Martiniuk Frank, Martiniuk Justin, Mack Adra, Cheng Seng H
JME Group Associates, Inc, Roseland, New Jersey, USA.
PsychoGenetics Center, Paramus, New Jersey, USA.
DNA Cell Biol Rep. 2025 May 9;6(1):64-75. doi: 10.1089/dcbr.2025.0025. eCollection 2025 May.
Genetic deficiency of lysosomal acid α-glucosidase or acid maltase (GAA) results in Pompe disease (PD) or glycogen storage disease type II, encompassing subtypes of varying severity. The infantile form presents as hypotonia, muscle weakness, and congestive heart failure in the first year, while the adult onset forms are limited to skeletal muscle. Gene therapy for diseases that affect muscles or organs (as PD) may be very difficult due to their size or location and our ability to transfect these organs with high efficiency.
We evaluated a plasmid vector carrying the cytomegalovirus promoter linked to the human cDNA complexed with the cationic lipid GL-67 as a delivery system for therapy of PD.
A human PD fibroblast cell line was transiently transfected with increasing amounts of the cationic lipid GL-67 complexed to the human expression plasmid. Cells showed a significant increase in human GAA activity with 20 nmol of the cationic lipid, resulting in maximal expression. GAA knockout mice were treated intraperitoneally every 2 weeks for four months with 100 µg of plasmid and 2 mM GL-67 lipid. Tissues, including heart, diaphragm, and skeletal muscle showed substantial increase in GAA activity. More important, the clinical phenotype of hind and forelimb muscle weakness was reversed and life span extended to almost normal longevity.
These data suggest that cationic lipid mediated gene delivery of the human cDNA may be an effective treatment for PD.
溶酶体酸性α-葡萄糖苷酶或酸性麦芽糖酶（GAA）的基因缺陷导致庞贝病（PD）或II型糖原贮积病，包括不同严重程度的亚型。婴儿型在出生后第一年表现为肌张力减退、肌肉无力和充血性心力衰竭，而成人发病型则仅限于骨骼肌。由于肌肉或器官（如PD）的大小或位置以及我们高效转染这些器官的能力，针对影响肌肉或器官的疾病进行基因治疗可能非常困难。
我们评估了一种携带与人类cDNA连接的巨细胞病毒启动子的质粒载体，该质粒载体与阳离子脂质GL-67复合，作为治疗PD的递送系统。
用人表达质粒与阳离子脂质GL-67复合后，以递增剂量对人PD成纤维细胞系进行瞬时转染。当阳离子脂质用量为20 nmol时，细胞显示人GAA活性显著增加，从而实现最大表达。用100μg质粒和2 mM GL-67脂质每2周对GAA基因敲除小鼠进行一次腹腔注射，持续四个月。包括心脏、膈肌和骨骼肌在内的组织显示GAA活性大幅增加。更重要的是，后肢和前肢肌肉无力的临床表型得到逆转，寿命延长至几乎正常寿命。
这些数据表明，阳离子脂质介导的人cDNA基因递送可能是治疗PD的有效方法。
