Treatment schedule.

<p>Treatment schedule.</p

Effects of TL therapy combinations on NB tumor vascularization and perfusion.

<p>A. Representative T₂W fast SE images (top) and the corresponding ΔSco₂ (middle row) and ΔSo₂ (bottom) maps of control (left), TL-118^1/4 (middle column) and TL^1/4 + Gem (right) treated tumors (Bar = 1 cm). B. Mean ΔSo₂ values of the tumor and liver-ROI's calculated for control (blue), TL-118^1/4 (green) and TL^1/4 + Gem (purple) treated mice. C. Representative histological slides immuno-stained with the smooth muscle marker α-SMA (upper-rows) and with the endothelial cell marker CD31 (lower-rows), of control (top), TL-118^1/4 (middle) and TL^1/4 + Gem (bottom) treated tumors. Photographs were taken from the peripheral (left column) and central (right column) regions (original magnification ×20) demonstrating the higher vascularity in tumor periphery. D. Quantification of α-SMA positive vessels/HPF was determined from the tumor center (dark) and peripheral (light) areas for each of the treatment groups. (Mean ± SE; n = 5–7 mice/group).</p

TL-118 and Gemcitabine Drug Combination Display Therapeutic Efficacy in a MYCN Amplified Orthotopic Neuroblastoma Murine Model – Evaluation by MRI

<div><p>Neuroblastoma (NB) is the most common extra-cranial pediatric solid tumor with up to 50% of NB patients classified as having high-risk disease with poor long-term survival rates. The poor clinical outcome and aggressiveness of high-risk NB strongly correlates with enhanced angiogenesis, suggesting anti-angiogenic agents as attractive additions to the currently insufficient therapeutics. TL-118, a novel drug combination has been recently developed to inhibit tumor angiogenesis. In the current study, we used the SK-N-BE (2) cell line to generate orthotopic NB tumors in order to study the combinational therapeutic potential of TL-118 with either Gemcitabine (40 mg/kg; IP) or Retinoic acid (40 mg/kg; IP). We show that TL-118 treatment (n = 9) significantly inhibited tumor growth, increased cell apoptosis, reduced proliferation and extended mouse survival. Moreover, the reciprocal effect of TL-118 and Gemcitabine treatment (n = 10) demonstrated improved anti-tumor activity. The synergistic effect of these drugs in combination was more effective than either TL or Gemcitabine alone (n = 9), via significantly reduced cell proliferation (p<0.005), increased apoptosis (p<0.05) and significantly prolonged survival (2-fold; p<0.00001). To conclude, we demonstrate that the novel drug combination TL-118 has the ability to suppress the growth of an aggressive NB tumor. The promising results with TL-118 in this aggressive animal model may imply that this drug combination has therapeutic potential in the clinical setting.</p></div

Effects of B20 therapy on glioblastoma tumors.

<p>Tumor volume (mm³) for each individual tumor (<b>A</b>) and the average volume of the entire group (<b>B</b>), as a function of days post cell inoculation for control (black, n = 15) and B20 treated (blue, n = 6) mice. B20 showed only moderate effects on tumor growth kinetics. Comparison of the average diffusion (<b>C</b>) and HRI (<b>D</b>) values of tumors (blue; larger than 15 mm³) and the CLS (red) between control and B20 treated mice. Additionally, the HRI tumor values were also separated between borders (green) and the central area (purple). (<b>E</b>) Quantification of vessel count/HPF analyzed from 10 randomly selected HPF/tumor is based on the CD31 immunostaining. Tumor vascularity is significantly higher in the border (green) compared to the center (purple) in both groups. (<b>F</b>) Representative histological sections of control (left) and B20 treated (right) immunostained with Ki67 for proliferation (<b>top</b>) and TUNEL for apoptosis (<b>bottom</b>); Original magnification X40. *denotes statistical significance at P<0.05; **denotes statistical significance at P<0.01; ***denotes statistical significance at P<0.005.</p

Treatment effect on tumor growth and mouse survival.

<p>A. Tumor volume (mm³) for each individual mouse, as measured from T₂W MRI images as a function of days post cell inoculation in control (n = 19), Gemcitabine (Gem; n = 6), TL-118^1/4 (n = 9) and TL^1/4+ Gem combination (n = 10) treated mice. The dashed line indicates the maximal survival day of the control-treated mice. The b-values represent the average exponential coefficients of each treatment group. The b-values of all the treated groups (Gem, TL^1/4 and TL^1/4+ Gem) were significantly lower compared to control (p<0.0001). B. Representative T₂W anatomical axial images of Control, Gem, TL-118^1/4 and TL^1/4+ Gem treated tumors that were acquired on the indicated days (Bar = 1 cm) C. Kaplan-Meier survival analysis for each of the treated groups (*p<0.05; **P<0.0001; ***p<0.00001 compared to control). D. Box and Whisker plots of mean calculated b-values for each treated group (black square – median; * p<0.0001).</p

Effects of the different therapies on NB tumor proliferation and apoptosis.

<p>Representative histological sections of control (1^st column), TL-118^1/4 (2^nd column), Gem (3^rd column) and TL^1/4 + Gem (4^th column) treated mice. Slides were immuno-stained for proliferation (BrdU) (A) and apoptosis (TUNEL) (B); Quantification of BrdU positive (C) and TUNEL positive (D) cells/HPF analyzed from 10 randomly selected HPF/tumor ; n = 3–6 mice/group. Original magnification is indicated on the right image of each row. * p = 0.01, ** p<0.005. The histological sections were taken at the end of the experiments when the tumor load/mouse reached ethical limits.</p

TL-118^¼ drug composition.

<p>TL-118^¼ drug composition.</p

Glioblastoma tumor imaging characteristics.

<p>Representative T₂W image (<b>A</b>), T₁W image obtained before Gd-DTPA injection (<b>B</b>); contrast enhanced T₁W image obtained 1 min after Gd-DTPA injection (<b>C</b>); DWI map (<b>D</b>) and HRI map (<b>E</b>) with the corresponding H&E stained histological slide (<b>F</b>) of control tumor obtained 30 days post Gl-261 murine glioma cells inoculation. T-tumor, E-edema; bar-1 cm.</p

Glioblastoma imaging parameters progression over time.

<p>(<b>A</b>) Serial coronal FISP (first row) and T₂W (second row) images of a control tumor that were acquired on days 7, 14, 18 and 22. The corresponding DWI (third row) and HRI (fourth row) maps are also shown. ROIs delineating the tumor and contra-lateral healthy brain tissue are marked on the FISP and T₂W images (blue line). Increased HRI response during tumor progression can be seen both in the HRI maps and in the ROI-time-course (RTC) graphs (fifth row). The average diffusion values (<b>B</b>) and the HRI values (<b>C</b>) of the tumors (blue) and the contra-lateral side (CLS; red) are given as a function of time from cell inoculation. Note that the diffusion values are higher than the CLS throughout the entire period while tumor HRI increased over time. ***denotes statistical significance at P<0.005 compared to the CLS value.</p

HRI in the healthy brain.

<p>HRI maps were obtained without (<b>A, B, C, E, F, G</b>) or with (<b>I, J, K, M, N, O</b>) constant air flow above the head. HRI maps are presented for both positive and negative pixels (<b>A, I</b>) and the corresponding SI time course (<b>E, M</b>); only for positive pixels (<b>B, J</b>) and the corresponding SI time course (<b>F, N</b>) and only for negative pixels (<b>C, K</b>) and the corresponding SI time course (<b>G, O</b>). In the graphs the time-curse (TC, red line) is calculated only for active pixels in the brain ROI (marked in blue on the HRI maps) and the ROI time-course (RTC, blue line) is the TC multiplied by the ratio of the number of active pixels divided by the total number of pixels in the ROI. It can be seen that when airflow is used the negative response is significantly lower and randomly distributed (<b>K</b>) compared to maps acquired without airflow (<b>C</b>). The corresponding T₂W image is presented in (<b>D</b>) with the matching HRI superimposed on top (<b>H</b>). Note that the main blood vessels are clearly manifested on the HRI map. (<b>L</b>) The mean TC and RTC calculated from 44 HRI maps acquired with or without constant airflow are compared.</p