Abstract

Biologics have emerged as frontline therapies for uveitis due to their targeted immunomodulatory effects. However, conventional monoclonal antibodies remain confined to single-target engagement. In addition, intravitreal injections carry risks of complications associated with repeated administration, whereas noninvasive delivery is hindered by compromised bioavailability across the complex ocular barriers. Together, these limitations restrict the effectiveness of current therapeutic approaches. Herein, bioinformatic screening identified VEGF and tumor necrosis factor alpha (TNF-α) as core pathogenic mediators in uveitis. Subsequently, we engineered a VEGF/TNF-α bispecific antibody (V5-3) and assembled it with a dithiolane molecule (DM) into nanoparticles (V5-3-NPs), thereby creating a noninvasive topical nanoformulation for uveitis therapy. Compared with V5-3, V5-3-NPs significantly enhanced cellular uptake, inhibited microglial proliferation and migration, and reduced the release of pro-inflammatory cytokines. In both in vitro and in vivo models, the synergistic bispecific targeting of V5-3-NPs enhanced anti-inflammatory and anti-angiogenic efficacy, thereby effectively alleviating clinical manifestations and histopathological damage. The negligible toxicity observed across models confirmed its safety profile and supported clinical feasibility. This work tackles key unmet needs in uveitis therapy by overcoming inefficient drug delivery and single-target constraints. By integrating bispecific targeting with nanocarrier-based delivery, it offers a distinct technological advance and a clinically viable strategy for uveitis treatment.