MuSC dysfunction contributes to dystrophic progression and impaired regeneration in the mdx mouse [bulk RNA-seq]

Duchenne muscular dystrophy (DMD) is a devastating disease characterized by progressive muscle wasting and limitation of life. In addition to the inherent weakness of dystrophin-deficient muscle, the dysfunction of resident muscle stem cells significantly contributes to disease progression. Using the mdx mouse model of DMD, we performed an in-depth characterization of disease progression and muscle stem cell (MuSC) function in dystrophin deficient skeletal muscle using immunohistology, isometric force measurements, transplantation assays, and transcriptomic analysis. Here, we performed bulk RNA-sequencing of wild-type (WT) and mdx myogenic cells in different conditions. Overall design: Bulk RNA-sequencing of mogenic cells derived from WT and mdx Pax7-nGFP mice was performed in biological triplicate for 4 conditions (24 libraries total). PAX7 expressing MuSCs were isolated from hindlimb muscles before and 3-days after cardiotoxin injury. Myoblasts and 2-day differentiated myotubes were also collected after in vitro culture of WT and mdx PAX7 expressing cells.

杜氏肌营养不良症（Duchenne muscular dystrophy, DMD）是一种以进行性肌肉萎缩和寿命受限为特征的毁灭性疾病。除肌营养不良蛋白缺陷型肌肉本身的固有脆弱性外，驻留肌肉干细胞（muscle stem cell, MuSC）的功能异常显著推动了疾病进展。本研究利用DMD的mdx小鼠模型，通过免疫组织化学（immunohistology）、等长肌力测量（isometric force measurements）、移植实验（transplantation assays）及转录组分析（transcriptomic analysis），对肌营养不良蛋白缺陷型骨骼肌的疾病进展与肌肉干细胞功能开展了深入表征。本研究对不同条件下的野生型（wild-type, WT）及mdx肌源性细胞进行了批量RNA测序（bulk RNA-sequencing）。整体实验设计：针对4种实验条件，分别采集来自野生型及mdx Pax7-nGFP小鼠的肌源性细胞样本，设置3次生物学重复，共计24个文库。具体分组如下：在心脏毒素损伤前及损伤3天后，从后腿肌肉中分离表达PAX7的肌肉干细胞；同时，对体外培养的野生型及mdx PAX7阳性细胞，分别收集成肌细胞以及诱导分化2天的肌管样本。