NANOSCALE: Pharmaceutically Engineered Nanoparticles for the Targeted Delivery of Plasmid DNA

研究人员提出了一种新的方法，以药物工程的大小范围为5到50纳米的固体纳米颗粒含有非病毒基因治疗应用的质粒DNA。该方法潜在地克服了目前非病毒基因递送技术的局限性，即使用带正电荷的脂类或聚合物与带负电荷的质粒DNA在100至1000 nm的尺寸范围内形成更大的、不稳定的络合物。此外，使用微乳液前驱体方法来设计固体纳米颗粒，可以自发和可重复地制造出定义良好且均匀的固体纳米颗粒(5到50 nm)，而无需使用昂贵的或潜在的破坏性技术，包括高扭矩机械混合、微流态化、均质或研磨。提出的方法包括自发形成新型微乳液，其中分散的液滴直径通常为5-50 nm，是形成固体和稳定纳米颗粒的前体。除了质粒DNA之外，工程纳米颗粒系统还可以包含许多不同的材料，用于各种医疗和工程应用，如纳米磁铁和纳米传感器。该项目的具体目标是证明可以从新型微乳液前驱体中工程出含有质粒DNA的靶向固体纳米颗粒。该项目有三个具体目标，1)证明可以将质粒DNA掺入到稳定的微乳液中，并且可以从微乳液前体制备稳定的包含质粒DNA的固体纳米颗粒(5-50 nm)；2)表征固体纳米颗粒(即尺寸、表面电荷和孔隙率、DNA释放和稳定性)并证明固体纳米颗粒在生物流体中稳定并且可以在体外表达转基因基因；3)在固体纳米颗粒的表面引入细胞特异性配体并证明目标纳米颗粒的表面识别。

The investigators propose a novel method to pharmaceutically engineer solid nanoparticles in the size range of 5 to 50 nm containing plasmid DNA fornon-viral gene therapy applications. The method potentially overcomeslimitations of current non-viral gene delivery technology which uses positively-charged lipids or polymers to form larger, unstable complexeswith negatively-charged plasmid DNA in the size range of 100 to 1000 nm. Further, using the microemulsion precursor method to engineer solid nanoparticles, well-defined and uniform solid nanoparticles (5 to 50 nm)may be spontaneously and reproducibly made without the use of expensive or potentially damaging techniques involving high-torque mechanical mixing, microfluidization, homogenization, or milling. The proposed method involves the spontaneous formation of novel microemulsions wherein the dispersed droplets, which are typically 5 to 50 nm in diameter, are precursors for the formation of solid and stable nanoparticles. In addition to plasmid DNA, engineered nanoparticulate systems may contain many different materials for various medical and engineering applications such as nanomagnets and nanosensors.The specific goal of the project is to demonstrate that targeted solid nanoparticles containing plasmid DNA can be engineered from novel microemulsion precursors. The project has three specific aims, 1) demonstrate that plasmid DNA can be incorporated into a stable microemulsion and that stable solid nanoparticles (5-50 nm) containing plasmid DNA can be made from the microemulsion precursor, 2) characterize the solid nanoparticles (i.e., size, surface charge and porosity, DNA release and stability) and demonstrate that the solid nanoparticles are stable in biological fluids and can express a transgene in-vitro, and 3) incorporate a cell-specific ligand onto the surface of the solid nanoparticles and demonstrate surface recognition of the targeted nanoparticles.