Human Gene TherapyA developing treatment that aims to fix, replace, or add genetic instructions. 9:1609-1616 (July 20, 1998)
Mary Ann Liebert, Inc.
Adenovirus-Mediated Transfer of Human Acid Maltase Gene
Reduces Glycogen AccumulationThe buildup of glycogen inside cells and tissues when it is not broken down properly. in Skeletal MuscleMuscles that control movement and are commonly affected in Pompe disease. of
Japanese Quail with Acid Maltase Deficiency
S. Tsujino 1), N. Kinoshita 1), T. Tashiro 1), K. Ikeda 1),
N. Ichihara 2), H. Kikuchi 2), Y. Hagiwara 2),
M. Mizutani 2), T. Kikuchi 2), N. Sakuragawa 1)
(1) Departments of Inherited Metabolic Disease and of (2) Animal Models for Human Disease, National Institute of Neuroscience, NCNP, Kodaira, Tokyo, Japan
ABSTRACT
Acid maltase deficiency (AMD)Another name for Pompe disease. causes a lysosomal glycogenosis inherited as an autosomal recessiveA pattern of inheritance where a person must receive one altered gene from each parent to develop Pompe disease. trait. The infantile type of AMD (Pompe diseaseA rare genetic disease in which the body cannot properly break down glycogen, leading to buildup that damages muscles and can affect breathing and, in some cases, the heart.) leads to early death due to severe dysfunction of cardiac and respiratory muscles and no effective therapy is available. Replication-defective adenovirus vectors offer a promising tool for in vivoResearch or testing that is performed in a living organism, such as a person or animal. gene delivery and gene therapy. We constructed a recombinant adenovirus containing 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 animal model for human AMD. When cultured fibroblasts from AMD quail were infected with AxCANAM, AM activity in the cells increased in proportion to the multiplicity of infection (MOI). When AxCANAM (4.5 x 10(8) PFU) was injected into unilateral superficial pectoral muscle of AMD quail, PAS staining showed that glycogenosomes disappeared and stainablility of acid phosphatase was reduced in the injected area as compared with the contralateral muscle of the same birds. Biochemically, AM activity increased and glycogenA stored form of sugar used for energy. content decreased in the injected muscle. Western blot analysis showed that AMD quail muscle injected with AxCANAM expressed human AM protein processed to active forms. These results suggest that the human AM cDNA transferred by an adenovirus vectorA delivery system, often based on a modified virus, used to carry genetic material into cells. was sufficiently expressed, leading to a marked reduction of the glycogen accumulation in the skeletal muscle of AMD quail.