Article Figures & Data
Figures
- Figure 1.
ABX displays greater bioavailability than CrEL-PTX and increased antiglioma effect in vivo. A, Comparison of IC50 of glioma cell lines across two common chemotherapeutic drugs: TMZ, temozolomide (n = 34), paclitaxel (n = 12). B–D, Biodistribution of ABX and CrEL-PTX. B, Plasma paclitaxel concentration at 45 and 180 minutes (*, P = 0.0285; **, P = 0.0056). C, Ratio of brain to plasma paclitaxel concentration at 45 minutes (ABX n = 9, CrEL-PTX n = 7, P = 0.0193) and 180 minutes (ABX n = 4, CrEL-PTX n = 3, P = 0.0349). D, Ratio of heart or liver to plasma PTX concentration at 45 minutes (ABX n = 9, CrEL-PTX n = 7, P < 0.0001). Data plotted, mean ± SD. E, In vivo antiglioma effect of ABX: MES83 cells were used to establish orthotropic xenografts and groups of 12 mice were randomized to treatment groups as indicated. Survival for each is plotted in Kaplan–Meier graphs and evaluated through log-rank test. Arrows represent treatment days (*, P = 0.0423; **, P = 0.0041).
- Figure 2.
Sodium fluorescein is a visual marker for ultrasound (US) BBB disruption. A, Fluorescent imaging: mice injected intravenously NaFL were treated with ultrasound and compared with nonsonicated controls. Brains were harvested and imaged using Nikon AZ100 microscope at 4× magnification with FITC filter cube (left) and SII Lago in vivo Imaging system (ex/em 465/530nm). B, NaFl and paclitaxel correlation: 16 brain samples from sonicated and nonsonicated mice were analyzed for NaFl and paclitaxel concentration through LC/MS. Correlation was determined by calculating Pearson coefficient (r = 0.8987, P < 0.0001).
- Figure 3.
Ultrasound increases ABX and CrEL-PTX brain penetration. A, Representative image of how samples were dissected. B–D, Following sonication and NaFl administration, mice were injected with either ABX or CrEL-PTX at 12 mg/kg. Paclitaxel concentration was determined in fluorescent, nonfluorescent and nonsonicated control samples through LC/MS. Data plotted are mean ± SD. Significance was determined one-way ANOVA test. Brain/plasma paclitaxel 45 minutes after sonication (***, P = 0.0002, ns; B) and brain/plasma paclitaxel 180 minutes after sonication (*, P = 0.0241; **, P = 0.0017; C). D, Absolute concentration of paclitaxel in US+ABX-treated mice compared with human glioma cell line paclitaxel IC50 concentration from Sanger/CCLE database (***, P = 0.0006, ns).
- Figure 4.
Ultrasound-delivered ABX differs in therapeutic profile between two patient-derived xenograft models. A, Cell viability: GBM12 and MES83 short-term explant cultures were exposed to increasing doses of ABX and viability after 72 hours was determined by CellTiterGlo. Dose–response curves represent three replicates. B, Experimental timeline. C, Five days after tumor implantation, mice were randomized to treatment groups as indicated and survival is plotted through Kaplan–Meier graphs. Survival differences were determined through log-rank analysis. Mice that did not die due to tumor burden were censored from this analysis. Censored subjects are denoted by tick mark on the day they were removed from the study. (MES83: **, P = 0.0041; **, P = 0.0036; ***, P = 0.0006; GBM12: ****, P < 0.0001; ns (P = 0.2590); ****, P < 0.0001). D, H&E stain of tumor histology from untreated control mice. Left, MES83 xenograft. Right, GBM12 xenograft. White arrows, blood vessels. B, Brain tissue; T, Tumor mass. White scale bar, 50 μm.
- Figure 5.
Toxicity evaluation of ultrasound-delivered paclitaxel therapy. A, Bodyweight of mice following single course of treatment; 10 mice were evaluated for each treatment condition. Data plotted are mean ± SD. B, Representative photomicrographs of H&E-stained axial brain sections from mice following multiple courses of ultrasound-delivered paclitaxel. Damage was most severe in ultrasound-delivered CrEL-PTX (top left, arrowhead, and bottom left), although ultrasound-delivered CrEL alone also elicited marked white matter damage (top central, arrowheads, and bottom central). Ultrasound-delivered ABX showed only small patches of damage to deep white matter tracts in some cases (top and bottom right, arrowheads). Scale bar, 200 μm in bottom panels and 800 μm in top panels.
Tables
- Table 1.
Toxicity evaluation from multiple courses of US-delivered chemotherapy.
Nontumor-bearing mice i.c. glioma PDX modela Treatment Mortality Signs of CNS Pathology Mortality Signs of CNS pathology Total survival (%) Ultrasound only 0/8 2/4 Small focal white matter vacuolation (WMV), Hemosiderin-laden macrophages (HLM) 0/19 27/27 (100%) Ultrasound + Cremophor EL PTX (12 mg/kg) 4/7 3/4 severe necrosis, hemorrhage, Diffuse axonal injury (DAI), hippocampal damage 3/7 (42%) Ultrasound + Cremophor EL (5% in Saline) 3/8 1/3 DAI 2/3 small focal WMV 5/8 (62.5%) Ultrasound + Abraxane (12 mg/kg) 1/6 3/4 Focal WMV 0/20 25/26 (96%) Ultrasound + Abraxane (24 mg/kg) 0/5 1/3 HLM 5/21 1/3 cytotoxic edema 1/3 DAI 1/3 focal WMV 21/26 (81%) Ultrasound + Abraxaneb (24 mg/kg) 0/10 10/10 (100%)
Supplementary Data
- Supplementary Materials and Data - Supplementary Methods and Materials Figures S1: Schematic representation of LIPU generator set up. Supp Table 1: Summary of histological evaluation of CNS after one course of US-PTX Supp. Table 2: STR results of PDX cell lines. Figures S2: Cell viability in response to PTX and ABX
Volume 26, Issue 2
