Fig. 3: 4-OI promotes selective replication of VSVΔ51 in colon tumor organoids. | Nature Communications

Fig. 3: 4-OI promotes selective replication of VSVΔ51 in colon tumor organoids.

From: Octyl itaconate enhances VSVΔ51 oncolytic virotherapy by multitarget inhibition of antiviral and inflammatory pathways

Fig. 3

a Confocal imaging of colon normal (NO) and tumor organoids (TO) pretreated with 4-OI (125 μM) for 24 h, then infected with VSVΔ51-RFP (1 × 106 pfu/well). Images taken two days post-infection using Hoescht dye as a DNA staining agent. Scale bars, 300 μm. b Flow cytometry counts of infected RFP-positive cells in enzymatically digested colon NO and TO organoids from patient 1 (P1) at two days post-infection with VSVΔ51-RFP (1 × 106 pfu/well), with or without 4-OI (125 μM). c qPCR analysis of VSV L gene expression normalized to a housekeeping gene (TBP) in colon NO and TO organoids from patient 1 (P1) at one-day post-infection with VSVΔ51 (1 × 106 pfu/well), with or without 4-OI (125 μM). d Imaging setup in a 48-well plate to evaluate the infectivity of VSVΔ51-RFP infection (1 × 106 pfu/well) solely and in combination with 4-OI in colon cancer organoids from 12 patients (P1 to P12) at 48 h post-infection. Organoid area evaluated either by GFP transduction or calcein green staining. Scale bars, 3000 μm. e, f Representative values from the imaging setup expressed as a percentage of RFP fluorescence from VSVΔ51 normalized to total GFP area from tumor organoids. g Remaining luciferase activity to assess in vitro cytotoxicity of VSVΔ51 alone or combined with 4-OI towards luciferase-expressing colon tumor organoids from different patients at 4 days of post-infection is presented as “survival”. Data are depicted as means ± SEM from two independent experiments performed in biological triplicates from one patient in (b), from one experiment performed in biological duplicates from one patient in (c), from one experiment performed in biological triplicates on tumoroids from 12 individual patients in (e, f), from one experiment performed in biological quadruplicates from 5 individual patients in (g). Pictures are from one representative experiment out of two in (a), and from one experiment performed on all individual patient material in (d). Statistics indicate significance by one-way ANOVA for (b, g), and two-tailed Student’s t-test for (e, f). Source data are provided as a Source Data file.

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