Inhaled Nanoformulated MRNA Polyplexes for Protein Production in Lung Epithelium

Overview

Journal Adv Mater

Date 2019 Jan 5

PMID 30609147

Citations 145

Authors

Asha Kumari Patel

James C Kaczmarek

Suman Bose

Kevin J Kauffman

Faryal Mir

Michael W Heartlein

Frank DeRosa

Robert Langer

Daniel G Anderson

Affiliations

Soon will be listed here.

Abstract

Noninvasive aerosol inhalation is an established method of drug delivery to the lung, and remains a desirable route for nucleic-acid-based therapeutics. In vitro transcribed (IVT) mRNA has broad therapeutic applicability as it permits temporal and dose-dependent control of encoded protein expression. Inhaled delivery of IVT-mRNA has not yet been demonstrated and requires development of safe and effective materials. To meet this need, hyperbranched poly(beta amino esters) (hPBAEs) are synthesized to enable nanoformulation of stable and concentrated polyplexes suitable for inhalation. This strategy achieves uniform distribution of luciferase mRNA throughout all five lobes of the lung and produces 101.2 ng g of luciferase protein 24 h after inhalation of hPBAE polyplexes. Importantly, delivery is localized to the lung, and no luminescence is observed in other tissues. Furthermore, using an Ai14 reporter mouse model it is identified that 24.6% of the total lung epithelial cell population is transfected after a single dose. Repeat dosing of inhaled hPBAE-mRNA generates consistent protein production in the lung, without local or systemic toxicity. The results indicate that nebulized delivery of IVT-mRNA facilitated by hPBAE vectors may provide a clinically relevant delivery system to lung epithelium.

Citing Articles

Biomineralize Mitochondria in Metal-Organic Frameworks to Promote Mitochondria Transplantation From Non-Tumorigenic Cells Into Cancer Cells.

Zhou J, Liu C, Wang Y, Guo Y, Xu X, Vuorimaa-Laukkanen E Smart Med. 2025; 4(1):e134.

PMID: 40059964 PMC: 11862567. DOI: 10.1002/smmd.134.

Advances and prospects of RNA delivery nanoplatforms for cancer therapy.

Attia M, Kijanka G, Nguyen N, Zhang J, An H Acta Pharm Sin B. 2025; 15(1):52-96.

PMID: 40041887 PMC: 11873661. DOI: 10.1016/j.apsb.2024.09.009.

Advanced drug delivery systems for the management of local conditions.

Lopez-Vidal L, Juskaite K, Ramoller I, Real D, McKenna P, Priotti J Ther Deliv. 2025; 16(3):285-303.

PMID: 40020739 PMC: 11875478. DOI: 10.1080/20415990.2024.2437978.

Highly branched poly β-amino ester/CpG-depleted plasmid nanoparticles for non-viral gene therapy in lung cystic fibrosis disease.

Qiu B, Manzanares D, Li Y, Wang X, Li Z, Terreau S Mol Ther Methods Clin Dev. 2025; 32(3):101292.

PMID: 40017666 PMC: 11866167. DOI: 10.1016/j.omtm.2024.101292.

Targeted delivery of TGF-β mRNA to murine lung parenchyma using one-component ionizable amphiphilic Janus Dendrimers.

Meshanni J, Stevenson E, Zhang D, Sun R, Ona N, Reagan E Nat Commun. 2025; 16(1):1806.

PMID: 39984450 PMC: 11845678. DOI: 10.1038/s41467-025-56448-y.

References

Eltoukhy A, Siegwart D, Alabi C, Rajan J, Langer R, Anderson D . Effect of molecular weight of amine end-modified poly(β-amino ester)s on gene delivery efficiency and toxicity. Biomaterials. 2012; 33(13):3594-603. PMC: 3326358. DOI: 10.1016/j.biomaterials.2012.01.046. View

Godbey W, Wu K, Mikos A . Size matters: molecular weight affects the efficiency of poly(ethylenimine) as a gene delivery vehicle. J Biomed Mater Res. 1999; 45(3):268-75. DOI: 10.1002/(sici)1097-4636(19990605)45:3<268::aid-jbm15>3.0.co;2-q. View

Cutlar L, Zhou D, Gao Y, Zhao T, Greiser U, Wang W . Highly Branched Poly(β-Amino Esters): Synthesis and Application in Gene Delivery. Biomacromolecules. 2015; 16(9):2609-17. DOI: 10.1021/acs.biomac.5b00966. View

Kowalski P, Capasso Palmiero U, Huang Y, Rudra A, Langer R, Anderson D . Ionizable Amino-Polyesters Synthesized via Ring Opening Polymerization of Tertiary Amino-Alcohols for Tissue Selective mRNA Delivery. Adv Mater. 2018; :e1801151. PMC: 6320729. DOI: 10.1002/adma.201801151. View

Zugates G, Peng W, Zumbuehl A, Jhunjhunwala S, Huang Y, Langer R . Rapid Optimization of Gene Delivery by Parallel End-modification of Poly(β-amino ester)s. Mol Ther. 2017; 15(7):1306-1312. DOI: 10.1038/sj.mt.6300132. View

Ernst N, Ulrichskotter S, Schmalix W, Radler J, Galneder R, Mayer E . Interaction of liposomal and polycationic transfection complexes with pulmonary surfactant. J Gene Med. 2000; 1(5):331-40. DOI: 10.1002/(SICI)1521-2254(199909/10)1:5<331::AID-JGM60>3.0.CO;2-8. View

Koshkina N, Agoulnik I, Melton S, Densmore C, Knight V . Biodistribution and pharmacokinetics of aerosol and intravenously administered DNA-polyethyleneimine complexes: optimization of pulmonary delivery and retention. Mol Ther. 2003; 8(2):249-54. DOI: 10.1016/s1525-0016(03)00177-1. View

Alton E, Armstrong D, Ashby D, Bayfield K, Bilton D, Bloomfield E . Repeated nebulisation of non-viral CFTR gene therapy in patients with cystic fibrosis: a randomised, double-blind, placebo-controlled, phase 2b trial. Lancet Respir Med. 2015; 3(9):684-691. PMC: 4673100. DOI: 10.1016/S2213-2600(15)00245-3. View

Beck S, Laube B, Barberena C, Fischer A, Adams R, Chesnut K . Deposition and expression of aerosolized rAAV vectors in the lungs of Rhesus macaques. Mol Ther. 2002; 6(4):546-54. DOI: 10.1006/mthe.2002.0698. View

10.

Densmore C, Orson F, Xu B, Kinsey B, Waldrep J, Hua P . Aerosol delivery of robust polyethyleneimine-DNA complexes for gene therapy and genetic immunization. Mol Ther. 2000; 1(2):180-8. DOI: 10.1006/mthe.1999.0021. View

11.

Potash A, Wallen T, Karp P, Ernst S, Moninger T, Gansemer N . Adenoviral gene transfer corrects the ion transport defect in the sinus epithelia of a porcine CF model. Mol Ther. 2013; 21(5):947-53. PMC: 3666638. DOI: 10.1038/mt.2013.49. View

12.

Park M, Kim H, Hwang C, Han K, Choi Y, Song S . Highly efficient gene transfer with degradable poly(ester amine) based on poly(ethylene glycol) diacrylate and polyethylenimine in vitro and in vivo. J Gene Med. 2007; 10(2):198-207. DOI: 10.1002/jgm.1139. View

13.

Tzeng S, Guerrero-Cazares H, Martinez E, Sunshine J, Quinones-Hinojosa A, Green J . Non-viral gene delivery nanoparticles based on poly(β-amino esters) for treatment of glioblastoma. Biomaterials. 2011; 32(23):5402-10. PMC: 3118545. DOI: 10.1016/j.biomaterials.2011.04.016. View

14.

Metzger R, Klein O, Martin G, Krasnow M . The branching programme of mouse lung development. Nature. 2008; 453(7196):745-50. PMC: 2892995. DOI: 10.1038/nature07005. View

15.

Johnson L, Olsen J, Sarkadi B, Moore K, Swanstrom R, Boucher R . Efficiency of gene transfer for restoration of normal airway epithelial function in cystic fibrosis. Nat Genet. 1992; 2(1):21-5. DOI: 10.1038/ng0992-21. View

16.

Rudolph C, Ortiz A, Schillinger U, Jauernig J, Plank C, Rosenecker J . Methodological optimization of polyethylenimine (PEI)-based gene delivery to the lungs of mice via aerosol application. J Gene Med. 2004; 7(1):59-66. DOI: 10.1002/jgm.646. View

17.

Johler S, Rejman J, Guan S, Rosenecker J . Nebulisation of IVT mRNA Complexes for Intrapulmonary Administration. PLoS One. 2015; 10(9):e0137504. PMC: 4564175. DOI: 10.1371/journal.pone.0137504. View

18.

Madisen L, Zwingman T, Sunkin S, Oh S, Zariwala H, Gu H . A robust and high-throughput Cre reporting and characterization system for the whole mouse brain. Nat Neurosci. 2009; 13(1):133-40. PMC: 2840225. DOI: 10.1038/nn.2467. View

19.

Griesenbach U, Pytel K, Alton E . Cystic Fibrosis Gene Therapy in the UK and Elsewhere. Hum Gene Ther. 2015; 26(5):266-75. PMC: 4442579. DOI: 10.1089/hum.2015.027. View

20.

Kauffman K, Oberli M, Dorkin J, Hurtado J, Kaczmarek J, Bhadani S . Rapid, Single-Cell Analysis and Discovery of Vectored mRNA Transfection In Vivo with a loxP-Flanked tdTomato Reporter Mouse. Mol Ther Nucleic Acids. 2018; 10:55-63. PMC: 5734870. DOI: 10.1016/j.omtn.2017.11.005. View