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

C. FACUNDO TEMPRANA; M. JIMENA PRIETO; DANIELA E. IGARTUA; LIS FEMIA; SILVIA AMOR; SILVIA DEL VALLE ALONSO

PLOS ONE

PUBLIC LIBRARY SCIENCE

Lugar: San Francisco; Año: 2017

1932-6203

Different viral and non-viral vectors have been designed to allow the delivery of nucleicacids in gene therapy. In general, non-viral vectors have been associated with increasedsafety for in vivo use; however, issues regarding their efficacy, toxicity and stability continueto drive further research. Thus, the aim of this study was to evaluate the potential use of thepolymerizable diacetylenic lipid 1,2-bis(10,12-tricosadiynoyl)-sn-glycero-3-phosphocholine(DC8,9PC) 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 DC8,9PC, 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), and thecationic 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:DC8,9PC:CL).The copolymerization methodology allowed obtaining cationic lipopolymers which weresmaller 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 thosecontaining SA. Moreover, lipopolymers containing DOTAP were found to form highly stablecomplexes with DNA, able to resist serum DNAses degradation. Furthermore, neither of thecationic 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 onthe cell line, the formulation and the presence of DNA. The high stability and DNA protectioncapacity as well as the reduced toxicity determined for the cationic lipopolymer containingDOTAP highlight the potential advantage of using lipopolymers when designing novel nonviral carrier systems for use in in vivo gene therapy. Thus, this work represents the firststeps toward developing a cationic lipopolymer-based gene delivery system using polymerizable and cationic lipids.