Genome editing for the correction of Pompe disease

庞贝氏症是一种潜在的致命性代谢性肌病，由溶酶体酸α- 葡糖苷酶（GAA; EC 3.2.1.20），其导致溶酶体糖原的积累和肌肉损伤。的 目前唯一可用的治疗包括用重组人的酶替代疗法（ERT）， GAA，需要频繁治疗，终身成本高。庞贝氏症中的基因组编辑， 相反，可以永久纠正GAA基因的突变，改善患者的症状。的 CRISPR-Cas9系统已经成为一种新的基因表达系统。 有前途的工具，以高精度编辑基因组。然而，以前的研究，使用这个系统， 通过同源重组的突变校正需要用两个腺病毒转导每个细胞， 与使用单一AAV载体作为载体相比，本发明提供了效率相对较低的AAV相关病毒（AAV）载体。 在这里提出。考虑到AAV载体不均匀地包裹肌肉，两种载体的效率不一致。 载体对单个肌肉细胞的抑制作用很低;因此，应用这项技术来纠正 庞贝氏症的突变是不可能的。我们建议开发一个基因组编辑系统， 在一些实施方案中，使用单一AAV载体来纠正庞贝氏症中的生化异常和肌肉无力。我们将 纠正两种最常见的GAA基因变异。庞贝氏症的基因组编辑将永久性 纠正GAA并持续表达GAA，改善患者症状， Pompe患者的治疗进展。

Pompe disease is a potentially fatal metabolic myopathy caused by the deficiency of lysosomal acid α- glucosidase (GAA; EC 3.2.1.20) that leads to accumulation of lysosomal glycogen and muscle damage. The only currently available treatment consists of enzyme replacement therapy (ERT) with recombinant human GAA, which requires frequent treatments with high lifetime costs. Genome editing in Pompe disease, conversely, could permanently correct mutations in the GAA gene and ameliorate patient symptoms. The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-Cas9 system has emerged as a promising tool to edit genomes with high precision. However, previous studies that used this system for mutation correction through homologous recombination required the transduction of each cell with two adeno- associated virus (AAV) vectors with relatively low efficiency, in comparison with using a single AAV vector as proposed here. Given that AAV vectors do not uniformly transduce muscle, the efficiency with which two vectors transduce individual muscle cells is low; therefore, the application of this technology to correct mutations in Pompe disease has not been possible. We propose to develop a genome editing system using a single AAV vector to correct the biochemical abnormalities and muscle weakness in Pompe disease. We will correct two of the most common GAA gene variants. Genome editing in Pompe disease will permanently correct GAA and continuously express GAA ameliorating patient symptoms and represents a major advancement for the treatment of Pompe patients.