Lipid-like nanoparticles(LLNs)have been extensively explored for messenger RNA(mRNA)delivery in various biomedical applications.However,the long-term storage of these nanoparticles is still a challenge for their clini...Lipid-like nanoparticles(LLNs)have been extensively explored for messenger RNA(mRNA)delivery in various biomedical applications.However,the long-term storage of these nanoparticles is still a challenge for their clinical translation.In this study,we investigated a series of conditions for the long-term storage of LLNs with encapsulation of mRNA.We evaluated the stability of LLNs with different concentrations of cryoprotectants(sucrose,trehalose or mannitol)under the conditions of freezing or lyophilization processes.Through in vitro and in vivo mRNA delivery studies,we identified the optimal storage condition,and found that the addition with 5%(w/v)sucrose or trehalose to LLNs could remain their mRNA delivery efficiency for at least three months in the liquid nitrogen storage condition.展开更多
Activated fibroblasts are major mediators of pulmonary fibrosis.Fibroblasts are generally found in the connective tissue but upon activation can generate excess extracellular matrix(ECM)in the lung interstitial sectio...Activated fibroblasts are major mediators of pulmonary fibrosis.Fibroblasts are generally found in the connective tissue but upon activation can generate excess extracellular matrix(ECM)in the lung interstitial section.Therefore,fibroblasts are one of the most targeted cells for treating idiopathic pulmonary fibrosis(IPF).Here,we develop an anti-fibrotic platform that can modulate both the lysophosphatidic acid receptor 1(LPA_(1))and the inflammatory pathway through tumor necrosis factorα-induced protein 3(TNFAIP3,also known as A20)in fibroblasts.First,we synthesized a series of LPA_(1) antagonists,AM095 and AM966,derived amino lipids(LA lipids)which were formulated into LA-lipid nanoparticles(LA-LNPs)encapsulating mRNA.Specifically,LA5-LNPs,with AM966 head group and biodegradable acetal lipid tails,showed efficient A20 mRNA delivery to lung fibroblasts in vitro(80.2%±1.5%)and ex vivo(17.2%±0.4%).When treated to primary mouse lung fibroblasts(MLF),this formulation inhibited fibroblast migration and collagen production,thereby slowing the progression of IPF.Overall,LA5-LNPs encapsulated with A20 mRNA is a novel platform offering a potential approach to regulate fibroblast activation for the treatment of IPF.展开更多
基金supported by the Bayer Hemophilia Awards Program as well as the start-up fund from the College of Pharmacy at The Ohio State University.
文摘Lipid-like nanoparticles(LLNs)have been extensively explored for messenger RNA(mRNA)delivery in various biomedical applications.However,the long-term storage of these nanoparticles is still a challenge for their clinical translation.In this study,we investigated a series of conditions for the long-term storage of LLNs with encapsulation of mRNA.We evaluated the stability of LLNs with different concentrations of cryoprotectants(sucrose,trehalose or mannitol)under the conditions of freezing or lyophilization processes.Through in vitro and in vivo mRNA delivery studies,we identified the optimal storage condition,and found that the addition with 5%(w/v)sucrose or trehalose to LLNs could remain their mRNA delivery efficiency for at least three months in the liquid nitrogen storage condition.
基金the Maximizing Investigators’Research Award(No.R35GM119679)the National Institute of General Medical Sciences(No.R35GM144117)+1 种基金the support from the Professor Sylvan G.Frank Graduate Fellowshipthe Presidential Fellowship.
文摘Activated fibroblasts are major mediators of pulmonary fibrosis.Fibroblasts are generally found in the connective tissue but upon activation can generate excess extracellular matrix(ECM)in the lung interstitial section.Therefore,fibroblasts are one of the most targeted cells for treating idiopathic pulmonary fibrosis(IPF).Here,we develop an anti-fibrotic platform that can modulate both the lysophosphatidic acid receptor 1(LPA_(1))and the inflammatory pathway through tumor necrosis factorα-induced protein 3(TNFAIP3,also known as A20)in fibroblasts.First,we synthesized a series of LPA_(1) antagonists,AM095 and AM966,derived amino lipids(LA lipids)which were formulated into LA-lipid nanoparticles(LA-LNPs)encapsulating mRNA.Specifically,LA5-LNPs,with AM966 head group and biodegradable acetal lipid tails,showed efficient A20 mRNA delivery to lung fibroblasts in vitro(80.2%±1.5%)and ex vivo(17.2%±0.4%).When treated to primary mouse lung fibroblasts(MLF),this formulation inhibited fibroblast migration and collagen production,thereby slowing the progression of IPF.Overall,LA5-LNPs encapsulated with A20 mRNA is a novel platform offering a potential approach to regulate fibroblast activation for the treatment of IPF.
