Fig. 6: Susceptibility of cancer cell lines to H-1PV oncolysis is directly correlated with LAMC1 expression levels.

We tested 53 cancer cell lines from the NCI-60 panel for sensitivity to H-1PV infection using the xCELLigence Real-Time Cell Analyser (ACEA Biosciences, Inc). Cancer cell lines were left untreated or infected with the indicated H-1PV multiplicity of infection (MOI; expressed as plaque forming units (pfu) per cell). Cell growth was monitored in real time for a total of 168 h and expressed as normalised cell index (CI). a Kinetic response profiles of one H-1PV-sensitive (SNB-75) and one H-1PV-resistant (COLO205) cancer cell line are shown as representative examples; at least n = 3 biologically independent samples; error bar for the curve represents the mean values ± SD. b The CI values obtained by xCELLigence analysis were used to calculate the EC50 values (viral MOI that kills 50% of the cell population) at four time points after infection (24, 48, 72 and 96 h; see also Supplementary Fig. 12). Based on the EC50 value corresponding to 72 h, cancer cell lines were classified into the six indicated groups. c Scatterplot. Single points indicate the LAMC1 expression levels in the 53 cancer cell lines that were screened for their susceptibility to H-1PV induced oncolysis (expressed as EC50). Source for the LAMC1 expression levels is the NCI-60 COMPARE database. d LAMC1 expression is robustly anticorrelated with EC50 value. Gene expression data from the NCI-60 (53 cell lines) and the CCLE (38 cell lines) panels were correlated with EC50 values (Pearson correlations). Bar plot shows the correlations, r, obtained in each dataset independently together with P values (both with significant correlations at P < 1E-3, null hypothesis: r = 0). These were generated in R (ggplot2 library). e LAMC1 mRNA expression levels correlate with H-1PV efficacy of inducing cell lysis in glioblastoma cell lines. The indicated glioma-derived cell lines were infected with H-1PV (MOI 10 pfu/cell) for 72 h before being processed for an LDH assay to measure H-1PV-induced cell lysis (left panel). The independent experiment shown is repeated twice; n = 3 biologically independent samples. The total RNA from these six glioblastoma cell lines and astrocytes was isolated, and LAMC1 mRNA expression levels were measured using NanoString analysis (right panel). Numbers on the top of the columns indicate gene expression fold changes between highly H-1PV-sensitive (NCH125 or NCH37) and poorly susceptible (U251, LN308, T98G, and A172-MG) cancer cell lines. The independent experiment is shown; n = 1 (NCH125, NCH37, U251 and A172-MG); n = 2 biologically independent samples (LN308 and T98G); n = 3 biologically independent samples (Astrocytes). f LAMC1 mRNA expression levels correlate with both NS1 mRNA expression levels and the transduction efficacy of H-1PV. The total RNA from untreated (mock) or recH-1PV-EGFP-infected P3 and P13 GBM 3D spheroids was isolated, and LAMC1 (left panel) and NS1 (middle panel) mRNA expression levels were measured using NanoString analysis. A representative image of H-1PV transduction in P3 and P13 spheroids is shown in the right panel. For left panel the independent experiment is shown; n = 2 biologically independent samples for P3 and P13 mock treated whereas n = 1 for P3 and P13 infected with recH-1PV-EGFP. For middle panel, the independent experiment is shown; n = 1. For right panel, a single experiment with 10 replicates was carried out (n = 10). The transduction efficacy was analysed in these spheroids using the IncuCyte ZOOM™ live cell imaging system. The scale bar corresponds to 100 μm. Error bars for all data indicate the mean values ± SD. Source data are provided as Source Data file.