Optimization of lipid nanoparticles for gene editing of the liver via intraduodenal delivery

Yining Zhu; Shuting Sarah Cai; Jingyao Ma; Leonardo Cheng; Christine Wei; Ataes Aggarwal; Wu Han Toh; Charles Shin; Ruochen Shen; Jiayuan Kong; Shuming Alan Mao; Yeh-Hsing Lao; Kam W Leong; Hai-Quan Mao

doi:10.1016/j.biomaterials.2024.122559

Optimization of lipid nanoparticles for gene editing of the liver via intraduodenal delivery

Biomaterials. 2024 Jul:308:122559. doi: 10.1016/j.biomaterials.2024.122559. Epub 2024 Apr 4.

Authors

Affiliations

¹ Department of Biomedical Engineering, Johns Hopkins University School of Medicine, Baltimore, MD, 21218, USA; Institute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD, 21218, USA; Translational Tissue Engineering Center, Johns Hopkins University School of Medicine, Baltimore, MD, 21218, USA.
² Department of Biomedical Engineering, Columbia University, New York, NY, 10027, USA.
³ Institute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD, 21218, USA; Translational Tissue Engineering Center, Johns Hopkins University School of Medicine, Baltimore, MD, 21218, USA; Department of Materials Science and Engineering, Johns Hopkins University, Baltimore, MD, 21218, USA.
⁴ Department of Biomedical Engineering, Johns Hopkins University School of Medicine, Baltimore, MD, 21218, USA; Institute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD, 21218, USA; Department of Computer Science, Johns Hopkins University, Baltimore, MD, 21218, USA.
⁵ Institute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD, 21218, USA; Department of Computer Science, Johns Hopkins University, Baltimore, MD, 21218, USA; Department of Biology, Johns Hopkins University, Baltimore, MD, 21218, USA.
⁶ Department of Biomedical Engineering, Johns Hopkins University School of Medicine, Baltimore, MD, 21218, USA; Institute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD, 21218, USA.
⁷ Department of Biomedical Engineering, Johns Hopkins University School of Medicine, Baltimore, MD, 21218, USA; Department of Computer Science, Johns Hopkins University, Baltimore, MD, 21218, USA.
⁸ Department of Biomedical Engineering, Columbia University, New York, NY, 10027, USA; Department of Systems Biology, Columbia University Medical Center, New York, NY, 10032, USA. Electronic address: kam.leong@columbia.edu.
⁹ Department of Biomedical Engineering, Johns Hopkins University School of Medicine, Baltimore, MD, 21218, USA; Institute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD, 21218, USA; Translational Tissue Engineering Center, Johns Hopkins University School of Medicine, Baltimore, MD, 21218, USA; Department of Materials Science and Engineering, Johns Hopkins University, Baltimore, MD, 21218, USA. Electronic address: hmao@jhu.edu.

PMID: 38583366
DOI: 10.1016/j.biomaterials.2024.122559

Abstract

Lipid nanoparticles (LNPs) have recently emerged as successful gene delivery platforms for a diverse array of disease treatments. Efforts to optimize their design for common administration methods such as intravenous injection, intramuscular injection, or inhalation, revolve primarily around the addition of targeting ligands or the choice of ionizable lipid. Here, we employed a multi-step screening method to optimize the type of helper lipid and component ratios in a plasmid DNA (pDNA) LNP library to efficiently deliver pDNA through intraduodenal delivery as an indicative route for oral administration. By addressing different physiological barriers in a stepwise manner, we down-selected effective LNP candidates from a library of over 1000 formulations. Beyond reporter protein expression, we assessed the efficiency in non-viral gene editing in mouse liver mediated by LNPs to knockdown PCSK9 and ANGPTL3 expression, thereby lowering low-density lipoprotein (LDL) cholesterol levels. Utilizing an all-in-one pDNA construct with Strep. pyogenes Cas9 and gRNAs, our results showcased that intraduodenal administration of selected LNPs facilitated targeted gene knockdown in the liver, resulting in a 27% reduction in the serum LDL cholesterol level. This LNP-based all-in-one pDNA-mediated gene editing strategy highlights its potential as an oral therapeutic approach for hypercholesterolemia, opening up new possibilities for DNA-based gene medicine applications.

Keywords: Hypercholesterolemia; Intraduodenal injection; Lipid nanoparticles; Liver gene editing; Oral delivery; Plasmid DNA.

Publication types

Research Support, Non-U.S. Gov't
Research Support, N.I.H., Extramural

MeSH terms

Animals
DNA / administration & dosage
DNA / genetics
Duodenum / metabolism
Gene Editing* / methods
Gene Transfer Techniques
Humans
Lipids* / chemistry
Liver* / metabolism
Mice
Mice, Inbred C57BL
Nanoparticles* / chemistry
Plasmids / administration & dosage
Plasmids / genetics
Proprotein Convertase 9 / genetics
Proprotein Convertase 9 / metabolism

Substances

Lipids
Proprotein Convertase 9
DNA