Extended Data Fig. 10: Combination therapy with VTP-50469 and LEN results in additive anti-leukemic activity in vivo.
a, Immunohistochemistry for human CD45 antigen on bone marrow sections (sternum) on PDX/CBAM-68552 following 3 weeks treatment with vehicle, LEN 50 mg/kg daily, VTP-50469 0.1% rodent diet and the drug combination. b, Analysis of differentiation in circulating leukaemia cells using CD11b and CD14 expression assessed by flow cytometry in CBAM-68552 after 3 weeks drug treatment. Percentage of double-positive cells is indicated. Representative examples from individual mice are shown. c, Assessment of apoptosis in human CD45-positive cells from bone marrow of drug-treated mice using the CBSK-17D model following 2 weeks drug treatment. The percentage of total viable, human cells are indicated. Representative examples from individual mice are shown. d, Quantitation of apoptotic cells from bone marrow of mice transplanted with the CBSK-17D PDX model following 2 weeks drug treatment. Data represent mean+/−SEM with p-value by unpaired two-tail t-test. e, RNA-seq heatmap from PDX/CBAM-68552 following 3 weeks treatment with vehicle, LEN 50 mg/kg daily, VTP-50469 0.1% rodent diet and the drug combination. Heatmap displaying RNA-Seq DESeq2 statistical z-score for the co-regulated gene network between VTP-50469 and LEN f, RNA-seq heatmap displaying DESeq2 statistical z-score for genes detected in the ‘granulocyte’ pathway as additively deregulated using QIAGEN Ingenuity Pathway Analysis (IPA) analysis. g, Bar code plots using the gene set for HOXA9 regulated genes under each drug treatment from RNA-seq in vivo using the PDX/CBAM-68552 model. Normalized enrichment score and family-wise error rate p-value determined by the GSEA computational method. h, Gene set testing for reported MLL-fusion gene targets for LEN and VTP-50469. Family-wise p-value and normalized enrichment score (NES) determined by the GSEA computational method. i, IPA upstream regulator analysis. Graph displays selected activation z-scores for detected pathways with p-value <0.05. j, Measurement of peripheral blood circulating human CD45 positive cells from PDX mice transplanted with the DFAM-16835 PDX model (NPM1c) after two weeks of drug treatment in vivo. Data represent mean + /−SEM with p-value determined using unpaired, two-tail t-test.
