title: Polyhistidine-PEG:DNA nanocomposites for gene delivery.
abstract: Complexation of plasmid DNA with polycations is a popular method by which to transfer therapeutic nucleic acid sequences to cells. One caveat of the approach is that the positive zeta potential of the complexes facilitates interaction with blood constituents, leading to serum protein adsorption and complement activation. As a countermeasure, investigators have developed polycations combined with polyethylene glycol (PEG) to create complexes with reduced protein adsorption potential. We have designed and synthesized PEG-polyhistidine conjugates to evaluate the material class as potential gene delivery vehicles. Two conjugate architectures (comb-shaped and linear A-B block copolymers) were synthesized and formulated with plasmid DNA. The complexes were characterized with respect to DNA complexation capacity, hydrodynamic diameter, zeta potential, in vitro cytotoxicity and transfection capacity in a model cell line. PEG content of the conjugate significantly influenced the hydrodynamic diameter of the DNA:conjugate composite in aqueous solution. For comb-shaped conjugates steric hindrance attributed to PEG led to a direct relationship between the PEG content and the complex size. Both architectures could condensed plasmid DNA into complexes with hydrodynamic diameters <150 nm. Complexation of DNA with the polyhistidine-PEG conjugates resulted in nanocomposites with negative zeta potentials that retarded DNase I-mediated hydrolysis, and all conjugates showed low cytotoxicity to macrophages cultured in vitro. The transfection efficiency was approximately equivalent to DNA:polylysine complexes. The formulation characteristics and low cytotoxicity suggest that polyhistidine-PEG conjugates may be useful for gene delivery.
authors.affiliation: School of Chemical and Biomolecular Engineering and the Biomedical Engineering Program, Cornell University, 270 Olin Hall, Ithaca, NY 14850, USA.
authors.forename: David
authors.initials: D
authors.lastname: Putnam
authors.forename: Alexander N
authors.initials: AN
authors.lastname: Zelikin
authors.forename: Vladimir A
authors.initials: VA
authors.lastname: Izumrudov
authors.forename: Robert
authors.initials: R
authors.lastname: Langer
mesh_terms: D000818:Animals; D001672:Biocompatible Materials / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D004247:DNA / Q000008:administration & dosage* / Q000235:genetics; D005821:Genetic Techniques; D006639:Histidine; D007700:Kinetics; D008264:Macrophages; D051379:Mice; D010957:Plasmids / Q000235:genetics*; D011092:Polyethylene Glycols
chemical_list: D001672:Biocompatible Materials; C033223:polyhistidine; D011092:Polyethylene Glycols; D006639:Histidine; D004247:DNA