Abstract 5009: Translational efficacy of oncolytic HSV-1 in glioblastoma using a human autologousex vivoplatform, CANscript™

Agent-based computational modeling of glioblastoma predicts that stromal density is central to oncolytic virus efficacy

“…From our calculations ( Smalley et al., 2019 ), there were on average approximately 9740 glioblastoma cells in a tumor slice. However, tumor density varied according to different experimental conditions, and we found a high degree of heterogeneity in the immune cell subset counts across the slices.…”

“…Glioblastoma tissue (n = 4 patient samples) was cut manually into slices or fragments of various sizes up to 1 mm thick, and treated with rQNestin (0.1–5 PFU per cell, for 24 h) by adding the virus in a 100 L drop of saline to the piece of tissue submerged in 400 L of complete medium and incubated at 37 with 5% CO2 ( Smalley et al., 2020 ). Using a “sequential imaging strategy”, which we described recently ( Smalley et al., 2019 ), serial sections were then fixed and either stained with hematoxylin and eosin (H&E), or immunostained for HSV-1 (DAKO, B0114), GFP (GFP antibody CAT# AM1009a from Abgent, clone 168AT1211), or cleaved caspase-3 (Cell signaling Tech, for clone information see Goldman et al., 2015 ). The impact of rQNestin on the spatial context and activity of the immune compartment in the glioblastoma ex vivo slices was further assessed by multiplex immunohistochemistry (mIHC).…”

“…The dimensions of our glioblastoma tumour tissue slice simulation were calibrated to those of an average ex vivo glioblastoma tumour slice from CANscript cultures containing tumour cells, stroma, and tumour infiltrating immune cells from patient tissue biopsies and resections ( Smalley et al., 2019 ) ( Figure 1 , Main Text). Tissue slices were collected from these samples at the end of the experiment by slicing the cultured tumour into deep tumour tissues ( Figure S10 A).…”

“…We conducted a series of control experiments where CANscript ex vivo tissue samples were grown for 0, 24, 48 or 72 h and their band area calculated using HALO ( Smalley et al., 2019 ). Two tumour slices from each patient (1098 and 1167) were analyzed with HALO at 0 h and the area of each band was calculated as .…”