Engineering Lipid Nanoparticles for Enhanced Intracellular Delivery of mRNA through InhalationClick to copy article linkArticle link copied!
- Jeonghwan KimJeonghwan KimDepartment of Pharmaceutical Sciences, College of Pharmacy, Robertson Life Sciences Building, Oregon State University, Portland, Oregon 97201, United StatesMore by Jeonghwan Kim
- Antony JozicAntony JozicDepartment of Pharmaceutical Sciences, College of Pharmacy, Robertson Life Sciences Building, Oregon State University, Portland, Oregon 97201, United StatesMore by Antony Jozic
- Yuxin LinYuxin LinDepartment of Chemistry, Duke University, Durham, North Carolina 27708, United StatesMore by Yuxin Lin
- Yulia EygerisYulia EygerisDepartment of Pharmaceutical Sciences, College of Pharmacy, Robertson Life Sciences Building, Oregon State University, Portland, Oregon 97201, United StatesMore by Yulia Eygeris
- Elissa BloomElissa BloomDepartment of Pharmaceutical Sciences, College of Pharmacy, Robertson Life Sciences Building, Oregon State University, Portland, Oregon 97201, United StatesMore by Elissa Bloom
- Xiaochen TanXiaochen TanDepartment of Chemistry, Duke University, Durham, North Carolina 27708, United StatesMore by Xiaochen Tan
- Christopher AcostaChristopher AcostaDepartment of Pharmaceutical Sciences, College of Pharmacy, Robertson Life Sciences Building, Oregon State University, Portland, Oregon 97201, United StatesMore by Christopher Acosta
- Kelvin D. MacDonaldKelvin D. MacDonaldDepartment of Pediatrics, School of Medicine, Oregon Health and Science University, Portland, Oregon 97239, United StatesMore by Kelvin D. MacDonald
- Kevin D. WelsherKevin D. WelsherDepartment of Chemistry, Duke University, Durham, North Carolina 27708, United StatesMore by Kevin D. Welsher
- Gaurav Sahay*Gaurav Sahay*Email: [email protected]Department of Pharmaceutical Sciences, College of Pharmacy, Robertson Life Sciences Building, Oregon State University, Portland, Oregon 97201, United StatesDepartment of Biomedical Engineering, Robertson Life Sciences Building, Oregon Health Science University, Portland, Oregon 97239, United StatesDepartment of Ophthalmology, Casey Eye Institute, Oregon Health & Science University, Portland, Oregon 97239, United StatesMore by Gaurav Sahay
Abstract

Despite lipid nanoparticles’ (LNPs) success in the effective and safe delivery of mRNA vaccines, an inhalation-based mRNA therapy for lung diseases remains challenging. LNPs tend to disintegrate due to shear stress during aerosolization, leading to ineffective delivery. Therefore, LNPs need to remain stable through the process of nebulization and mucus penetration, yet labile enough for endosomal escape. To meet these opposing needs, we utilized PEG lipid to enhance the surficial stability of LNPs with the inclusion of a cholesterol analog, β-sitosterol, to improve endosomal escape. Increased PEG concentrations in LNPs enhanced the shear resistance and mucus penetration, while β-sitosterol provided LNPs with a polyhedral shape, facilitating endosomal escape. The optimized LNPs exhibited a uniform particle distribution, a polyhedral morphology, and a rapid mucosal diffusion with enhanced gene transfection. Inhaled LNPs led to localized protein production in the mouse lung without pulmonary or systemic toxicity. Repeated administration of these LNPs led to sustained protein production in the lungs. Lastly, mRNA encoding the cystic fibrosis transmembrane conductance regulator (CFTR) was delivered after nebulization to a CFTR-deficient animal model, resulting in the pulmonary expression of this therapeutic protein. This study demonstrated the rational design approach for clinical translation of inhalable LNP-based mRNA therapies.
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