Nanobody-based retargeting of an oncolytic herpesvirus for eliminating CXCR4+ GBM cells: A proof of principle
Virotherapy
DOI:
10.1016/j.omto.2022.06.002
Publication Date:
2022-06-06T06:27:15Z
AUTHORS (13)
ABSTRACT
Glioblastoma (GBM) is the most aggressive primary brain tumor in adults, which remains difficult to cure. The very high recurrence rate has been partly attributed presence of GBM stem-like cells (GSCs) within tumors, have associated with elevated chemokine receptor 4 (CXCR4) expression. CXCR4 frequently overexpressed cancer tissues, including GBM, and usually correlates a poor prognosis. We created CXCR4-retargeted oncolytic herpesvirus (oHSV) by insertion an anti-human nanobody glycoprotein D attenuated HSV-1 (ΔICP34.5, ΔICP6, ΔICP47), thereby describing proof principle for use nanobodies target oHSVs toward specific cellular entities. Moreover, this virus armed transgene expressing soluble form TRAIL trigger apoptosis. In vitro, oHSV infects U87MG CXCR4+ patient-derived GSCs CXCR4-dependent manner and, when armed, triggers orthotopic xenograft mouse model, slows down growth significantly improves mice survival. Customizing diverse targeting multiple proteins appears as interesting approach tackling heterogeneity especially GSCs. Altogether, our study must be considered first step personalized virotherapies complement current treatments. IntroductionThe (CXCR4), described its role leukocyte trafficking or HIV infection,1Feng Y. Broder C.C. Kennedy P.E. Berger E.A. HIV-1 entry cofactor: functional cDNA cloning seven-transmembrane, G protein-coupled receptor.Science. 1996; 272: 872-877https://doi.org/10.1126/science.272.5263.872Crossref PubMed Scopus (3622) Google Scholar largely studied G-protein-coupled that activates various signaling pathways upon binding unique ligand CXCL12, also known stromal-cell-derived factor 1. overexpression reported wide range glioblastoma multiforme (GBM),2Alimohammadi M. Rahimi A. Faramarzi F. Alizadeh-Navaei R. 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Equine enters pathways, infection requires activation kinase ROCK1.J. 81: 10879-10889https://doi.org/10.1128/jvi.00504-07Crossref Campadelli Fiume), (MOI: 0.01, 0.1, Contrary oHSV/gD, led infectious foci J/C, no detected infection, demonstrating properly 2A). To evaluate cells, level (Figures S1A S1B) lentivirus CXCR4. ectopic flow cytometry S1B). then 0.1), real-time GFP imaging quantification Incucyte S3 2B, 2C, S2). As expected, replicated lines independently On contrary, remained only few infected, reflected weak EGFP difference non-infected clearly contrasted overtime oHSV/Nb-gD-infected confirming relies similar. curve U87MG-CXCR4+ No observed S3).Figure 2Efficacy retargetingShow full caption(A) Detargeting J1.1–2, (J1.1 HVEM+), nectin-1+) 72 h MOIs either wild-type (WT) (oHSV/gD) anti-hCXCR4 (oHSV/Nb-gD). Both pICP6, allowing visualization epifluorescence microscopy. Scale bars represent 5 mm. (B C) Retargeting (B) (C) Cells plated 96-well plates, (MOI 0.1) incubated analyses during h. confluency quantified every 6 Circles ratio green phase area mean ± SEM four wells. Statistical significance determined ordinary two-way ANOVA Bonferroni comparisons means pooled variance (ns, non-significant; ∗ p < 0.05, ∗∗ ∗∗∗∗ 0.0001). Images taken h, representative images hpi shown. Additional whole-well 24, 48, Figure S2. See S3View Large Image ViewerDownload Hi-res image Download (PPT)CXCR4-dependent oHSV/Nb-gDThe (T08, T013, T018, T033) directly established residual tissue obtained (Department Neurosurgery, CHU Liège, Belgium) maintained tumorospheres. opposition levels SOX2, POU3F2, SALL2 S4). percentage among analyzed 3A 3B ). While less than 3% T08 positive around 75% T033 receptor, T013 T018 intermediate. endogenous much lower 3B). compare non-retargeted cultured w
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