Melo Dyanna, Maruyama Rika, Yokota Toshifumi
Department of Medical Genetics, Faculty of Medicine and Dentistry, University of Alberta, Edmonton, AB, Canada.
The Friends of Garrett Cumming Research & Muscular Dystrophy Canada HM Toupin Neurological Science Research Chair, Edmonton, AB, Canada.
Methods Mol Biol. 2025;2964:231-242. doi: 10.1007/978-1-0716-4730-1_15.
Duchenne muscular dystrophy (DMD) is an X-linked recessive disorder due to the lack of dystrophin production. The disease is characterized by muscle wasting, with the most common causes of death being respiratory failure or heart failure. Recently, exon skipping via phosphorodiamidate morpholino oligomers (PMOs) are used as an FDA approved treatment for DMD. Peptide-conjugated PMOs (PPMOs) are used to increase exon skipping efficacy in the heart and are a promising therapy for DMD. Researchers have previously relied on high-performance liquid chromatography (HPLC) or liquid chromatography-mass spectrometry (LC/MS) methods for detecting PPMO uptake, but an enzyme-linked immunosorbent assay (ELISA) has been shown to have greater sensitivity. Here, we present methodologies to determine the uptake efficiency of a PPMO into the heart and efficacy of exon 51 skipping by a PPMO injected retro-orbitally into a humanized DMD mouse model via ELISA and RT-PCR, respectively.
杜氏肌营养不良症（DMD）是一种由于缺乏抗肌萎缩蛋白产生而导致的X连锁隐性疾病。该疾病的特征是肌肉萎缩，最常见的死亡原因是呼吸衰竭或心力衰竭。最近，通过磷酰二胺吗啉代寡聚物（PMO）进行的外显子跳跃被用作美国食品药品监督管理局（FDA）批准的DMD治疗方法。肽缀合的PMO（PPMO）用于提高心脏中外显子跳跃的效率，是一种有前景的DMD治疗方法。研究人员此前一直依赖高效液相色谱（HPLC）或液相色谱-质谱联用（LC/MS）方法来检测PPMO的摄取情况，但已证明酶联免疫吸附测定（ELISA）具有更高的灵敏度。在此，我们分别通过ELISA和RT-PCR展示了确定PPMO进入心脏的摄取效率以及通过眼眶后注射到人性化DMD小鼠模型中的PPMO进行外显子51跳跃的功效的方法。
