Journal: Scientific Reports

Article Title: The oncolytic adenovirus VCN-01 promotes anti-tumor effect in primitive neuroectodermal tumor models

doi: 10.1038/s41598-019-51014-1

Figure Lengend Snippet: Characterization of VCN-01 in PNET cell lines in vitro . ( A ) Expression of adenoviral receptors CAR, α v β 3 integrin and α v β 5 integrin in CNS-PNET cell lines PFSK-1 (left) and SK-PN-DW (right). Graph shows the percentage of stained cells for each receptor (Mean ± SD; n = 3). ( B ) PFSK-1 and SK-PN-DW (200,000 cells) infected with the GFP expressing vector AdTLRGDK at MOIs of 0, 0.1, 1, 10 or 100 PFU/cell. Graph indicates the percentage of GFP positive cells expression measured by flow cytometry at 24 h (white bars) or 48 h (black bars) after the infection (Mean ± SD; n = 3). ( C ) Detection of the viral proteins E1A and fiber in whole-cell lysates 48 h after being from PFSK-1 and SK-PN-DW from VCN-01 infected PFSK-1 and SK-PN-DW cultures. Grb2 was used as loading control protein. Blots from different parts of the same gel have been grouped to improve clarity of the image. ( D ) Viral titers in PFSK-1 and SK-PN-DW (50,000 cells/well) cultures 72 h after being infected with VCN-01 at MOIs 1 and 10. Bars represent total PFUs contained in the lysates (Mean ± SD; n = 3).

Article Snippet: 5 × 10 5 PFSK-1 or SK-PN-DW cells were engrafted into the caudate nucleus of 4–6 weeks female athymic nude (nu/nu) mice (Envigo, Huntingdon, UK) following the guide-screw system described by Lal et al . .

Techniques: In Vitro, Expressing, Staining, Infection, Plasmid Preparation, Flow Cytometry

Journal: Scientific Reports

Article Title: The oncolytic adenovirus VCN-01 promotes anti-tumor effect in primitive neuroectodermal tumor models

doi: 10.1038/s41598-019-51014-1

Figure Lengend Snippet: VCN-01 treatment results in cytotoxicity and immunogenic cell death. ( A ) Viability of PFSK-1 or SK-PN-DW cultures infected with VCN-01 at different MOIs ranging from 0 to 40 PFU/cell at 5 days post-infection. Graph indicates the percentage of viable cells relative to non-infected cultures, considered as 100% of viability (Mean ± SD; n = 3). ( B ) Measurement of DAMPs Hsp90α, HMGB1 and ATP in supernatants obtained from PFSK-1 and SK-PN-DW cultures three days after being infected with VCN-01 at their respective IC 50 (Mean ± SD; n = 3). ( C ) Representative images of calreticulin staining (red signal) in PFSK-1 (left) and SK-PN-DW (right) 4 h after being infected with either VCN-01 (bottom) or mock infected (top). Nuclei were stained in blue (DAPI), and green arrows indicate the location of calreticulin clusters in the cell membrane.

Article Snippet: 5 × 10 5 PFSK-1 or SK-PN-DW cells were engrafted into the caudate nucleus of 4–6 weeks female athymic nude (nu/nu) mice (Envigo, Huntingdon, UK) following the guide-screw system described by Lal et al . .

Techniques: Infection, Staining

Journal: Scientific Reports

Article Title: The oncolytic adenovirus VCN-01 promotes anti-tumor effect in primitive neuroectodermal tumor models

doi: 10.1038/s41598-019-51014-1

Figure Lengend Snippet: Anti-tumor effect of VCN-01 in PNET xenograft models. ( A,B ) Survival curves of mice bearing orthotopic PFSK-1 cells (A) or SK-PN-DW ( B ) tumors. Graphs represent overall survival (OS) with 95% confidence interval (blue and red shades) of mice treated with VCN-01 (blue line) or mock treated (red line). ( C – G ) Brain sections from mice bearing xenografts from either PFSK-1 ( C ) or SK-PN-DW ( D ) cells. Slices were stained with hematoxylin/eosin staining, or immunostained with anti-hexon (E, F) and hyaluronic acid binding protein. ( G , H ) Tumor regions (dotted square) have been magnified 10X in bottom pictures. VCN-1 (LT) (right column) corresponds to a long-term survivor. ( I ) Quantification of HA stained area (relative to total tumor area) in brain sections from mice bearing PFSK-1 tumors treated with either PBS or VCN-01 (Mean ± SD; n = 4/5).

Article Snippet: 5 × 10 5 PFSK-1 or SK-PN-DW cells were engrafted into the caudate nucleus of 4–6 weeks female athymic nude (nu/nu) mice (Envigo, Huntingdon, UK) following the guide-screw system described by Lal et al . .

Techniques: Staining, Binding Assay

Journal: Scientific Reports

Article Title: The oncolytic adenovirus VCN-01 promotes anti-tumor effect in primitive neuroectodermal tumor models

doi: 10.1038/s41598-019-51014-1

Figure Lengend Snippet: VCN-01 stimulate macrophage recruitment to the tumor periphery. ( A,B ) Brain sections from PBS and VCN-01 treated mice bearing xenografts from either PFSK-1 ( A ) or SK-PN-DW ( B ) cells immunostained with anti-F4/80 antibody. Tumor regions (dotted black squares) have been magnified in the middle panel, and labels T, TE and Br indicate tumor stroma, tumor edge and normal brain parenchyma, respectively. Red and green dotted squares have been amplified in the bottom panel to illustrate examples of amoeboid and ramified macrophages, respectively. ( C,D ) Quantification of F4/80 stained area (relative to total tumor area) in brain sections from mice bearing PFSK-1 ( C ) or SK-PN-DW ( D ) tumors treated with either PBS or VCN-01 (Mean ± SD). ( E–F ) Brain sections from PBS and VCN-01 treated mice bearing xenografts from either PFSK-1 ( E ) or SK-PN-DW ( F ) cells immunostained with anti-Iba1 antibody. ( G,H ) Quantification of Iba1 stained area (relative to total tumor area) in brain sections from mice bearing PFSK-1 ( G ) or SK-PN-DW ( H ) tumors treated with either PBS or VCN-01 (Mean ± SD).

Article Snippet: 5 × 10 5 PFSK-1 or SK-PN-DW cells were engrafted into the caudate nucleus of 4–6 weeks female athymic nude (nu/nu) mice (Envigo, Huntingdon, UK) following the guide-screw system described by Lal et al . .

Techniques: Amplification, Staining