FVIII gene delivery by muscle electroporation corrects murine hemophilia A

• Published in: The journal of gene medicine 2005-04, Vol.7 (4), p.494-505
• Main Authors: Long, Yun Chau, Jaichandran, S., Ho, Liam Pock, Tien, Sim Leng, Tan, Soo Yong, Kon, Oi Lian
• Format: Article
• Language: English
• Subjects: Animals; Base Sequence; Cell Line; Culture Media, Conditioned; DNA Primers; Electroporation; factor VIII; Factor VIII - genetics; Gene therapy; Genes, Reporter; Genetic Therapy; hemophilia; Hemophilia A - therapy; Mice; muscle; Muscle, Skeletal - metabolism; Phenotype; plasmid; Plasmids
• ISSN: 1099-498X; 1521-2254
• DOI: 10.1002/jgm.683

Background Hemophilia A treatment relies on costly factor VIII (FVIII) replacement that may transmit iatrogenic viral diseases. Viral vectors and cell implants are being developed as improvements. We investigated in vivo electroporation of naked DNA as a safe and simple method for correcting FVIII deficiency. Methods B‐domain‐deleted murine FVIII cDNA expression plasmids were constructed with CMV and elongation factor 1α promoters for characterisation in murine C2C12 myoblasts. The construct conferring highest in vitro FVIII secretion was electroporated into skeletal muscle of FVII null mice in vivo for phenotypic correction using a protocol that minimised tissue injury. Results B‐domain‐deleted murine FVIII cDNA plasmids induced FVIII secretion from stably transfected C2C12 myoblasts (0.54 ± 0.20 mU/day/105 cells). Phenotypic correction of hemophilic mice was more consistently achieved using a protocol for in vivo electroporation of gastrocnemius muscle with FVIII cDNA that reduced tissue injury by the use of plate electrodes, hyaluronidase pre‐treatment and lower field strength. This technique was associated with