HA-modified lipid nanoparticles delivering ASO-Hotair attenuate macrophage-driven inflammation in sepsis-associated acute lung injury

Sepsis-associated acute lung injury (SA-ALI) is a life-threatening condition driven by macrophage-mediated inflammation and endothelial injury, yet effective targeted therapies remain lacking. ADAM17, a key sheddase regulating TNF-α release and NF-κB activation, amplifies inflammatory responses. Whether targeting HOTAIR can modulate ADAM17-driven macrophage inflammation and enable nanotherapy in SA-ALI remains unknown. We analyzed HOTAIR and ADAM17 expression in SA-ALI patients and CLP-induced SA-ALI mice. HA-modified MC3 lipid nanoparticles delivering Hotair-targeted antisense oligonucleotide (ASO-Hotair) were developed to evaluate targeting efficiency, safety, and therapeutic efficacy for SA-ALI. Mechanistic studies were performed in LPS-stimulated bone marrow-derived macrophages (BMDMs) and RAW264.7 macrophages and Hotair -knockout SA-ALI mice. HOTAIR and ADAM17 were significantly upregulated in SA-ALI patients and mice, and correlated with sepsis severity and inflammatory markers. HA-modified MC3-LNPs exhibited high encapsulation efficiency, stability, and biocompatibility, and enhanced uptake by pulmonary macrophages, with preferential lung accumulation in septic mice. MC3-LNPs-HA@ASO-Hotair effectively silenced Hotair, reduced ADAM17 levels, inhibited NF-κB activation, suppressed M1-like macrophage polarization in BMDMs and RAW264.7 macrophages, and attenuated macrophage-mediated endothelial injury in vitro. In vivo, treatment reduced macrophage infiltration, attenuated lung injury, decreased inflammatory mediators, and improved survival, with efficacy comparable to that of Hotair deficiency and no additional benefit in Hotair -knockout mice. Mechanistically, HOTAIR regulated ADAM17 expression, at least partly through miR-326, thereby promoting downstream NF-κB activation. Consistently, Hotair deficiency markedly reduced proinflammatory cytokines, chemokines, and endothelial injury markers, suppressed TNF-α/NF-κB signaling, alleviated lung injury, and improved survival in septic mice. Preferential delivery of MC3-LNPs-HA@ASO-Hotair to pulmonary macrophages enables Hotair silencing, attenuates ADAM17-driven macrophage inflammation, and improves survival in SA-ALI, providing a mechanistic and preclinical framework for lncRNA-targeted nanotherapy in sepsis.