Lung-Targeted Lipid Nanoparticles Delivery of Wogonin for Pulmonary Fibrosis in Mice via Modulation of Cellular Proteostasis

Libo Wang,1,2,* Fei Lin,1,* Fangli Gao,2,* Zhichao Jia,2 Junwei Liu,3 Xu Liu,4 Jie Shang,5 Xiangli Ru,2 Yilin Zhao,6 Tianhao Zhao,2 Lin Yang,2 Yuming Guo,2 Min Zhang7 1Life Science Research Center, The First Affiliated Hospital of Xinxiang Medical University, Xinxiang, Henan, People’s Republic of China; 2School of Chemistry and Chemical Engineering, Henan Normal University, Xinxiang, Henan, People’s Republic of China; 3School of Pharmacy, Xinxiang Medical University, Xinxiang, Henan, People’s Republic of China; 4Department of Anesthesia and Perioperative Medicine, The First Affiliated Hospital of Xinxiang Medical University, Xinxiang, Henan, People’s Republic of China; 5Department of Pathology, the First Affiliated Hospital of Xinxiang Medical University, Xinxiang, Henan, People’s Republic of China; 6Department of Cardiology, The Third Affiliated Hospital of Xinxiang Medical University, Xinxiang, Henan, People’s Republic of China; 7King’s College London British Heart Foundation Centre of Research Excellence, School of Cardiovascular and Metabolic Medicine & Sciences, London, UK*These authors contributed equally to this workCorrespondence: Yuming Guo, School of Chemistry and Chemical Engineering, Henan Normal University, Xinxiang, Henan, 453007, People’s Republic of China, Email [email protected] Min Zhang, King’s College London British Heart Foundation Centre of Research Excellence, School of Cardiovascular and Metabolic Medicine & Sciences, London, SE5 9NU, UK, Email [email protected]: Pulmonary fibrosis (PF) is a chronic and progressive lung disease characterized by excessive scarring of lung tissue, ultimately leading to impaired pulmonary function and poor survival outcomes. Currently, effective treatment options remain limited, with lung transplantation being the only definitive therapy. Wogonin, a bioactive flavonoid derived from the traditional Chinese medicinal herb Scutellaria baicalensis, has demonstrated potent anti-fibrotic effects in both in vitro and in vivo studies. However, its clinical application is hindered by poor tissue specificity, resulting in inadequate accumulation at fibrotic sites and low systemic bioavailability.Methods: We developed a lung-targeted, wogonin-loaded lipid nanoparticle system (LNP-Wog). The stability and biocompatibility of LNP-Wog were systematically evaluated, and its anti-fibrotic efficacy was assessed in murine PF models. Proteomic analysis was conducted to identify key components of the LNP-associated “protein corona” responsible for lung targeting. Additionally, biotin-affinity pulldown assays combined with Gene Ontology (GO) enrichment analysis were performed to elucidate the underlying anti-fibrotic mechanisms of wogonin.Results: LNP-Wog exhibited excellent stability, biocompatibility, and significant anti-fibrotic efficacy in murine PF models. Proteomic analysis revealed fibrinogen as a critical component of the LNP “protein corona”, facilitating lung endothelial targeting through integrin-mediated interaction. Mechanistically, wogonin was found to localize to the endoplasmic reticulum, where it promotes proteostasis by inhibiting protein synthesis via enhanced phosphorylation of eIF2α, a key event in the integrated stress response.Conclusion: These findings underscore the therapeutic potential of lung-targeted LNP-Wog nanoparticles as a promising strategy for the treatment of pulmonary fibrosis.Keywords: pulmonary fibrosis, wogonin, lipid nanoparticles, protein synthesis, eIF2&#x03B1