Systematic development of ionizable lipid nanoparticles for placental mRNA delivery using a design of experiments approach

Rachel E Young; Katherine M Nelson; Samuel I Hofbauer; Tara Vijayakumar; Mohamad-Gabriel Alameh; Drew Weissman; Charalampos Papachristou; Jason P Gleghorn; Rachel S Riley

doi:10.1016/j.bioactmat.2023.11.014

Systematic development of ionizable lipid nanoparticles for placental mRNA delivery using a design of experiments approach

Bioact Mater. 2023 Dec 22:34:125-137. doi: 10.1016/j.bioactmat.2023.11.014. eCollection 2024 Apr.

Authors

Rachel E Young^{1

2}, Katherine M Nelson³, Samuel I Hofbauer^{1

2

4}, Tara Vijayakumar^{1

2}, Mohamad-Gabriel Alameh⁵, Drew Weissman⁵, Charalampos Papachristou⁶, Jason P Gleghorn⁷, Rachel S Riley^{1

2}

Affiliations

¹ Department of Biomedical Engineering, Henry M. Rowan College of Engineering, Rowan University, 201 Mullica Hill Rd, Glassboro, NJ 08028, United States.
² School of Translational Biomedical Engineering & Sciences, Virtua College of Medicine & Life Sciences of Rowan University, 201 Mullica Hill Rd, Glassboro, NJ 08028, United States.
³ Department of Chemical and Biomolecular Engineering, College of Engineering, University of Delaware, 150 Academy Street, Newark, DE 19716, United States.
⁴ Cooper Medical School of Rowan University, Rowan University, 401 Broadway, Camden, NJ 08103, United States.
⁵ Perelman School of Medicine, University of Pennsylvania, 3400 Civic Center Blvd, Philadelphia, PA 19104, United States.
⁶ Department of Mathematics, College of Science & Mathematics, Rowan University, 201 Mullica Hill Rd, Glassboro, NJ 08028, United States.
⁷ Department of Biomedical Engineering, College of Engineering, University of Delaware, 590 Avenue 1743, Newark, DE 19713, United States.

Abstract

Ionizable lipid nanoparticles (LNPs) have gained attention as mRNA delivery platforms for vaccination against COVID-19 and for protein replacement therapies. LNPs enhance mRNA stability, circulation time, cellular uptake, and preferential delivery to specific tissues compared to mRNA with no carrier platform. However, LNPs are only in the beginning stages of development for safe and effective mRNA delivery to the placenta to treat placental dysfunction. Here, we develop LNPs that enable high levels of mRNA delivery to trophoblasts in vitro and to the placenta in vivo with no toxicity. We conducted a Design of Experiments to explore how LNP composition, including the type and molar ratio of each lipid component, drives trophoblast and placental delivery. Our data revealed that utilizing C12-200 as the ionizable lipid and 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) as the phospholipid in the LNP design yields high transfection efficiency in vitro. Analysis of lipid molar composition as a design parameter in LNPs displayed a strong correlation between apparent pKa and poly (ethylene) glycol (PEG) content, as a reduction in PEG molar amount increases apparent pKa. Further, we present one LNP platform that exhibits the highest delivery of placental growth factor mRNA to the placenta in pregnant mice, resulting in synthesis and secretion of a potentially therapeutic protein. Lastly, our high-performing LNPs have no toxicity to both the pregnant mice and fetuses. Our results demonstrate the feasibility of LNPs as a platform for mRNA delivery to the placenta, and our top LNP formulations may provide a therapeutic platform to treat diseases that originate from placental dysfunction during pregnancy.

Keywords: Drug delivery; Lipid nanoparticles (LNPs); Nucleic acids; Placenta; Placental growth factor (PlGF); Preeclampsia; Pregnancy.

Abstract

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