title: Melittin-grafted HPMA-oligolysine based copolymers for gene delivery.
abstract: Non-viral gene delivery systems capable of transfecting cells in the brain are critical in realizing the potential impact of nucleic acid therapeutics for diseases of the central nervous system. In this study, the membrane-lytic peptide melittin was incorporated into block copolymers synthesized by reversible addition-fragmentation chain transfer (RAFT) polymerization. The first block, designed for melittin conjugation, was composed of N-(2-hydroxypropyl)methacrylamide (HPMA) and pyridyl disulfide methacrylamide (PDSMA) and the second block, designed for DNA binding, was composed of oligo-l-lysine (K10) and HPMA. Melittin modified with cysteine at the C-terminus was conjugated to the polymers through the pyridyl disulfide pendent groups via disulfide exchange. The resulting pHgMelbHK10 copolymers are more membrane-lytic than melittin-free control polymers, and efficiently condensed plasmid DNA into salt-stable particles (~100-200 nm). The melittin-modified polymers transfected both HeLa and neuron-like PC-12 cells more efficiently than melittin-free polymers although toxicity associated with the melittin peptide was observed. Optimized formulations containing the luciferase reporter gene were delivered to mouse brain by intraventricular brain injections. Melittin-containing polyplexes produced about 35-fold higher luciferase activity in the brain compared to polyplexes without melittin. Thus, the melittin-containing block copolymers described in this work are promising materials for gene delivery to the brain.
authors.affiliation: Department of Bioengineering, Univeristy of Washington, Seattle, WA 98195, USA.
authors.forename: Joan G
authors.initials: JG
authors.lastname: Schellinger
authors.forename: Joshuel A
authors.initials: JA
authors.lastname: Pahang
authors.forename: Russell N
authors.initials: RN
authors.lastname: Johnson
authors.forename: David S H
authors.initials: DS
authors.lastname: Chu
authors.forename: Drew L
authors.initials: DL
authors.lastname: Sellers
authors.forename: Don O
authors.initials: DO
authors.lastname: Maris
authors.forename: Anthony J
authors.initials: AJ
authors.lastname: Convertine
authors.forename: Patrick S
authors.initials: PS
authors.lastname: Stayton
authors.forename: Philip J
authors.initials: PJ
authors.lastname: Horner
authors.forename: Suzie H
authors.initials: SH
authors.lastname: Pun
mesh_terms: D000178:Acrylamides / Q000737:chemistry; D000818:Animals; D001921:Brain / Q000187:drug effects / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000737:chemistry; D005260:Female; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D015316:Genetic Therapy / Q000379:methods*; D006367:HeLa Cells; D006801:Humans; D008156:Luciferases / Q000378:metabolism; D008239:Lysine / Q000032:analysis / Q000378:metabolism; D008555:Melitten / Q000737:chemistry*; D008689:Methacrylates / Q000737:chemistry*; D051379:Mice; D008810:Mice, Inbred C57BL; D046529:Microscopy, Electron, Transmission; D016716:PC12 Cells; D058105:Polymerization; D011108:Polymers / Q000737:chemistry*; D051381:Rats; D014162:Transfection
chemical_list: D000178:Acrylamides; D004268:DNA-Binding Proteins; D008689:Methacrylates; D011108:Polymers; D008555:Melitten; D008156:Luciferases; D008239:Lysine; C045985:methacrylamide; C032976:N-(2-hydroxypropyl)methacrylamide; C032802:hydroxypropyl methacrylate