Date of Award
Spring 2023
Document Type
Dissertation
Degree Name
Doctor of Philosophy (PhD)
Department
Biomedical Engineering (ENAS)
First Advisor
Saltzman, Mark
Abstract
Advances in mRNA sequence design and chemical modifications have brought us to theprecipice of translating mRNA therapeutics to treat and cure innumerable human diseases. However, targeting mRNA delivery to relevant tissues remains a major obstacle in the clinical translation of mRNA therapeutics. Topical delivery of mRNA to the respiratory mucosa is an attractive strategy for treating pulmonary disorders. Despite significant interest, there are no approved delivery vehicles capable of efficiently and safely transfecting lung cells with mRNA. In this work, we created a polymer delivery vehicle for mRNA encapsulation and airway delivery. We employed a screening approach to formulate polyplexes with a library of biodegradable poly(amine-co-ester) (PACE) polymers. PACE-mRNA was shown to form small, stable polyplexes that efficiently encapsulate diverse RNA cargos, protect RNA from RNase degradation, enable mucosal penetration and cellular uptake, and achieve protein production in lung cells. Our optimized PACE-mRNA formulation achieved high lung cell transfection following intratracheal and intranasal administration to the respiratory tract. Protein expression was observed in airway epithelial cells and antigen presenting cells. We evaluated the therapeutic potential of PACE-mRNA delivery for two major applications in the lung; gene editing and mucosal vaccination. By co-encapsulating Cas9 mRNA and sgRNA, we demonstrated that PACE polyplexes achieve editing in the lung, primarily in small airway epithelial cells. We then encapsulated spike protein mRNA and demonstratedthat mucosal vaccination with PACE polyplexes induces lung resident and systemic cellular and humoral immunity against SARS-CoV-2. To our knowledge, this work represents the first reported mRNA-based vaccine to induce protective immunity against SARS-CoV-2 following mucosal administration.
Recommended Citation
Suberi, Alexandra Ahova, "Pulmonary Delivery of mRNA Therapeutics" (2023). Yale Graduate School of Arts and Sciences Dissertations. 1090.
https://elischolar.library.yale.edu/gsas_dissertations/1090
