A hierarchical assembly strategy to engineer dextran-enveloped polyurethane nanopolyplexes for robust ovarian cancer gene therapy

A hierarchical assembly strategy is herein investigated to generate bio-responsive, dextran-enveloped, bioreducible polyurethane nanopolyplexes for nonviral gene therapy against ovarian tumor. Initially, a group of poly(urethane amine)s were designed and characterized for in vitro gene transfection. The polyurethane containing 1,4-bis(3-aminopropyl)piperazine residue (PUBAP) could induce the best in vitro transfection efficacy against SKOV-3 or A2780 ovarian cancer cells. Next, dextran-enveloped PUBAP polyplexes (e-polyplexes) were constructed by a hierarchical assembly procedure involving gene neutralization with PUBAP and subsequent gene condensation with a cationic dextran (SSDP800). Such dextran comprised dextran (15 kDa) as the main chain and multiple disulfide-linked branched polyethylenimine (BPEI) oligomers as the side grafts. Additionally, folate-dextran-enveloped PUBAP polyplexes (FA-e-polyplexes) were fabricated by folate-modified SSDP800. These nanoscale-enveloped polyplexes elicited an improved colloidal stability against salt ions and negatively charged heparin, efficient endosomal escaping, and bioreduction-triggered intracellular gene release. In vitro transfection against SKOV-3 cells illustrated that FA-e-polyplexes exerted higher transfection efficiency in the serum than e-polyplexes and 25 kDa BPEI-polyplexes. In vivo, FA-e-polyplexes yielded higher transgene expression level than e-polyplexes in an SKOV-3 tumor-bearing nude mouse model. In the tumor gene therapy with a small hairpin RNA silencing vascular endothelial growth factor, FA-e-polyplexes afforded higher tumor growth inhibition than polyplexes of folate-PEGylated PUBAP and 25 kDa linear polyethylenimine as positive controls. Importantly, such gene therapy had minor toxic effects on the health of the mouse. This work highlights a practical hierarchical assembly method to construct innovative enveloped polyurethane nanopolyplexes enabling robust ovarian cancer gene therapy. Statement of significance It is indispensable to rationally update binary cationic polyplexes into ternary polyplexes for vigorous tumor gene therapy. In this work, we have confirmed that a hierarchical assembly strategy, by using initial gene neutralization and subsequent gene condensation, is facile and effective to promote cationic polyurethane polyplexes into ternary folate-dextran-enveloped polyurethane polyplexes with a relatively high gene-loading capacity. The enveloped polyplex system enables more efficient gene transfection than the PEGylated polyplex counterpart in ovarian cancer in vitro and in vivo, thereby affording robust ovarian cancer gene therapy. The development of innovative enveloped polyplexes may be a new direction for a non-viral gene delivery system.