Dual-target anti-vascular endothelial growth factor/tumor necrosis factor-α nanobody precisely intervenes in...

This study aims to develop and evaluate a dual-target anti-VEGF/TNF-α nanobody (V5-3), exploring its dual role and mechanism in the treatment of Retinal neovascularization (RNV).

The V5-3 was first constructed and its effects on hypoxia-induced ARPE-19 cells and HUVEC cells were evaluated using Western blot, EdU assays, wound healing assays, among other methods. The role of V5-3 in suppressing angiogenesis and inflammation was further explored in a co-culture model designed to mimic RNV pathology.

The study found that V5-3 significantly inhibited TNF-α-induced apoptosis in L929 cells and suppressed proliferation, migration, and inflammatory cytokine expression of HUVEC cells stimulated by hVEGF₁₆₅, compared to the model group, with the most significant effect observed at high doses (all p < 0.01). Specifically, V5-3 effectively reduced angiogenesis through the VEGFR2 signaling pathway and significantly inhibited the expression of inflammatory cytokines in the NF-κB/TNF-α pathway by Western blot and RT-qPCR (all p < 0.01). Furthermore, V5-3 inhibited abnormal angiogenesis and inflammation in the RNV model under hypoxic conditions by reducing VEGFR2 phosphorylation and the activation of related downstream signaling molecules (all p < 0.01).

This study demonstrates that the V5-3, through dual-target inhibition of VEGF and TNF-α, shows more significant therapeutic effects in RNV compared to single-target antibodies.