Negishi Yoichi, Ishii Yuko, Nirasawa Kei, Sasaki Eri, Endo-Takahashi Yoko, Suzuki Ryo, Maruyama Kazuo
Department of Drug Delivery and Molecular Biopharmaceutics, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, 1432-1 Horinouchi, Hachioji, 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.
Methods Mol Biol. 2018;1687:185-192. doi: 10.1007/978-1-4939-7374-3_13.
Duchenne muscular dystrophy (DMD) is a genetic disorder characterized by progressive muscle degeneration, caused by nonsense or frameshift mutations in the dystrophin (DMD) gene. Antisense oligonucleotides can be used to induce specific exon skipping; recently, a phosphorodiamidate morpholino oligomer (PMO) has been approved for clinical use in DMD. However, an efficient PMO delivery strategy is required to improve the therapeutic efficacy in DMD patients. We previously developed polyethylene glycol (PEG)-modified liposomes containing ultrasound contrast gas, "Bubble liposomes" (BLs), and found that the combination of BLs with ultrasound exposure is a useful gene delivery tool. Here, we describe an efficient PMO delivery strategy using the combination of BLs and ultrasound exposure to treat muscles in a DMD mouse model (mdx). This ultrasound-mediated BL technique can increase the PMO-mediated exon-skipping efficiency, leading to significantly increased dystrophin expression. Thus, the combination of BLs and ultrasound exposure may be a feasible PMO delivery method to improve therapeutic efficacy and reduce the PMO dosage for DMD treatment.
杜氏肌营养不良症（DMD）是一种遗传性疾病，其特征为进行性肌肉退化，由肌营养不良蛋白（DMD）基因中的无义或移码突变引起。反义寡核苷酸可用于诱导特定外显子跳跃；最近，一种磷酰二胺吗啉代寡聚物（PMO）已被批准用于DMD的临床治疗。然而，需要一种有效的PMO递送策略来提高DMD患者的治疗效果。我们之前开发了含有超声造影气体的聚乙二醇（PEG）修饰脂质体，即“气泡脂质体”（BLs），并发现BLs与超声照射相结合是一种有用的基因递送工具。在此，我们描述了一种使用BLs与超声照射相结合的有效PMO递送策略，用于治疗DMD小鼠模型（mdx）的肌肉。这种超声介导的BL技术可以提高PMO介导的外显子跳跃效率，从而显著增加肌营养不良蛋白的表达。因此，BLs与超声照射相结合可能是一种可行的PMO递送方法，可提高治疗效果并减少DMD治疗所需的PMO剂量。
