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Background <p>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.</p> Methods <p>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).</p> Results <p>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.</p> Conclusion <p>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.</p>

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Enhancement of lentiviral vectors gene delivery against VEGFR2 expressing cells by co-display of the binding and fusogenic moieties: a two molecules targeting approach

  • 期刊论文

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  • 发表日期：

    2025年9月24日

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  • Roshanak Ahani,
  • Mohammad Hossein Etemadzadeh,
  • Nasir Mohajel,
  • Mahdi Behdani,
  • Reza Ahangari Cohan,
  • Mohamad Reza Kalani,
  • Navid Madani,
  • Farzin Roohvand,
  • Kayhan Azadmanesh

摘要

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.