Design of Ionizable Lipids To Overcome the Limiting Step of Endosomal Escape: Application in the Intracellular Delivery of mRNA, DNA, and siRNA.

The intracellular delivery of nucleic acid molecules is a complex process involving several distinct steps; among these the endosomal escape appeared to be of particular importance for an efficient protein production (or inhibition) into host cells. In the present study, a new series of ionizable vectors, derived from naturally occurring aminoglycoside tobramycin, was prepared using improved synthetic procedures that allow structural variations on the linker and hydrophobic domain levels. Complexes formed between the new ionizable lipids and mRNA, DNA, or siRNA were characterized by cryo-TEM experiments and their transfection potency was evaluated using different cell types. We demonstrated that lead molecule 30, bearing a biodegradable diester linker, formed small complexes with nucleic acids and provided very high transfection efficiency with all nucleic acids and cell types tested. The obtained results suggested that the improved and "universal" delivery properties of 30 resulted from an optimized endosomal escape, through the lipid-mixing mechanism.

[1]  J. Ruysschaert,et al.  Formation and intracellular trafficking of lipoplexes and polyplexes. , 2005, Molecular therapy : the journal of the American Society of Gene Therapy.

[2]  Daniel G. Anderson,et al.  Knocking down barriers: advances in siRNA delivery , 2009, Nature Reviews Drug Discovery.

[3]  M. Al‐Rubeai,et al.  Introduction to viral vectors. , 2011, Methods in molecular biology.

[4]  I. Toth,et al.  Endosome Escape Strategies for Improving the Efficacy of Oligonucleotide Delivery Systems. , 2015, Current medicinal chemistry.

[5]  J. Benoit,et al.  A review of the current status of siRNA nanomedicines in the treatment of cancer. , 2013, Biomaterials.

[6]  Gert Storm,et al.  Endosomal escape pathways for delivery of biologicals. , 2011, Journal of controlled release : official journal of the Controlled Release Society.

[7]  E. Canning,et al.  A triploblast origin for Myxozoa? , 1998, Nature.

[8]  Özlem Türeci,et al.  mRNA-based therapeutics — developing a new class of drugs , 2014, Nature Reviews Drug Discovery.

[9]  R. Mulligan,et al.  The basic science of gene therapy. , 1993, Science.

[10]  R. Leventis,et al.  Interactions of mammalian cells with lipid dispersions containing novel metabolizable cationic amphiphiles. , 1990, Biochimica et biophysica acta.

[11]  C. Mandl,et al.  RNA: the new revolution in nucleic acid vaccines. , 2013, Seminars in immunology.

[12]  David T. Curiel,et al.  Engineering targeted viral vectors for gene therapy , 2007, Nature Reviews Genetics.

[13]  Shubiao Zhang,et al.  Toxicity of cationic lipids and cationic polymers in gene delivery. , 2006, Journal of controlled release : official journal of the Controlled Release Society.

[14]  E. Junquera,et al.  Recent progress in gene therapy to deliver nucleic acids with multivalent cationic vectors. , 2016, Advances in colloid and interface science.

[15]  J. Lehn,et al.  Kanamycin A‐Derived Cationic Lipids as Vectors for Gene Transfection , 2005, Chembiochem : a European journal of chemical biology.

[16]  J. Behr,et al.  Gene transfer with synthetic cationic amphiphiles: prospects for gene therapy. , 1994, Bioconjugate chemistry.

[17]  Zheng-Rong Lu,et al.  Multifunctional cationic lipid-based nanoparticles facilitate endosomal escape and reduction-triggered cytosolic siRNA release. , 2014, Molecular pharmaceutics.

[18]  E. Junquera,et al.  Cationic lipids as transfecting agents of DNA in gene therapy. , 2014, Current topics in medicinal chemistry.

[19]  B. Pitard,et al.  Physicochemical parameters of non-viral vectors that govern transfection efficiency. , 2008, Current gene therapy.

[20]  P. Cullis,et al.  On the mechanism whereby cationic lipids promote intracellular delivery of polynucleic acids , 2001, Gene Therapy.

[21]  J. Lehn,et al.  Aminoglycoside-Derived Cationic Lipids for Gene Transfection: Synthesis of Kanamycin A Derivatives , 2003 .

[22]  J. Hughes,et al.  Introduction of a disulfide bond into a cationic lipid enhances transgene expression of plasmid DNA. , 1998, Biochemical and biophysical research communications.

[23]  Paul J Hergenrother,et al.  Targeting RNA with small molecules. , 2008, Chemical reviews.

[24]  G. F. Joyce,et al.  Direct observation of aminoglycoside-RNA interactions by surface plasmon resonance. , 1997, Journal of the American Chemical Society.

[25]  J. Lehn,et al.  Aminoglycoside‐derived cationic lipids as efficient vectors for gene transfection in vitro and in vivo , 2002, The journal of gene medicine.

[26]  J. Lehn,et al.  The design of cationic lipids for gene delivery. , 2005, Current pharmaceutical design.

[27]  Daniel G. Anderson,et al.  Non-viral vectors for gene-based therapy , 2014, Nature Reviews Genetics.

[28]  J. Lehn,et al.  Self-assembled lamellar complexes of siRNA with lipidic aminoglycoside derivatives promote efficient siRNA delivery and interference , 2007, Proceedings of the National Academy of Sciences.

[29]  M. Pistello,et al.  Viral vectors: a look back and ahead on gene transfer technology. , 2013, The new microbiologica.

[30]  M. Ilies,et al.  Synthetic nucleic acid delivery systems: present and perspectives. , 2015, Journal of medicinal chemistry.

[31]  Meredith A Mintzer,et al.  Nonviral vectors for gene delivery. , 2009, Chemical reviews.

[32]  Shubiao Zhang,et al.  The headgroup evolution of cationic lipids for gene delivery. , 2013, Bioconjugate chemistry.

[33]  Sarah Seifert,et al.  Image-based analysis of lipid nanoparticle–mediated siRNA delivery, intracellular trafficking and endosomal escape , 2013, Nature Biotechnology.

[34]  B. Pitard,et al.  Probing the in vitro mechanism of action of cationic lipid/DNA lipoplexes at a nanometric scale , 2010, Nucleic acids research.

[35]  I. Khalil,et al.  Uptake Pathways and Subsequent Intracellular Trafficking in Nonviral Gene Delivery , 2006, Pharmacological Reviews.

[36]  W. Walther,et al.  Viral Vectors for Gene Transfer , 2012, Drugs.

[37]  I. Zuhorn,et al.  Size-dependent internalization of particles via the pathways of clathrin- and caveolae-mediated endocytosis. , 2004, The Biochemical journal.

[38]  Leaf Huang,et al.  Recent advances in nonviral vectors for gene delivery. , 2012, Accounts of chemical research.

[39]  H Fessi,et al.  Gene therapy and DNA delivery systems. , 2014, International journal of pharmaceutics.

[40]  Nathan F. Bouxsein,et al.  Synthesis and characterization of degradable multivalent cationic lipids with disulfide-bond spacers for gene delivery. , 2011, Biochimica et biophysica acta.

[41]  F. Szoka,et al.  Mechanism of DNA release from cationic liposome/DNA complexes used in cell transfection. , 1996, Biochemistry.

[42]  Shubiao Zhang,et al.  Transfection efficiency of cationic lipids with different hydrophobic domains in gene delivery. , 2010, Bioconjugate chemistry.

[43]  J. Lehn,et al.  Paromomycin and neomycin B derived cationic lipids: synthesis and transfection studies. , 2012, Journal of controlled release : official journal of the Controlled Release Society.

[44]  D. Hoekstra,et al.  Cationic lipids, lipoplexes and intracellular delivery of genes. , 2006, Journal of controlled release : official journal of the Controlled Release Society.

[45]  C. Pichon,et al.  Gene transfer by chemical vectors, and endocytosis routes of polyplexes, lipoplexes and lipopolyplexes in a myoblast cell line. , 2012, Biomaterials.

[46]  Shubiao Zhang,et al.  Lipoplex morphologies and their influences on transfection efficiency in gene delivery. , 2007, Journal of controlled release : official journal of the Controlled Release Society.