Fig. 6

Targeting ADAM10 inhibits homing, reduces LSC frequency and sensitizes to chemotherapy in vivo. A Workflow for early engraftment assay. B Quantification of PDX cells homing to the BM. Number of DMSO- (GI group: n = 6, Aderbasib group: n = 5) or inhibitor- (GI: n = 8, Aderbasib 10 µM: n = 5) treated ALL-199 or DMSO- (n = 5) or inhibitor- (n = 5) treated ALL-265 were analyzed. Data were normalized to the mean of the respective DMSO group. Each dot represents one mouse. **** p < 0.0001, *** p < 0.001, *p < 0.05 by paired t-test. C Quantification of the limiting dilution transplantation assay. Mean (solid lines) and 95% confidence interval (CI, dashed line) are depicted (ALL-199 n = 25). Bar graph depicts relative LIC frequency of ADAM10 KO cells normalized to control. D Quantification of colony-forming unit assay with PDX AML-356 or AML-388 cells with or without ADAM10 KO. Each dot represents one replicate. *p < 0.05 by paired t-test. E Quantification of colony-forming unit assay with PDX AML-356 or AML-388 cells treated with the ADAM10 inhibitor GI254023X (GI, 100 µM) or DMSO for 72 h. Each dot represents one replicate. * p < 0.05 by paired t-test. F Quantification of colony-forming unit assay with healthy human CD34 + blood progenitor cells treated with the ADAM10 inhibitor GI254023X (GI, 100 µM), Aderbasib (AD, 10 µM) or DMSO for 72 h. Each dot represents one replicate. ns by Holm-Sidak’s multiple comparisons test vs. DMSO. G Workflow of the competitive in vivo chemotherapy trial. ADAM10 KO cells marked with mTagBFP or CTRL cells marked with T-Sapphire were injected into groups of mice in a 4:1 ratio of ADAM10 KO:CTRL cells to compensate for the disadvantage of ADAM10 KO cells. Tumor growth was monitored by repetitive bioluminescence in vivo imaging. Mice were sacrificed at start of therapy (SOT) or following treatment with chemotherapy or PBS. Percentages of the KO and CTRL populations among the isolated human cells (ADAM10 KO + CTRL cells) were determined by flow cytometry. H Representative in vivo bioluminescence imaging pictures of mice carrying AML-661 PDX cells treated with cytarabine (AraC, n=3, loss of 2 mice due to drug-related toxicities) or PBS (n=4, 1 mouse was removed as extreme value) at the indicated time points. I Quantification of all images taken from mice carrying AML-661 PDX cells over time. J Quantification of human cells in the BM in AML-661. *** p < 0.001, by unpaired t-test. K Quantification of distribution of ADAM10 KO PDX cells in AML-661 at injection, start of therapy (SOT) and after treatment with PBS or AraC. **** p < 0.0001, *** p < 0.001 by unpaired t-test. L In vivo chemotherapy trial with ALL-265 treated with vincristine (VCR, n = 3), Cyclophosphamide (Cyclo, n = 3) or PBS (i.p., n = 4). Quantification of distribution of ADAM10 KO PDX cells in ALL-265 at injection and after treatment with PBS, VCR or Cyclo. *** p < 0.001, * p < 0.05 by unpaired t-test