Genome editing for the correction of Pompe disease

基因组编辑用于纠正庞贝病

基本信息

批准号：
10219962
负责人：
Dwight D Koeberl
金额：
$ 20.11万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-07-20 至 2022-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10219962
关键词：
Acids Address Age Alternative Therapies Androgens Biochemical Biological CRISPR/Cas technology Cells Cleaved cell Clustered Regularly Interspaced Short Palindromic Repeats Data Development Disease Disease model Engineering Female Gender Gene Delivery Genes Glucan 1,4-alpha-Glucosidase Glycogen Glycogen storage disease type II Goals Guide RNA Human In Vitro Individual Infant Measures Mediating Medical Modeling Mus Muscle Muscle Cells Muscle Weakness Muscle function Muscular Dystrophies Mutation Myopathy Pathogenicity Patients Positioning Attribute Proteins Publications Rare Diseases Recombinants Reproducibility Research Personnel Skeletal Muscle Staphylococcus aureus Symptoms System Technology Time Tissues Variant adeno-associated viral vector age effect base curative treatments design disease phenotype enzyme activity enzyme replacement therapy experience gene therapy genetic variant genome editing glucosidase homologous recombination improved in vitro Model life time cost metabolic myopathies mouse model mutation correction novel therapeutic intervention personalized medicine repaired response sex stability testing tool transgene delivery vector

项目摘要

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.

庞贝疾病是由溶酶体酸α-缺乏引起的潜在致命代谢肌病葡萄糖酶（GAA; EC 3.2.1.20）导致溶酶体糖原和肌肉损伤的积累。这仅当前可用的治疗包括重组人的酶替代疗法（ERT） GAA，需要经常以高终身成本进行治疗。庞贝疾病中的基因组编辑，相反，可以永久纠正GAA基因中的突变并改善患者症状。这群集定期间隔短的短质体重复序列（CRISPR）-CAS9系统已成为一个具有高精度编辑基因组的有希望的工具。但是，以前使用该系统的研究通过同源重组进行的突变校正需要将每个细胞与两个腺与使用单个AAV矢量AS相比，相对效率相对较低的相关病毒（AAV）向量在这里提议。鉴于AAV矢量不会均匀地翻译肌肉，因此两个载体转导单个肌肉细胞较低。因此，将该技术应用于纠正庞贝疾病的突变是不可能的。我们建议使用单个AAV载体纠正庞贝疾病中的生化异常和肌肉无力。我们将纠正两个最常见的GAA基因变体。庞贝疾病中的基因组编辑将永久纠正GAA并不断表达GAA改善患者符号并代表主要庞贝患者治疗的进步。