Boosting RNA nanotherapeutics with V-ATPase activating non-inflammatory lipid nanoparticles to treat chronic lung injury
Zhiqiang Zhao, Xinzhu Shan, Jing Ding, Bin Ma, Buyao Li, Wendi Huang, Qingqing Yang, Yian Fang, J. C. Chen, Chenglin Song, Chenlong Wei, Shuai Liu, Xingdi Cheng, Shengran Zhang, Yunxuan Liu, Hongkun Wu, Cong Luo, Shaokun Shu, Xue Qiao, Zefeng Wang, Xueguang Lu, Lei Miao
Abstract
Lipid nanoparticles (LNPs) are a promising platform for mRNA delivery. However, their use in inflammatory pulmonary diseases is limited by reactogenicity and suboptimal delivery. Here we develop a non-inflammatory LNP (NIF-LNP) by incorporating ursolic acid, identified from a natural product library, into a biodegradable, cationic phosphoramide-derived LNP formulation. NIF-LNPs exhibit a 40-fold enhancement in lung protein expression without causing significant reactogenicity compared to LNPs containing ALC-0315. Our CRISPR-KO mechanistic studies uncover that ursolic acid promote endosome acidification by activating the V-ATPase complex, acting as a central hub for endosomal trafficking of LNPs and inflammation control. Furthermore, we identify an intracellular circadian regulatory gene, NR1D1, encapsulated in NIF-LNPs, showing notable therapeutic efficacy in bronchopulmonary dysplasia and lung fibrosis. To enhance clinical feasibility, we have developed a lyophilized formulation that maintains stability for over 90 days and ensures efficient nebulization in preclinical male mouse, pup rat, and male dog models. Overall, this V-ATPase-activating atomized NIF-LNP presents a viable strategy for treating variable chronic inflammatory lung diseases. Lipid nanoparticles (LNPs) are a promising platform for mRNA delivery, but their use in inflammatory pulmonary diseases is limited by reactogenicity and suboptimal delivery. Here, the authors report a VATPase activating non-inflammatory LNP (NIF-LNP) exhibiting a 40fold enhancement in lung protein expression without causing significant reactogenicity compared to commercial LNPs.