Gene delivery to hair follicles by multifunctional envelope-type nano device

本项目的主要研究人员试图通过局部应用非病毒基因递送系统八精氨酸修饰的多功能包膜型纳米器件(R8-MEND)来开发新型的小鼠毛囊基因递送系统。用含EGFP编码质粒DNA的R8-MEND和编码LacZ的质粒DNA处理小鼠背部皮肤后，在R8-MEND处理的皮肤切片上观察到EGFP蛋白的表达和β-Gal阳性染色图像。此外，将含有骨形态发生蛋白IA受体(BMPRIA)编码质粒DNA的R8-MEND局部应用于4周龄ICR小鼠的背部。治疗2周后，毛囊在皮肤中的位置与治疗前基本相同，但对照组小鼠毛囊位置因毛发周期的推进而上移。建议外用R8-MEND-BMPRIA延长毛发周期。然后，首席调查员t…更努力地抑制与头发生长和周期相关的特定基因，方法是通过局部应用R8-MEND来针对这些基因传递siRNA。然而，没有观察到明显的效果。为了了解R8-MEND传递siRNA无效的原因，首席调查员观察了R8-MEND处理的皮肤横断面，其中含有罗丹明标记的siRNA。因此，在皮肤中观察到少量的siRNA，这表明有必要改进siRNA的输送。然后，首席研究人员试图通过离子导入来传递脂质体，这是MEND的模型。首席研究人员发现，通过离子导入，脂质体的最佳尺寸(&gt；400 nm)和表面电荷(正电荷)可以通过卵泡传递纳米颗粒。此外，首席研究员通过体内离子导入成功地将含有模型药物溶液(罗丹明水)的脂质体通过毛囊输送到皮肤。因此，这种新型的跨叶递送系统有望在不久的将来实现含有siRNA的R8-MEND的递送。较少

Head investigator of this project tried to develop novel gene delivery system to hair follicles of mice by topical application of non-viral gene delivery system octaarginine-modified multifunctional envelope-type nano device (R8-MEND). After treatment of mice back skin with the R8-MEND containing EGFP-encoding plasmid DNA or LacZ-encoding plasmid DNA, expression of EGFP protein and beta-gal positive staining image were observed in cross section of the skin treated with R8-MEND. Moreover, topical application of R8-MEND, which contained Bone morphogenetic protein type IA receptor (BMPRIA) -encoding plasmid DNA, was performed to back skin of 4 weeks old ICR mice. After 2 weeks of the treatment, position of hair follicles in the skin was almost the same as before treatment, although position of hair follicles of control mice was moved to upper region because of progress of hair cycle. It is suggested that topical application of R8-MEND-BMPRIA prolonged hair cycle. Then, head investigator t … More ried to inhibit the specific genes relating to growth and cycle of hair by delivery of siRNA against those genes via topical application of R8-MEND. However, no significant effect was observed. To know the reason why the siRNA delivery by R8-MEND was not effective, head investigator observed cross section of the skin treated by R8-MEND containing siRNA labeled with rhodamine. As a result, very a few siRNA was observed in the skin, indicating that improvement of siRNA delivery is necessary. Then, head investigator tried to delivery liposomes, which are the model of MEND, via iontophoresis. Head investigator found optimal size (>400nm) and surface charge (positive) of liposomes for transfollicular delivery of nano-particles via iontophoresis. Moreover, head investigator succeeded in transfollicular delivery of liposomes containing model drug solution (aqueous rhodamine) into skin via iontophoresis in vivo. Therefore, the delivery of R8-MEND containing siRNA should be achieved by this novel transfolicular delivery system in near future. Less