title: Intracellular plasmid DNA delivery by self-assembled nanoparticles of amphiphilic PHML-b-PLLA-b-PHML copolymers and the endocytosis pathway analysis.
abstract: This work presents a new series of polycationic nanoparticles of (l-)-lysine conjugated amphiphilic triblock copolymer poly(hydroxyletheyl methacrylate-L-lysine)-b-poly(L-lactide)-b-poly(hydroxyletheyl methacrylate-L-lysine)s (PHML-b-PLLA-b-PHML) as potent low cytotoxic vectors for intracellular plasmid DNA delivery. First, the triblock PHML-b-PLLA-b-PHML copolymers were prepared via a combination of metal-free controlled ring opening polymerization and successive atom transfer radical polymerization. Then the cationic PHML-b-PLLA-b-PHML nanoparticles were further prepared by solution self-assembly. The particle size, zeta potential and morphology of as-prepared PHML-b-PLLA-b-PHML nanoparticles were characterized by dynamic light scattering and atomic force microscopy, respectively. The plasmid DNA binding affinities and polyplex stabilities were separately explored by agarose gel retardation and DNase I degradation assays. Then in vitro cytotoxicity and gene transfection efficiency of the PHML-b-PLLA-b-PHML nanoparticles vectors as well as relevant polyplex endocytosis pathway were investigated with H1299 cells. It was revealed that the PHML-b-PLLA-b-PHML nanoparticles exhibited low cytotoxicity, strong plasmid DNA binding affinity, high polyplex stability and efficient plasmid DNA transfection even under serum conditions (10% FBS). Moreover, the endocytosis analysis results disclosed that the PHML30-b-PLLA-b-PHML30nanoparticle/plasmid DNA polyplexes were predominantly involved in lipid-raft-mediated endocytosis pathway, similar to that of SV40 virus-based vectors.
authors.affiliation: Laboratory of Synthetic and Self-assembly Chemistry for Organic Functional Molecules, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Shanghai, China.
authors.forename: Jingjing
authors.initials: J
authors.lastname: Sun
authors.affiliation: Laboratory of Synthetic and Self-assembly Chemistry for Organic Functional Molecules, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Shanghai, China.
authors.forename: Ting
authors.initials: T
authors.lastname: Luo
authors.affiliation: Laboratory of Synthetic and Self-assembly Chemistry for Organic Functional Molecules, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Shanghai, China Department of Chemistry, Université de Montréal, Succursale Centre-ville, Montréal, Quebéc, Canada rlsheng@mail.sioc.ac.cn.
authors.forename: Ruilong
authors.initials: R
authors.lastname: Sheng
authors.affiliation: Laboratory of Synthetic and Self-assembly Chemistry for Organic Functional Molecules, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Shanghai, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Li
authors.affiliation: Laboratory of Synthetic and Self-assembly Chemistry for Organic Functional Molecules, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Shanghai, China.
authors.forename: Zhao
authors.initials: Z
authors.lastname: Wang
authors.affiliation: Laboratory of Synthetic and Self-assembly Chemistry for Organic Functional Molecules, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Shanghai, China.
authors.forename: Amin
authors.initials: A
authors.lastname: Cao
mesh_terms: D065966:Absorption, Physicochemical; D045744:Cell Line, Tumor; D003460:Crystallization / Q000379:methods; D004058:Diffusion; D004705:Endocytosis; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D021962:Membrane Microdomains / Q000737:chemistry / Q000378:metabolism; D053858:Metabolic Networks and Pathways; D008768:Methylmethacrylates / Q000737:chemistry*; D053769:Nanocapsules / Q000737:chemistry* / Q000648:ultrastructure; D009374:Neoplasms, Experimental / Q000737:chemistry / Q000235:genetics* / Q000473:pathology; D010316:Particle Size; D010957:Plasmids / Q000008:administration & dosage / Q000737:chemistry / Q000235:genetics*; D011091:Polyesters / Q000737:chemistry*; D014162:Transfection / Q000379:methods*
chemical_list: D008768:Methylmethacrylates; D053769:Nanocapsules; D011091:Polyesters; C021302:hydroxymethylmethacrylate; C033616:poly(lactide)
keywords: L-lysine; Poly(L-lactide); endocytosis pathway; gene delivery; serum-compatible