Countless gene therapy vectors have been designed from viruses, lipids, and polymers to package, protect, and deliver genetic material into cells. A series of novel cationic polymers, (poly(2-methylacrylamido glucopyranose)-block-poly(2-methylaminoethylmethacrylate) (pMAG-b-pMAEMA), made in the Reineke group was designed to have superior biocompatibility and immune system stealth compared to other gene delivery vehicles. The purpose of this study was to quantify the biodistribution and hemocompatibility of these glycopolymers when complexed with plasmid DNA. Analysis via quantitative polymerase chain reaction (qPCR) showed widespread distribution of plasmid cargo in the spleen, heart, liver, lungs, kidneys, and brain following mouse tail intravenous injection. Fluorescently tagged polymers were also observed in the same organs, suggesting that the polyplexes are colloidally stable during circulation. An optical density hemolysis assay and visual observation determined that the polymer vectors did not lyse or affect red blood cell morphology up to 1.5 ng polymer per ul of blood. The prothrombin coagulation time (PT) was not affected by exposure to the complexes, but the activated partial thromboplastin coagulation time (aPTT) increased when plasma was incubated with complexes having high cationic polymer-to-DNA ratios. It was hypothesized that excess positive charge neutralizes the anionic aPTT reagent, causing coagulation to slow. The glycopolymer vectors demonstrated widespread distribution in vivo, suggesting systemic stability. The hemocompatibility of the complexes with lower polymer-DNA ratios was supported by the lack of significant red blood cell lysis or effects on coagulation. Future analysis of complement protein and platelet activation will offer a broad view on blood compatibility and increase the understanding of how polymeric gene delivery vehicles interact with biological components.

Hemocompatibility of colloidally stable glycopolymer-DNA complexes for gene therapy

249th National Meeting (2015)

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Hemocompatibility of colloidally stable glycopolymer-DNA complexes for gene therapy<br /> <br />