Dual-Targeted Mcl-1 siRNA-Liposome Nanoparticles for Targeted Therapy of Allergic Rhinitis via CD4+CD44+ T Cell Modulation

To construct anti-CD4/anti-CD44 dual-modified Mcl-1 siRNA-loaded liposome nanoparticles (LPN) targeting allergic rhinitis (AR)-related CD4 + CD44 + T cells, and verify its in vitro Mcl-1 silencing and anti-inflammatory effects, in vivo therapeutic efficacy on ovalbumin (OVA)-induced mouse AR, as well as the underlying mechanism. The physicochemical and AR-related CD4 + CD44 + T cell targeting properties of dual-targeted LPN were systematically characterized. Its in vitro silencing efficiency on Mcl-1 and regulatory effect on Th2 polarization were detected in primary AR-related CD4 + CD44 + T cells. After nasal instillation of LPN in OVA-induced AR mice, therapeutic efficacy was evaluated by behavioral, histopathological and humoral indicators; the mechanism was explored by detecting Mcl-1 expression, inflammatory cell apoptosis and CD4 + IL-4 + CD44 + Th2 cell infiltration in nasal mucosa. The dual-targeted LPN had favorable physicochemical properties (138.6±12.3 nm, +21.5±3.1 mV), a high siRNA encapsulation efficiency (89.2±4.5%), 76.8% cumulative siRNA release at 48 h, and significantly higher targeting affinity than single/non-targeted LPN (P<0.001). It achieved a 72.5% Mcl-1 silencing efficiency in vitro and restored the Th1/Th2 immune balance. In vivo, it significantly alleviated AR behavioral symptoms, improved nasal mucosal pathology, reduced serum IgE levels and inflammatory cell infiltration (all P<0.001) by specifically silencing Mcl-1 in nasal mucosal CD4 + CD44 + T cells, promoting their apoptosis and reducing Th2 cell infiltration. The anti-CD4/anti-CD44 dual-modified Mcl-1 siRNA-LPN has excellent targeting ability and therapeutic efficacy for AR, which alleviates AR by regulating Mcl-1-mediated apoptosis of AR-related CD4 + CD44 + T cells, providing a novel precise siRNA nanotherapeutic strategy for AR. This graphical abstract depicts the development and therapeutic mechanism of anti-CD4/CD44 dual-targeted Mcl-1 siRNA-LPNs. These LPNs have high encapsulation efficiency, enable sustained siRNA release, and specifically target activated CD4 + CD44 + T cells. In vitro, LPNs efficiently silence Mcl-1, restore the Th1/Th2 cytokine balance and inhibit Th2 polarization. Nasal instillation in OVA-induced AR mice promotes inflammatory T cell apoptosis, reduces Th2 cell infiltration, markedly decreases serum IgE levels, and alleviates allergic rhinitis symptoms.