正电改构铁蛋白纳米颗粒增强腺病毒载体细胞递送效率OA

Objective To develop positively charged human ferritin nanoparticles capable of modulating the surface properties of adenovirus vectors via electrostatic interaction-mediated self-assembly in order to enhance the cellular delivery efficiency of adenovirus vectors.Methods Based on structure-guided rational design,combinatorial amino acid mutations were introduced into wild-type human ferritin heavy chain(hF-wt)to generate a panel of positively charged ferritin mutants[(+)hFm].The particle size and electrostatic binding capacity of(+)hFm were characterized using dynamic light scattering(DLS)and agarose gel electrophoresis,respectively,to identify the optimal(+)hFm that could self-assemble into nanoparticles and exhibit a high affinity for negatively charged biomacromolecules.Furthermore,the properties of the complex formed by the adenovirus type 5 vector(Ad5)and(+)hFm[Ad5@(+)hFm]were determined.The efficacy with which target transgene expressions were enhanced and the potential cytotoxicity of Ad5@(+)hFm were subsequently evaluated in HeLa cells.Results The positively charged ferritins constructed by combinatorial amino acid mutations successfully self-assembled into nanoparticles with size distributions ranging from 10 to 70 nm.Among these mutants,(+)hFm-2 showed strong electrostatic binding to plasmid DNA at a migration retardation rate of up to 95.59%.At a molar ratio of 1∶1000[Ad5 to(+)hFm-2],the two components formed a complex with modulated surface properties,which significantly improved Ad5-mediated transgene expressions in the cellular model without inducing detectable cytotoxicity.Conclusion The positively charged engineered ferritin variant(+)hFm-2 can efficiently bind to Ad5 via electrostatic interaction,enhance Ad5-mediated transgene expressions,and possess high biocompatibility.