Inhibition of shedding of ADAM17 substrates by D1(A12) and infiximab.

<p>Concentrations of the cleaved products of ADAM17 substrates in plasma and ascites fluid at the endpoint (day 33), as measured by ELISA. Horizontal bars represent the mean and standard deviation. Asterisks show the groups significantly different (P<0.05) from the PBS control group in the same fluid type.</p

Distribution of D1(A12) and infliximab IgG <i>in vivo</i> at the end of the efficacy study.

<p>Samples were taken at endpoint (day 33), 20 hours after the final antibody dose. (A) Concentrations in plasma and ascites fluid. Horizontal bars represent the mean and standard deviation. (B) Concentrations in homogenates of tissue from tumour in pancreas and omentum. (C) Immunohistochemistry to detect human IgG in IGROV1-Luc tumour and liver. Top: mouse ID 901, treated with D1(A12), with an arrow marking the position of a blood vessel; middle: mouse ID 903, treated with infliximab; bottom: mouse ID 902, treated with vehicle. Images were taken at 20X magnification.</p

Inhibition of IGROV1-Luc tumour growth <i>in vivo</i> by D1(A12) IgG.

<p>Growth of IGROV1-Luc i.p. xenografts in mice dosed weekly with vehicle, 10 mg/kg infliximab or 10 mg/kg D1(A12), measured by bioluminescence. The mean and standard deviation of day 32 tumour burden is shown, expressed as Avg Radiance. N = 12 for vehicle, n = 8 for infliximab and N = 11 for D1(A12).</p

<i>In vitro</i> activity of D1(A12) antibody.

<p>(A) D1(A12) IgG inhibits PMA-induced shedding of ADAM17 substrates into IGROV1-Luc cell culture medium. Medium was collected 90 minutes after addition of PMA (100 ng/ml), D1(A12) IgG (200 nM) or solvent control. The proteins were quantified by ELISA. (B) Dose-dependent inhibition of TNF- α shedding by 1 hour pretreatment with D1(A12), N-TIMP-3 or control human plasma IgG prior to PMA stimulation. (C) D1(A12) IgG inhibits constitutive shedding of TNF-α from IGROV1-Luc cells into culture medium. Medium was collected after 48 hours of incubation with or without IgGs at 200 nM. Error bars show the standard deviation.</p

Simulation in CYCLOPS of cell cycle and ligand modulation.

<p>EGF stimulation of malignant <b>(a)</b> and normal cells <b>(b)</b>. “Proportion cells” correspond to the increase in number of cells i.e. 10 = 10 times more cells.</p

Diagram of global processes and signalling pathways modelled in CYCLOPS.

<p><b>(a)</b> Cell cycle partition, checkpoint and apoptosis process. <b>(b)</b> Signalling processes modelled in the G1-S and <b>(c)</b> spindle assembly checkpoints. <b>(d)</b> Signalling modelled in the MAP kinase and <b>(e)</b> apoptosis pathways.</p

CYCLOPS simulations of actinomycin D and paclitaxel combinations.

<p><b>(a)</b> Actinomycin D dose-response are shown for malignant (left) and normal (right) cells. <b>(b)</b> Combination dose-response for actinomycin D+paclitaxel for simultaneous administration (24 hours) and <b>(c)</b> when delaying actinomycin D by 12 hours (malignant left, normal right). Arrows highlight magnitude of antagonistic effect when adding paclitaxel to 30nM actinomycin D.</p

Simulations of palbociclib and gemcitabine effects.

<p><b>(a)</b> palbociclib effects on malignant (left) and normal (right) at several concentrations (“proportion cells” correspond to the increase in number of cells i.e. 10 = 10 times more cells). <b>(b)</b> Modulation of the cell cycle distribution (left malignant, right normal, arrows indicate changes with increasing concentrations). <b>(c)</b> gemcitabine effects on malignant (left) and normal (right) at several concentrations. <b>(d)</b> Simulated dose-response surface for the palbociclib+gemcitabine combination. The arrow shows the rise of antagonistic effect with normal cells when increasing palbociclib concentration. <b>(e)</b> gemcitabine delayed administration protocol. (f) Effects of varying the time delay on normal (blue line, shown as % control) and malignant cells (red line). The ratio of normal to malignant is shown in green.</p

Properties of the cell lines modelled.

<p>Most values were from the ATCC website.[<a href="http://www.ploscompbiol.org/article/info:doi/10.1371/journal.pcbi.1005529#pcbi.1005529.ref055" target="_blank">55</a>] It should be noted that the cytokinetic properties of cell lines vary substantially according to culture medium, concentration of serum and growth factors, inoculum density and oxygen and CO₂ concentration. The values shown are the ones used in CYCLOPS and may be regarded as typical values for early-stage cultures at low cell density under standard levels of O2 and CO₂.</p

Paclitaxel modulation of the cell cycle distribution.

<p>24hours paclitaxel treatment was simulated and the resulting cell cycle distribution is shown at 12, 24 and 27 hours (3 hours after wash-out) for <b>(a)</b> normal and <b>(b)</b> malignant cells. Plain bars show the reference cell cycle distribution prior to treatment while broken-line bars show the distribution evolution over time.</p