Supplemental Figures S1-S7 from Efficacy of the MDM2 Inhibitor SAR405838 in Glioblastoma Is Limited by Poor Distribution Across the Blood–Brain Barrier
journal contribution
posted on 2023-04-03, 15:05 authored by Minjee Kim, Daniel J. Ma, David Calligaris, Shuangling Zhang, Ryan W. Feathers, Rachael A. Vaubel, Isabelle Meaux, Ann C. Mladek, Karen E. Parrish, Fang Jin, Cedric Barriere, Laurent Debussche, James Watters, Shulan Tian, Paul A Decker, Jeanette E. Eckel-Passow, Gaspar J. Kitange, Aaron J. Johnson, Ian F. Parney, Panos Z. Anastasiadis, Nathalie Y.R. Agar, William F. Elmquist, Jann N. Sarkaria<p>Fig. S1: MDM2 transcript expression. RNA extracted from flank tumor xenografts from 51 PDX models was assessed by qRT-PCR for MDM2 expression in pooled triplicate samples and compared to expression in normal brain (NB). Fig. S2: MDM2 Amplification in PDX lines. Copy-number profiles were generated from whole exome sequencing of flank xenografts. MDM2 exons are highlighted in red. GBM10, GBM12, and GBM102 show normal MDM2 copy number. GBM108 and GBM143 demonstrate high-level amplification of MDM2. Fig. S3: Apoptosis by TUNEL assay in GBM108. Established flank tumor xenografts were randomized and treated for 5 days with placebo (n=5 mice) versus 50 mg/kg SAR405838 (n=5 mice). Tumor samples were then processed for TUNEL staining. TUNEL-positive cells were quantitated in six high-powered fields (HPF) for each sample. Results are plotted as mean and standard error of the mean. * P<0.05, ** P<0.01, *** P<0.001. Fig. S4: Blood brain barrier integrity in orthotopic tumors. Near-moribund mice with orthotopic GBM108 tumors were injected with TexasRed-3 kDa dextran conjugate 10 min before euthanasia and processed for cresyl violet and fluorescent microscopy on serial histology sections. Accumulation of TR-dextran within the tumor reflects disruption of the BBB. Results presented are representative of five mice analyzed. Scale bar = 500 µm. Fig. S5: VEGFA expression in GBM108 models. Short term explant cultures from parental GBM108, GBM108-empty vector (EV), and GBM108VEGFA were processed for an ELISA to measure VEGFA levels. One-way ANOVA. * P<0.05, ** P<0.01, *** P<0.001. Fig. S6: Effects of VEGFA expression on SAR405838 sensitivity. Short-term explant cultures from GBM108-Vector and GBM108-VEGFA were treated with indicated concentrations of SAR405838 for 24 hours and then relative cell number analyzed in a CyQuant assay. Fig. S7: Spatial distribution of SAR405838 in GBM108-Vector and GBM108-VEGFA intracranial tumors by MALDI-MSI.</p>
