Multi-step screening of DNA/lipid nanoparticles and co-delivery with siRNA to enhance and prolong gene expression

Yining Zhu; Ruochen Shen; Ivan Vuong; Rebekah A Reynolds; Melanie J Shears; Zhi-Cheng Yao; Yizong Hu; Won June Cho; Jiayuan Kong; Sashank K Reddy; Sean C Murphy; Hai-Quan Mao

doi:10.1038/s41467-022-31993-y

Multi-step screening of DNA/lipid nanoparticles and co-delivery with siRNA to enhance and prolong gene expression

Nat Commun. 2022 Jul 25;13(1):4282. doi: 10.1038/s41467-022-31993-y.

Authors

Yining Zhu^{1

2

3}, Ruochen Shen^{1

2

3}, Ivan Vuong^{1

2

3}, Rebekah A Reynolds^{4

5}, Melanie J Shears^{4

5}, Zhi-Cheng Yao^{2

3

6}, Yizong Hu^{1

2

3}, Won June Cho^{2

7}, Jiayuan Kong^{1

2

3}, Sashank K Reddy^{1

2

8}, Sean C Murphy^{9

10

11

12}, Hai-Quan Mao^{13

14

15

16}

Affiliations

¹ Department of Biomedical Engineering, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
² Institute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD, USA.
³ Translational Tissue Engineering Center, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
⁴ Department of Laboratory Medicine and Pathology, University of Washington, Seattle, WA, USA.
⁵ Center for Emerging and Re-emerging Infectious Diseases, University of Washington, Seattle, WA, USA.
⁶ Department of Materials Science and Engineering, Johns Hopkins University, Baltimore, MD, USA.
⁷ Department of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, MD, USA.
⁸ Department of Plastic and Reconstructive Surgery, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
⁹ Department of Laboratory Medicine and Pathology, University of Washington, Seattle, WA, USA. murphysc@uw.edu.
¹⁰ Center for Emerging and Re-emerging Infectious Diseases, University of Washington, Seattle, WA, USA. murphysc@uw.edu.
¹¹ Department of Microbiology, University of Washington, Seattle, WA, USA. murphysc@uw.edu.
¹² Seattle Malaria Clinical Trials Center, Fred Hutch Cancer Research Center, Seattle, WA, USA. murphysc@uw.edu.
¹³ Department of Biomedical Engineering, Johns Hopkins University School of Medicine, Baltimore, MD, USA. hmao@jhu.edu.
¹⁴ Institute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD, USA. hmao@jhu.edu.
¹⁵ Translational Tissue Engineering Center, Johns Hopkins University School of Medicine, Baltimore, MD, USA. hmao@jhu.edu.
¹⁶ Department of Materials Science and Engineering, Johns Hopkins University, Baltimore, MD, USA. hmao@jhu.edu.

Abstract

Lipid nanoparticles hold great potential as an effective non-viral vector for nucleic acid-based gene therapy. Plasmid DNA delivery can result in extended transgene expression compared to mRNA-based technologies, yet there is a lack of systematic investigation into lipid nanoparticle compositions for plasmid DNA delivery. Here, we report a multi-step screening platform to identify optimized plasmid DNA lipid nanoparticles for liver-targeted transgene expression. To achieve this, we analyze the role of different helper lipids and component ratios in plasmid DNA lipid nanoparticle-mediated gene delivery in vitro and in vivo. Compared to mRNA LNPs and in vivo-jetPEI/DNA nanoparticles, the identified plasmid DNA lipid nanoparticles successfully deliver transgenes and mediate prolonged expression in the liver following intravenous administration in mice. By addressing different physiological barriers in a stepwise manner, this screening platform can efficiently down select effective lipid nanoparticle candidates from a lipid nanoparticle library of over 1000 formulations. In addition, we substantially extend the duration of plasmid DNA nanoparticle-mediated transgene expression using a DNA/siRNA co-delivery approach that targets transcription factors regulating inflammatory response pathways. This lipid nanoparticle-based co-delivery strategy further highlights the unique advantages of an extended transgene expression profile using plasmid DNA delivery and offers new opportunities for DNA-based gene medicine applications.

Publication types

Research Support, N.I.H., Extramural

MeSH terms

Animals
DNA / genetics
Gene Expression
Lipids*
Liposomes
Mice
Nanoparticles*
RNA, Messenger
RNA, Small Interfering / genetics

Substances

Lipid Nanoparticles
Lipids
Liposomes
RNA, Messenger
RNA, Small Interfering
DNA

Grants and funding

U01 AI155313/AI/NIAID NIH HHS/United States