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Diacetylenic lipids in the design of stable lipopolymers able to complex and protect plasmid DNA

Different viral and non-viral vectors have been designed to allow the delivery of nucleic acids in gene therapy. In general, non-viral vectors have been associated with increased safety for in vivo use; however, issues regarding their efficacy, toxicity and stability continue to drive further resear...

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Autores principales: Temprana, C. Facundo, Prieto, M. Jimena, Igartúa, Daniela E., Femia, A. Lis, Amor, M. Silvia, Alonso, Silvia del Valle
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5636127/
https://www.ncbi.nlm.nih.gov/pubmed/29020107
http://dx.doi.org/10.1371/journal.pone.0186194
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author Temprana, C. Facundo
Prieto, M. Jimena
Igartúa, Daniela E.
Femia, A. Lis
Amor, M. Silvia
Alonso, Silvia del Valle
author_facet Temprana, C. Facundo
Prieto, M. Jimena
Igartúa, Daniela E.
Femia, A. Lis
Amor, M. Silvia
Alonso, Silvia del Valle
author_sort Temprana, C. Facundo
collection PubMed
description Different viral and non-viral vectors have been designed to allow the delivery of nucleic acids in gene therapy. In general, non-viral vectors have been associated with increased safety for in vivo use; however, issues regarding their efficacy, toxicity and stability continue to drive further research. Thus, the aim of this study was to evaluate the potential use of the polymerizable diacetylenic lipid 1,2-bis(10,12-tricosadiynoyl)-sn-glycero-3-phosphocholine (DC(8,9)PC) as a strategy to formulate stable cationic lipopolymers in the delivery and protection of plasmid DNA. Cationic lipopolymers were prepared following two different methodologies by using DC(8,9)PC, 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), and the cationic lipids (CL) 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP), stearylamine (SA), and myristoylcholine chloride (MCL), in a molar ratio of 1:1:0.2 (DMPC:DC(8,9)PC:CL). The copolymerization methodology allowed obtaining cationic lipopolymers which were smaller in size than those obtained by the cationic addition methodology although both techniques presented high size stability over a 166-day incubation period at 4°C. Cationic lipopolymers containing DOTAP or MCL were more efficient in complexing DNA than those containing SA. Moreover, lipopolymers containing DOTAP were found to form highly stable complexes with DNA, able to resist serum DNAses degradation. Furthermore, neither of the cationic lipopolymers (with or without DNA) induced red blood cell hemolysis, although metabolic activity determined on the L-929 and Vero cell lines was found to be dependent on the cell line, the formulation and the presence of DNA. The high stability and DNA protection capacity as well as the reduced toxicity determined for the cationic lipopolymer containing DOTAP highlight the potential advantage of using lipopolymers when designing novel non-viral carrier systems for use in in vivo gene therapy. Thus, this work represents the first steps toward developing a cationic lipopolymer-based gene delivery system using polymerizable and cationic lipids.
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spelling pubmed-56361272017-10-30 Diacetylenic lipids in the design of stable lipopolymers able to complex and protect plasmid DNA Temprana, C. Facundo Prieto, M. Jimena Igartúa, Daniela E. Femia, A. Lis Amor, M. Silvia Alonso, Silvia del Valle PLoS One Research Article Different viral and non-viral vectors have been designed to allow the delivery of nucleic acids in gene therapy. In general, non-viral vectors have been associated with increased safety for in vivo use; however, issues regarding their efficacy, toxicity and stability continue to drive further research. Thus, the aim of this study was to evaluate the potential use of the polymerizable diacetylenic lipid 1,2-bis(10,12-tricosadiynoyl)-sn-glycero-3-phosphocholine (DC(8,9)PC) as a strategy to formulate stable cationic lipopolymers in the delivery and protection of plasmid DNA. Cationic lipopolymers were prepared following two different methodologies by using DC(8,9)PC, 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), and the cationic lipids (CL) 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP), stearylamine (SA), and myristoylcholine chloride (MCL), in a molar ratio of 1:1:0.2 (DMPC:DC(8,9)PC:CL). The copolymerization methodology allowed obtaining cationic lipopolymers which were smaller in size than those obtained by the cationic addition methodology although both techniques presented high size stability over a 166-day incubation period at 4°C. Cationic lipopolymers containing DOTAP or MCL were more efficient in complexing DNA than those containing SA. Moreover, lipopolymers containing DOTAP were found to form highly stable complexes with DNA, able to resist serum DNAses degradation. Furthermore, neither of the cationic lipopolymers (with or without DNA) induced red blood cell hemolysis, although metabolic activity determined on the L-929 and Vero cell lines was found to be dependent on the cell line, the formulation and the presence of DNA. The high stability and DNA protection capacity as well as the reduced toxicity determined for the cationic lipopolymer containing DOTAP highlight the potential advantage of using lipopolymers when designing novel non-viral carrier systems for use in in vivo gene therapy. Thus, this work represents the first steps toward developing a cationic lipopolymer-based gene delivery system using polymerizable and cationic lipids. Public Library of Science 2017-10-11 /pmc/articles/PMC5636127/ /pubmed/29020107 http://dx.doi.org/10.1371/journal.pone.0186194 Text en © 2017 Temprana et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Temprana, C. Facundo
Prieto, M. Jimena
Igartúa, Daniela E.
Femia, A. Lis
Amor, M. Silvia
Alonso, Silvia del Valle
Diacetylenic lipids in the design of stable lipopolymers able to complex and protect plasmid DNA
title Diacetylenic lipids in the design of stable lipopolymers able to complex and protect plasmid DNA
title_full Diacetylenic lipids in the design of stable lipopolymers able to complex and protect plasmid DNA
title_fullStr Diacetylenic lipids in the design of stable lipopolymers able to complex and protect plasmid DNA
title_full_unstemmed Diacetylenic lipids in the design of stable lipopolymers able to complex and protect plasmid DNA
title_short Diacetylenic lipids in the design of stable lipopolymers able to complex and protect plasmid DNA
title_sort diacetylenic lipids in the design of stable lipopolymers able to complex and protect plasmid dna
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5636127/
https://www.ncbi.nlm.nih.gov/pubmed/29020107
http://dx.doi.org/10.1371/journal.pone.0186194
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