Sekine Shohko, Mayama Sayaka, Nishijima Nobuaki, Kojima Takuo, Endo-Takahashi Yoko, Ishii Yuko, Shiono Hitomi, Akiyama Saki, Sakurai Akane, Sashida Sanae, Hamano Nobuhito, Tada Rui, Suzuki Ryo, Maruyama Kazuo, Negishi Yoichi
Department of Drug Delivery and Molecular Biopharmaceutics, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, Tokyo 192-0392, Japan.
Laboratory of Drug and Gene Delivery Research, Faculty of Pharma-Sciences, Teikyo University, 2-11-1 Kaga, Itabashi-ku, Tokyo 173-8605, Japan.
Pharmaceutics. 2023 Jun 6;15(6):1665. doi: 10.3390/pharmaceutics15061665.
Strategies for gene and nucleic acid delivery to skeletal muscles have been extensively explored to treat Duchenne muscular dystrophy (DMD) and other neuromuscular diseases. Of these, effective intravascular delivery of naked plasmid DNA (pDNA) and nucleic acids into muscles is an attractive approach, given the high capillary density in close contact with myofibers. We developed lipid-based nanobubbles (NBs) using polyethylene-glycol-modified liposomes and an echo-contrast gas and found that these NBs could improve tissue permeability by ultrasound (US)-induced cavitation. Herein, we delivered naked pDNA or antisense phosphorodiamidate morpholino oligomers (PMOs) into the regional hindlimb muscle via limb perfusion using NBs and US exposure. pDNA encoding the luciferase gene was injected with NBs via limb perfusion into normal mice with application of US. High luciferase activity was achieved in a wide area of the limb muscle. DMD model mice were administered PMOs, designed to skip the mutated exon 23 of the dystrophin gene, with NBs via intravenous limb perfusion, followed by US exposure. The number of dystrophin-positive fibers increased in the muscles of mdx mice. Combining NBs and US exposure, which can be widely delivered to the hind limb muscles via the limb vein, could be an effective therapeutic approach for DMD and other neuromuscular disorders.
为治疗杜氏肌营养不良症（DMD）和其他神经肌肉疾病，人们已广泛探索了将基因和核酸递送至骨骼肌的策略。其中，鉴于与肌纤维紧密接触的高毛细血管密度，将裸露的质粒DNA（pDNA）和核酸有效血管内递送至肌肉是一种有吸引力的方法。我们使用聚乙二醇修饰的脂质体和一种超声造影气体开发了基于脂质的纳米气泡（NBs），并发现这些NBs可通过超声（US）诱导的空化作用提高组织通透性。在此，我们通过使用NBs和US照射的肢体灌注，将裸露的pDNA或反义磷酰胺吗啉代寡聚物（PMOs）递送至后肢局部肌肉。编码荧光素酶基因的pDNA通过肢体灌注与NBs一起注射到正常小鼠体内，并施加US。在肢体肌肉的广泛区域实现了高荧光素酶活性。DMD模型小鼠通过静脉肢体灌注接受与NBs一起给药的PMOs，这些PMOs旨在跳过肌营养不良蛋白基因的突变外显子23，随后进行US照射。mdx小鼠肌肉中抗肌萎缩蛋白阳性纤维的数量增加。将NBs和US照射相结合，可通过肢体静脉广泛递送至后肢肌肉，可能是治疗DMD和其他神经肌肉疾病的有效治疗方法。
