Fraga Michelle, de Carvalho Talita Giacomet, Diel Dirnete da Silva, Kretzmann Filho Nélson Alexandre, Teixeira Helder Ferreira, Matte Ursula
J Nanosci Nanotechnol. 2015 Jan;15(1):810-6. doi: 10.1166/jnn.2015.9179.
Mucopolysaccharidosis I (MPS I) is an autosomal recessive lysosomal storage disease due to deficient a-L-iduronidase (IDUA) activity. It results in the accumulation of the glycosaminoglycans (GAGs) heparan and dermatan sulfate and leads to several clinical manifestations. This study describes the use of cationic nanoemulsions as a non-viral carrier for the plasmid named pIDUA, which has the gene that encodes for the IDUA enzyme. Cationic nanoemulsions, composed by a medium chain triglycerides oil core stabilized by DOTAP, DOPE and DSPE-PEG, were prepared by high pressure homogeneization. pIDUA was complexed with nanoemulsions in the end of manufacturing process. Physicochemical properties of complexes were influenced by the charge ratio used. From a charge ratio of +2/-, it was observed a total complexation of pIDUA with formulation as well as a protection of plasmid against DNAse I digestion. In vitro assay in fibroblasts of one MPS I patient presented greater and significant trasfection efficiency for pIDUA complexed to formulation in the +4/- charge ratio. This formulation was administered via the tail vein and the portal vein. Animals were compared to untreated MPS I mice. Transfection efficiency was measured as IDUA enzyme activity. After intravenous administration, IDUA activity was significantly higher in lungs and liver. The set of results shows the formulation obtained at the +4/- charge ratio as a promising non-viral gene delivery system, once showed increased enzyme activity both in vitro and in vivo.
黏多糖贮积症 I 型（MPS I）是一种常染色体隐性溶酶体贮积病，因α-L-艾杜糖醛酸酶（IDUA）活性缺乏所致。它导致糖胺聚糖（GAGs）硫酸乙酰肝素和硫酸皮肤素蓄积，并引发多种临床表现。本研究描述了使用阳离子纳米乳剂作为名为 pIDUA 的质粒的非病毒载体，该质粒含有编码 IDUA 酶的基因。由中链甘油三酯油核组成、由 DOTAP、DOPE 和 DSPE-PEG 稳定的阳离子纳米乳剂通过高压均质法制备。在制造过程结束时，将 pIDUA 与纳米乳剂复合。复合物的物理化学性质受所用电荷比的影响。从 +2/- 的电荷比开始，观察到 pIDUA 与制剂完全复合，以及质粒对 DNA 酶 I 消化的保护作用。在一名 MPS I 患者的成纤维细胞中进行的体外试验表明，与制剂复合的 pIDUA 在 +4/- 电荷比下具有更高且显著的转染效率。该制剂通过尾静脉和门静脉给药。将动物与未治疗的 MPS I 小鼠进行比较。转染效率以 IDUA 酶活性来衡量。静脉给药后，肺和肝中的 IDUA 活性显著更高。这组结果表明，以 +4/- 电荷比获得的制剂是一种有前景的非病毒基因递送系统，因为它在体外和体内均显示出酶活性增加。
