title: Development and characterization of self-assembling nanoparticles using a bio-inspired amphipathic peptide for gene delivery.
abstract: The design of a non-viral gene delivery vehicle capable of delivering and releasing a functional nucleic acid cargo intracellularly remains a formidable challenge. For systemic gene therapy to be successful a delivery vehicle is required that protects the nucleic acid cargo from enzymatic degradation, extravasates from the vasculature, traverses the cell membrane, disrupts the endosomal vesicles and unloads the cargo at its destination site, namely the nucleus for the purposes of gene delivery. This manuscript reports the extensive investigation of a novel amphipathic peptide composed of repeating RALA units capable of overcoming the biological barriers to gene delivery both in vitro and in vivo. Our data demonstrates the spontaneous self-assembly of cationic DNA-loaded nanoparticles when the peptide is complexed with pDNA. Nanoparticles were <100nm, were stable in the presence of serum and were fusogenic in nature, with increased peptide α-helicity at a lower pH. Nanoparticles proved to be non-cytotoxic, readily traversed the plasma membrane of both cancer and fibroblast cell lines and elicited reporter-gene expression following intravenous delivery in vivo. The results of this study indicate that RALA presents an exciting delivery platform for the systemic delivery of nucleic acid therapeutics.
authors.affiliation: School of Pharmacy, Queen's University Belfast, 97 Lisburn Road, Belfast BT9 7BL, Northern Ireland, UK. Electronic address: h.mccarthy@qub.ac.uk.
authors.forename: Helen O
authors.initials: HO
authors.lastname: McCarthy
authors.affiliation: School of Pharmacy, Queen's University Belfast, 97 Lisburn Road, Belfast BT9 7BL, Northern Ireland, UK.
authors.forename: Joanne
authors.initials: J
authors.lastname: McCaffrey
authors.affiliation: School of Pharmacy, Queen's University Belfast, 97 Lisburn Road, Belfast BT9 7BL, Northern Ireland, UK.
authors.forename: Cian M
authors.initials: CM
authors.lastname: McCrudden
authors.affiliation: School of Pharmacy, Queen's University Belfast, 97 Lisburn Road, Belfast BT9 7BL, Northern Ireland, UK.
authors.forename: Aleksey
authors.initials: A
authors.lastname: Zholobenko
authors.affiliation: School of Pharmacy, Queen's University Belfast, 97 Lisburn Road, Belfast BT9 7BL, Northern Ireland, UK.
authors.forename: Ahlam A
authors.initials: AA
authors.lastname: Ali
authors.affiliation: School of Pharmacy, Queen's University Belfast, 97 Lisburn Road, Belfast BT9 7BL, Northern Ireland, UK.
authors.forename: John W
authors.initials: JW
authors.lastname: McBride
authors.affiliation: School of Pharmacy, Queen's University Belfast, 97 Lisburn Road, Belfast BT9 7BL, Northern Ireland, UK.
authors.forename: Ashley S
authors.initials: AS
authors.lastname: Massey
authors.affiliation: School of Pharmacy, Queen's University Belfast, 97 Lisburn Road, Belfast BT9 7BL, Northern Ireland, UK.
authors.forename: Sreekanth
authors.initials: S
authors.lastname: Pentlavalli
authors.affiliation: School of Pharmacy, Queen's University Belfast, 97 Lisburn Road, Belfast BT9 7BL, Northern Ireland, UK.
authors.forename: Kun-Hung
authors.initials: KH
authors.lastname: Chen
authors.affiliation: School of Pharmacy, Queen's University Belfast, 97 Lisburn Road, Belfast BT9 7BL, Northern Ireland, UK.
authors.forename: Grace
authors.initials: G
authors.lastname: Cole
authors.affiliation: School of Pharmacy, Queen's University Belfast, 97 Lisburn Road, Belfast BT9 7BL, Northern Ireland, UK.
authors.forename: Stephen P
authors.initials: SP
authors.lastname: Loughran
authors.affiliation: School of Mechanical & Aerospace Engineering, Queen's University Belfast, 123 Stranmillis Road, Belfast BT9 5AH, Northern Ireland, UK.
authors.forename: Nicholas J
authors.initials: NJ
authors.lastname: Dunne
authors.affiliation: School of Pharmacy, Queen's University Belfast, 97 Lisburn Road, Belfast BT9 7BL, Northern Ireland, UK.
authors.forename: Ryan F
authors.initials: RF
authors.lastname: Donnelly
authors.affiliation: School of Pharmacy, Queen's University Belfast, 97 Lisburn Road, Belfast BT9 7BL, Northern Ireland, UK.
authors.forename: Victoria L
authors.initials: VL
authors.lastname: Kett
authors.affiliation: School of Pharmacy, Queen's University Belfast, 97 Lisburn Road, Belfast BT9 7BL, Northern Ireland, UK.
authors.forename: Tracy
authors.initials: T
authors.lastname: Robson
mesh_terms: D000818:Animals; D002460:Cell Line; D045744:Cell Line, Tumor; D002942:Circular Dichroism; D004247:DNA / Q000008:administration & dosage* / Q000737:chemistry; D004912:Erythrocytes / Q000187:drug effects; D005260:Female; D018014:Gene Transfer Techniques; D006461:Hemolysis / Q000187:drug effects; D006801:Humans; D049409:Luciferases, Firefly / Q000235:genetics / Q000378:metabolism; D008810:Mice, Inbred C57BL; D053758:Nanoparticles / Q000008:administration & dosage* / Q000737:chemistry; D010316:Particle Size; D010455:Peptides / Q000008:administration & dosage* / Q000737:chemistry; D010957:Plasmids; D012756:Sheep
chemical_list: D010455:Peptides; C000596539:RALA peptide; D004247:DNA; D049409:Luciferases, Firefly
keywords: Amphipathic peptide; Biological barriers; Cell penetrating peptide; DNA delivery; Nanoparticle