Gene therapy with secreted acid alpha-glucosidase rescues Pompe disease in a novel mouse model with early-onset spinal cord and respiratory defects. Gene therapy with secreted acid alpha-glucosidase rescues Pompe disease in a novel mouse model with early-onset spinal cord and respiratory defects

Pompe disease (PD) is a neuromuscular disorder caused by deficiency of acid-alpha-glucosidase (GAA), leading to motor and respiratory dysfunctions. Available Gaa knock-out (KO) mouse models do not accurately mimic PD, particularly the highly impaired respiratory phenotype. Methods. Here we developed a new mouse model of PD crossing Gaa KO B6;129 with DBA2/J mice. Findings. Male Gaa KODBA2/J presents most of the key features of the human disease, including early lethality, severe respiratory impairment, cardiac hypertrophy and muscle weakness. Transcriptome analyses of Gaa KODBA2/J, compared to the parental Gaa KOB6;129 mice, revealed a profoundly impaired gene signature in the spinal cord and a similarly deregulated gene expression in skeletal muscle. Muscle and spinal cord transcriptome changes in Gaa KODBA2/J, were significantly improved upon gene therapy with AAV vectors expressing a secreted GAA enzyme. Overall design: Transcriptomic analysis comparing the gene expression profiles of quadriceps muscle and whole spinal cord from: Gaa WT and Gaa KO mice in B6;129 background; Gaa WT and Gaa KO mice in B6;129;DBA2/J background and Gaa KO mice in B6;129;DBA2/J strain treated with AAV-mediated gene therapy with secreted acid-alpha-glucosidase.

庞贝病（Pompe disease, PD）是因酸性α-葡萄糖苷酶（acid-alpha-glucosidase, GAA）缺乏引发的神经肌肉疾病，可导致运动与呼吸功能障碍。现有Gaa基因敲除（knock-out, KO）小鼠模型无法精准模拟庞贝病，尤其是重度呼吸功能受损的表型。方法：本研究构建新型庞贝病小鼠模型，通过将Gaa KO B6;129小鼠与DBA2/J小鼠杂交获得。研究结果：雄性Gaa KO DBA2/J小鼠展现出人类庞贝病的多数关键特征，包括早期致死、严重呼吸功能损伤、心肌肥厚与肌肉无力。对Gaa KO DBA2/J小鼠与亲代Gaa KO B6;129小鼠开展转录组分析发现，其脊髓内基因表达特征受到显著破坏，骨骼肌中的基因表达同样存在显著失调。针对Gaa KO DBA2/J小鼠施以表达分泌型GAA酶的腺相关病毒（adeno-associated virus, AAV）载体基因治疗后，其肌肉与脊髓的转录组变化得到显著改善。整体实验设计：对以下样本的股四头肌与全脊髓进行基因表达谱转录组分析：1. B6;129背景下的Gaa野生型（wild-type, WT）与Gaa KO小鼠；2. B6;129;DBA2/J背景下的Gaa WT与Gaa KO小鼠；3. 经AAV介导的分泌型酸性α-葡萄糖苷酶基因治疗后的B6;129;DBA2/J背景Gaa KO小鼠。