment with VU 661013 and VEN in human cell lines. A, VU 661013 and VEN combination growth inhibition dose curves. B, Optimal dosing to maximize growth inhibition differed between cell lines and survival fraction of treated cells compared to DMSO control plotted over concentration of drug. C, Combination index reveals synergy in cell lines with combination VU 661013 and VEN. Points represent the estimated combination index (CI), CI < 1 represents synergy, CI=1 represents additivity, and CI > 1 represents antagonism. Upper 95% confidence intervals that exclude 1 indicate statistically significant synergistic effects. (two-sided p<0. 05). Synergy data shown is representative of 3 individual experiments with 95% confidence interval. D, Other cell lines maintained a resistant phenotype.

A Supplemental Figure S 6 B D C E

A Supplemental Figure S 6 B D C E

Supplemental Figure S 6: Specificity of combination treatment in MCL-1 dependent PDX. A, Ex

Supplemental Figure S 6: Specificity of combination treatment in MCL-1 dependent PDX. A, Ex vivo analysis from AML 002, data from bone marrow harvested from mouse (mean SEM). Points represent the estimated combination index (CI), CI < 1 represents synergy, CI=1 represents additivity, and CI > 1 represents antagonism. Upper 95% confidence intervals that exclude 1 indicate statistically significant synergistic effects. (two-sided p<0. 05) Synergy data shown is from two biological replicates with a 95% confidence interval, and is representative of 2 individual experiments. B, Combination treatment with VU 661013 and VEN in PDX models and effects on leukemia-associated splenomegaly. C, Hemotoxylin and Eosin staining of spleen, kidney, liver and heart tissue in vehicle and VU 661013 75 mg/kg + VEN 15 mg/kg treated NSGS mice after 21 days of daily dosing. 20 x magnification. D,

Supplemental Table S 1 Supplemental Table 1: Characteristics of AML patient samples and normal

Supplemental Table S 1 Supplemental Table 1: Characteristics of AML patient samples and normal human bone marrow tested with VEN and VU 663013.