Optimization of in vivo DNA delivery with NickFect peptide vectors.
As the field of gene therapy progresses, an increasingly urgent need has arisen for efficient and non-toxic vectors for the in vivo delivery of nucleic acids. Cell-penetrating peptides (CPP) are very efficient transfection reagents in vitro, however, their application in vivo needs improvement. To enhance in vivo transfection we designed various CPPs based on previous knowledge of internalization studies and physiochemical properties of NickFect (NF) nanoparticles. We show that increment of the helicity of these Transportan10 analogues improves the transfection efficiency. We rationally design by modifying the net charge and the helicity of the CPP a novel amphipathic α-helical peptide NF55 for in vivo application. NF55 condenses DNA into stable nanoparticles that are resistant to protease degradation, promotes endosomal escape, and transfects the majority of cells in a large cell population. We demonstrate that NF55 mediates DNA delivery in vivo with gene induction efficiency that is comparable to commercial transfection reagents. In addition to gene induction in healthy mice, NF55/DNA nanoparticles showed promising tumor transfection in various mouse tumor models, including an intracranial glioblastoma model. The efficiency of NF55 to convey DNA specifically into tumor tissue increased even further after coupling a PEG2000 to the peptide via a disulphide-bond. Furthermore, a solid formulation of NF55/DNA displayed an excellent stability profile without additives or special storage conditions. Together, its high transfection efficacy and stability profile make NF55 an excellent vector for the delivery of DNA in vivo.
Laboratory of Molecular Biotechnology, Institute of Technology, University of Tartu, Nooruse, 50411 Tartu, Estonia. Electronic address: krista.freimann@ut.ee.
Krista
K
Freimann
Laboratory of Molecular Biotechnology, Institute of Technology, University of Tartu, Nooruse, 50411 Tartu, Estonia.
Piret
P
Arukuusk
Laboratory of Molecular Biotechnology, Institute of Technology, University of Tartu, Nooruse, 50411 Tartu, Estonia.
Kaido
K
Kurrikoff
Department of Neurochemistry, The Arrhenius Laboratories for Natural Sciences, Stockholm University, SE-10691 Stockholm, Sweden.
Luís Daniel Ferreira
LDF
Vasconcelos
Laboratory of Molecular Biotechnology, Institute of Technology, University of Tartu, Nooruse, 50411 Tartu, Estonia.
Kadi-Liis
KL
Veiman
Laboratory of Molecular Biotechnology, Institute of Technology, University of Tartu, Nooruse, 50411 Tartu, Estonia.
Julia
J
Uusna
Department of Developmental Biology, Institute of Molecular and Cell Biology, University of Tartu, Riia 23, 51010 Tartu, Estonia.
Helerin
H
Margus
Institute of Chemistry, University of Tartu, Ravila 14a, 50411 Tartu, Estonia.
Alfonso T
AT
Garcia-Sosa
Department of Developmental Biology, Institute of Molecular and Cell Biology, University of Tartu, Riia 23, 51010 Tartu, Estonia.
Margus
M
Pooga
Laboratory of Molecular Biotechnology, Institute of Technology, University of Tartu, Nooruse, 50411 Tartu, Estonia; Department of Neurochemistry, The Arrhenius Laboratories for Natural Sciences, Stockholm University, SE-10691 Stockholm, Sweden.
Ülo
Ü
Langel
D000595:Amino Acid Sequence; D002470:Cell Survival / Q000187:drug effects; D057846:Cell-Penetrating Peptides / Q000737:chemistry*; D004247:DNA / Q000008:administration & dosage* / Q000235:genetics; D004337:Drug Carriers / Q000737:chemistry*; D018014:Gene Transfer Techniques; D006367:HeLa Cells; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D053758:Nanoparticles / Q000737:chemistry*; D010957:Plasmids; D011487:Protein Conformation; D014162:Transfection
D057846:Cell-Penetrating Peptides; D004337:Drug Carriers; D004247:DNA
Cell penetrating peptide; In vivo gene delivery; Nanoparticles; PEGylation; Transfection; Tumor