Piperazine-derived lipid nanoparticles deliver mRNA to immune cells in vivo
In humans, lipid nanoparticles (LNPs) have safely delivered therapeutic RNA to hepatocytes after systemic administration and to antigen-presenting cells after intramuscular injection. However, systemic RNA delivery to non-hepatocytes remains challenging, especially without targeting ligands such as...
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Published in | Nature communications Vol. 13; no. 1; p. 4766 |
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Main Authors | , , , , , , , , , , |
Format | Journal Article |
Language | English |
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Nature Publishing Group UK
15.08.2022
Nature Publishing Group Nature Portfolio |
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Abstract | In humans, lipid nanoparticles (LNPs) have safely delivered therapeutic RNA to hepatocytes after systemic administration and to antigen-presenting cells after intramuscular injection. However, systemic RNA delivery to non-hepatocytes remains challenging, especially without targeting ligands such as antibodies, peptides, or aptamers. Here we report that piperazine-containing ionizable lipids (Pi-Lipids) preferentially deliver mRNA to immune cells in vivo without targeting ligands. After synthesizing and characterizing Pi-Lipids, we use high-throughput DNA barcoding to quantify how 65 chemically distinct LNPs functionally delivered mRNA (i.e., mRNA translated into functional, gene-editing protein) in 14 cell types directly in vivo. By analyzing the relationships between lipid structure and cellular targeting, we identify lipid traits that increase delivery in vivo. In addition, we characterize Pi-A10, an LNP that preferentially delivers mRNA to the liver and splenic immune cells at the clinically relevant dose of 0.3 mg/kg. These data demonstrate that high-throughput in vivo studies can identify nanoparticles with natural non-hepatocyte tropism and support the hypothesis that lipids with bioactive small-molecule motifs can deliver mRNA in vivo.
Next-generation lipid nanoparticles that target non-hepatocytes could be important clinical tools. Using in vivo DNA barcoding, the authors identify piperazine-containing lipids deliver mRNA to immune cells without targeting ligands. |
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AbstractList | In humans, lipid nanoparticles (LNPs) have safely delivered therapeutic RNA to hepatocytes after systemic administration and to antigen-presenting cells after intramuscular injection. However, systemic RNA delivery to non-hepatocytes remains challenging, especially without targeting ligands such as antibodies, peptides, or aptamers. Here we report that piperazine-containing ionizable lipids (Pi-Lipids) preferentially deliver mRNA to immune cells in vivo without targeting ligands. After synthesizing and characterizing Pi-Lipids, we use high-throughput DNA barcoding to quantify how 65 chemically distinct LNPs functionally delivered mRNA (i.e., mRNA translated into functional, gene-editing protein) in 14 cell types directly in vivo. By analyzing the relationships between lipid structure and cellular targeting, we identify lipid traits that increase delivery in vivo. In addition, we characterize Pi-A10, an LNP that preferentially delivers mRNA to the liver and splenic immune cells at the clinically relevant dose of 0.3 mg/kg. These data demonstrate that high-throughput in vivo studies can identify nanoparticles with natural non-hepatocyte tropism and support the hypothesis that lipids with bioactive small-molecule motifs can deliver mRNA in vivo.
Next-generation lipid nanoparticles that target non-hepatocytes could be important clinical tools. Using in vivo DNA barcoding, the authors identify piperazine-containing lipids deliver mRNA to immune cells without targeting ligands. Next-generation lipid nanoparticles that target non-hepatocytes could be important clinical tools. Using in vivo DNA barcoding, the authors identify piperazine-containing lipids deliver mRNA to immune cells without targeting ligands. In humans, lipid nanoparticles (LNPs) have safely delivered therapeutic RNA to hepatocytes after systemic administration and to antigen-presenting cells after intramuscular injection. However, systemic RNA delivery to non-hepatocytes remains challenging, especially without targeting ligands such as antibodies, peptides, or aptamers. Here we report that piperazine-containing ionizable lipids (Pi-Lipids) preferentially deliver mRNA to immune cells in vivo without targeting ligands. After synthesizing and characterizing Pi-Lipids, we use high-throughput DNA barcoding to quantify how 65 chemically distinct LNPs functionally delivered mRNA (i.e., mRNA translated into functional, gene-editing protein) in 14 cell types directly in vivo. By analyzing the relationships between lipid structure and cellular targeting, we identify lipid traits that increase delivery in vivo. In addition, we characterize Pi-A10, an LNP that preferentially delivers mRNA to the liver and splenic immune cells at the clinically relevant dose of 0.3 mg/kg. These data demonstrate that high-throughput in vivo studies can identify nanoparticles with natural non-hepatocyte tropism and support the hypothesis that lipids with bioactive small-molecule motifs can deliver mRNA in vivo.Next-generation lipid nanoparticles that target non-hepatocytes could be important clinical tools. Using in vivo DNA barcoding, the authors identify piperazine-containing lipids deliver mRNA to immune cells without targeting ligands. In humans, lipid nanoparticles (LNPs) have safely delivered therapeutic RNA to hepatocytes after systemic administration and to antigen-presenting cells after intramuscular injection. However, systemic RNA delivery to non-hepatocytes remains challenging, especially without targeting ligands such as antibodies, peptides, or aptamers. Here we report that piperazine-containing ionizable lipids (Pi-Lipids) preferentially deliver mRNA to immune cells in vivo without targeting ligands. After synthesizing and characterizing Pi-Lipids, we use high-throughput DNA barcoding to quantify how 65 chemically distinct LNPs functionally delivered mRNA (i.e., mRNA translated into functional, gene-editing protein) in 14 cell types directly in vivo. By analyzing the relationships between lipid structure and cellular targeting, we identify lipid traits that increase delivery in vivo. In addition, we characterize Pi-A10, an LNP that preferentially delivers mRNA to the liver and splenic immune cells at the clinically relevant dose of 0.3 mg/kg. These data demonstrate that high-throughput in vivo studies can identify nanoparticles with natural non-hepatocyte tropism and support the hypothesis that lipids with bioactive small-molecule motifs can deliver mRNA in vivo. |
ArticleNumber | 4766 |
Author | Loughrey, David Hatit, Marine Z. C. Cid, Ada Del Ni, Huanzhen Santangelo, Philip J. Dahlman, James E. Lokugamage, Melissa P. Peck, Hannah E. Muralidharan, Abinaya Zhao, Kun Kim, YongTae |
Author_xml | – sequence: 1 givenname: Huanzhen surname: Ni fullname: Ni, Huanzhen organization: Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology – sequence: 2 givenname: Marine Z. C. orcidid: 0000-0002-5713-6596 surname: Hatit fullname: Hatit, Marine Z. C. organization: Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology – sequence: 3 givenname: Kun surname: Zhao fullname: Zhao, Kun organization: Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology, School of Pharmaceutical Sciences, Shandong University – sequence: 4 givenname: David surname: Loughrey fullname: Loughrey, David organization: Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology – sequence: 5 givenname: Melissa P. surname: Lokugamage fullname: Lokugamage, Melissa P. organization: Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology – sequence: 6 givenname: Hannah E. surname: Peck fullname: Peck, Hannah E. organization: Parker H. Petit Institute for Bioengineering and Bioscience, Georgia Institute of Technology – sequence: 7 givenname: Ada Del surname: Cid fullname: Cid, Ada Del organization: Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology – sequence: 8 givenname: Abinaya surname: Muralidharan fullname: Muralidharan, Abinaya organization: George W. Woodruff School of Mechanical Engineering, Georgia Institute of Technology, Institute for Electronics and Nanotechnology, Georgia Institute of Technology – sequence: 9 givenname: YongTae orcidid: 0000-0002-8835-8247 surname: Kim fullname: Kim, YongTae organization: Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology, Parker H. Petit Institute for Bioengineering and Bioscience, Georgia Institute of Technology, George W. Woodruff School of Mechanical Engineering, Georgia Institute of Technology, Institute for Electronics and Nanotechnology, Georgia Institute of Technology – sequence: 10 givenname: Philip J. orcidid: 0000-0001-7352-0339 surname: Santangelo fullname: Santangelo, Philip J. organization: Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology – sequence: 11 givenname: James E. orcidid: 0000-0001-7580-436X surname: Dahlman fullname: Dahlman, James E. email: james.dahlman@bme.gatech.edu organization: Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology |
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Snippet | In humans, lipid nanoparticles (LNPs) have safely delivered therapeutic RNA to hepatocytes after systemic administration and to antigen-presenting cells after... Next-generation lipid nanoparticles that target non-hepatocytes could be important clinical tools. Using in vivo DNA barcoding, the authors identify... |
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SubjectTerms | 42 42/109 42/62 49 631/45/147 631/61/2300 631/61/350/354 631/61/391 639/925/926/1051 Antibodies Antigen-presenting cells Antigens Aptamers Cellular structure Deoxyribonucleic acid DNA Gene sequencing Genetic modification Genome editing Hepatocytes Humanities and Social Sciences Immune system In vivo methods and tests Ligands Lipid structure Lipids mRNA multidisciplinary Nanoparticles Peptides Piperazine Ribonucleic acid RNA Science Science (multidisciplinary) Tropism |
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Title | Piperazine-derived lipid nanoparticles deliver mRNA to immune cells in vivo |
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