Synergy between VCR, DEX and ASP against ALL cell lines <i>in vitro</i>.

<p>Cell lines were exposed to VCR (open circles), DEX (open triangles), ASP (open squares), or the triple combination VXL (closed circles), at fixed ratios, and dose-responses were assessed using the DIMSCAN assay as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0033894#s4" target="_blank">Materials and methods</a>. Fractional survival of treated vs. untreated control cells is shown. Each condition included 12 replicates and error bars represent standard deviation. The data shown are representative of two independent experiments.</p

<i>In vivo</i> sensitivity of ALL xenografts to VXL and VXL/ABT-737 combination treatments.

<p>Female mice were engrafted with: ALL-2 (<b>A</b>); ALL-8 (<b>B</b>); ALL-10 (<b>C</b>); or ALL-17 (<b>D</b>) and treated with a diluent vehicle (controls, dashed black lines), ABT-737 (25 mg/kg, solid grey lines), VXL combination (solid black lines), or VXL+ABT-737 quadruple combination (dashed grey lines). Engraftment kinetics indicated by %huCD45⁺ cells in PB of individual mice (left panel) and Kaplan-Meier survival curves (EFS) (right panel) are shown. Shaded boxes represent the treatment period. All events were leukemia-related except for 1 and 4 in the VXL/ABT-737-treated group of the ALL-8, and ALL-10, respectively. In the ALL-17 quadruple drug combination cohort all mice were culled due to leukemia or toxicity unrelated morbidity.</p

LGD in response to VXL treatment in xenografts stratifies according to patient outcome.

<p>Median LGD obtained by VXL treatment for a panel of ALL xenografts derived from long term survivors (Alive) and from patients who died of their disease (DOD). There is evidence that the two groups are different (<i>p</i> = 0.0159) by Mann-Whitney two tailed test.</p

<i>In vivo</i> sensitivity of ALL xenografts to VXL and VXL/ATO combination treatments.

<p>Female mice were engrafted with: ALL-4 (<b>A</b>); ALL-7 (<b>B</b>); ALL-8 (<b>C</b>); or ALL-19 (<b>D</b>) and treated with a diluent vehicle (controls, dashed black lines), ATO (2.5 mg/kg, solid grey lines), VXL combination (solid black lines), or VXL+ATO quadruple combination (dashed grey lines). Engraftment kinetics indicated by %huCD45⁺ cells in PB of individual mice (left panel) and Kaplan-Meier survival curves (EFS) (right panel) are shown. Shaded boxes represent the treatment period.</p

<i>In vivo</i> sensitivity of ALL-19 to low dose VCR, DEX and ASP.

<p>Female mice were engrafted with ALL-19 cells and treated with vehicle (<b>A</b>); DEX (5 mg/kg) (<b>B</b>); VCR (0.15 mg/kg) (<b>C</b>); and ASP (1000 U/kg) (<b>D</b>); as single agents or the combination of the three drugs at the same doses (VXL) (<b>E</b>). The %huCD45⁺ cells in PB of individual mice (<b>A–E</b>); control vehicle-treated mice (dashed lines); drug-treated mice (solid lines). Kaplan-Meier analysis of EFS (<b>F</b>) control (grey solid line), VCR (grey dashed line), DEX (black dashed line), ASP (black dotted line), VXL (solid black line). All events were leukaemia-related. Shaded boxes represent the treatment period.</p

<i>In vivo</i> responses of ALL xenografts to ABT-737, VXL or VXL/ABT-737 combination treatments.

<p>Significant values are shown in bold.</p

Combination Indices of <i>in vitro</i> cytotoxicity assays.

<p>VCR, vincristine; DEX, dexamethasone; ASP, l-asparaginase.</p

Pharmacokinetic analysis of VCR, DEX and ASP in leukemias bearing NOD/SCID mice.

<p>Engrafted female mice (ALL-19) were treated with VCR (0.15 mg/kg), DEX (5 mg/kg), ASP (1000 U/kg) or their combination (VXL) at the same doses. Three mice each were sacrificed at specified time points and drug concentrations in plasma for VCR (<b>A</b>); DEX (<b>B</b>); and ASP (<b>C</b>) in the single agent or combination treatment were assessed as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0033894#s4" target="_blank">Materials and Methods</a>.</p

<i>In vivo</i> responses of ALL xenografts to ATO, VXL or VXL/ATO combination treatments.

<p>Significant values are shown in bold.</p

<i>In vivo</i> sensitivity of ALL xenografts to VXL combination treatment.

<p>Female mice were engrafted with: ALL-4 (<b>A</b>); ALL-7 (<b>B</b>); ALL-11 (<b>C</b>); or ALL-16 (<b>D</b>); and treated with a combination of VCR (0.15 mg/kg), DEX (5 mg/kg) and ASP (1000 U/kg). The %huCD45⁺ cells in PB of individual mice (left panel) and Kaplan-Meier analysis of EFS (right panel). Control vehicle-treated mice (dashed lines); drug-treated mice (solid lines). Shaded boxes represent the treatment period. No leukaemia related events were recorded for the drug treated group of ALL-16 engrafted mice.</p