ADENOVIRUS-MEDIATED TRANSFER OF HUMAN ACID MALTASE GENE REDUCES GLYCOGEN ACCUMULATION IN SKELETAL-MUSCLE OF JAPANESE-QUAIL WITH ACID MALTASE DEFICIENCY

Acid maltase deficiency (AMD) causes a lysosomal glycogenosis inherite d as an autosomal recessive trait. The infantile type of AMD (Pompe di sease) leads to early death due to severe dysfunction of cardiac and r espiratory muscles and no effective therapy is available. Replication- defective adenovirus vectors offer a promising tool for in vivo gene d elivery and gene therapy. We constructed a recombinant adenovirus cont aining the human acid maltase (AM) cDNA downstream of the CAG promoter , composed of modified chicken beta-actin promoter and CMV IE enhancer (AxCANAM), Japanese quail with AMD was used for this study as an anim al model for human AMD, When cultured fibroblasts from AMD quail were infected with AxCANAM, AM activity in the cells increased in proportio n to the multiplicity of infection (MOI), When AxCANAM (4.5 x 10(8) PF U) was injected into unilateral superficial pectoral muscle of AMD qua il, PAS staining showed that glycogenosomes disappeared and stainabili ty of acid phosphatase was reduced in the injected area as compared wi th the contralateral muscle of the same birds, Biochemically, AM activ ity increased and glycogen content decreased in the injected muscle. W estern blot analysis showed that AMD quail muscle injected with AxCANA M expressed human AM protein processed to active forms, These results suggest that the human AM cDNA transferred by an adenovirus vector was sufficiently expressed, leading to a marked reduction of the glycogen accumulation in the skeletal muscle of AMD quail.