Enhancement of lentiviral vectors gene delivery against VEGFR2 expressing cells by co-display of the binding and fusogenic moieties: a two molecules targeting approach.
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Abstract
Background: Pseudotyped lentiviral vectors (LVs) were used in cancer therapy as gene delivery systems for inhibiting tumor angiogenesis by targeting cells overexpressing "vascular endothelial growth factor receptor-2 (VEGFR2)". Herein, we report that switching from chimeric sindbis virus glycoprotein (SVG) harboring VEGFR2-specific nanobody (VEGFR2-Nb) to Two-Molecules Targeting Approach (TMTA: independent co-display of binding and fusogenic moieties) highly enhanced the transduction efficiency (TE) of the targeted LVs.
Methods: Several LVs co-displaying either the VEGFR2-Nb or the natural ligand (VEGF121) as targeting moiety, along with a de-targeted mutant form of SVG (as a binding deficient and fusion competent) fusogenic moiety were produced. LVs were constructed via various backbones and linkers (platelet-derived growth factor receptor "PDGFR" and CD28 as transmembrane domains, and HL and Fc as spacer domains).
Results: Expression and incorporation of the VEGFR2-Nb and SVG onto lentiviral particles were confirmed by flowcytometry and Western blotting while their co-display was demonstrated by virus-capture ELISA and virus-cell binding assays. LVs co-enveloped with fusogen and either VEGFR2-Nb or VEGF121 showed higher TEs in VEGFR2-expressing cells (72% and 91%, respectively) over LVs pseudotyped with chimeric fusogen containing the same nanobody (30%). In silico analyses indicated a direct correlation for the TE and the distance between the nanobody and the lipid bilayer.
Conclusion: Compared to the chimeric strategy, the two-molecule targeting approach of LVs, due to its flexible and modular nature provides higher TE and thus great potentials for targeted gene delivery.
期刊介绍：
Virology Journal is an open access, peer reviewed journal that considers articles on all aspects of virology, including research on the viruses of animals, plants and microbes. The journal welcomes basic research as well as pre-clinical and clinical studies of novel diagnostic tools, vaccines and anti-viral therapies.
The Editorial policy of Virology Journal is to publish all research which is assessed by peer reviewers to be a coherent and sound addition to the scientific literature, and puts less emphasis on interest levels or perceived impact.
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