In vitro and in vivo gene therapy with CMV vector-mediated presumed dog beta-nerve growth factor in pyridoxine-induced neuropathy dogs.
Due to the therapeutic potential of gene therapy for neuronal injury, many studies of neurotrophic factors, vectors, and animal models have been performed. The presumed dog beta-nerve growth factor (pdbeta-NGF) was generated and cloned and its expression was confirmed in CHO cells. The recombinant pdbeta-NGF protein reacted with a human beta-NGF antibody and showed bioactivity in PC12 cells. The pdbeta-NGF was shown to have similar bioactivity to the dog beta-NGF. The recombinant pdbeta-NGF plasmid was administrated into the intrathecal space in the gene therapy group. Twenty-four hours after the vector inoculation, the gene therapy group and the positive control group were intoxicated with excess pyridoxine for seven days. Each morning throughout the test period, the dogs' body weight was taken and postural reaction assessments were made. Electrophysiological recordings were performed twice, once before the experiment and once after the test period. After the experimental period, histological analysis was performed. Dogs in the gene therapy group had no weight change and were normal in postural reaction assessments. Electrophysiological recordings were also normal for the gene therapy group. Histological analysis showed that neither the axons nor the myelin of the dorsal funiculus of L4 were severely damaged in the gene therapy group. In addition, the dorsal root ganglia of L4 and the peripheral nerves (sciatic nerve) did not experience severe degenerative changes in the gene therapy group. This study is the first to show the protective effect of NGF gene therapy in a dog model.
Chung
Jin Young
JY
Department of Veterinary Internal Medicine, College of Veterinary Medicine, Seoul National University, Seoul 151-742, Korea.
Choi
Jung Hoon
JH
Shin
Il Seob
IS
Choi
Eun Wha
EW
Hwang
Cheol Yong
CY
Lee
Sang Koo
SK
Youn
Hwa Young
HY
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D016466:CHO Cells; D002493:Central Nervous System Diseases / Q000139:chemically induced / Q000628:therapy / Q000662:veterinary*; D003001:Cloning, Molecular; D006224:Cricetinae; D003412:Cricetulus; D003587:Cytomegalovirus; D004283:Dog Diseases / Q000139:chemically induced* / Q000628:therapy; D004285:Dogs; D005260:Female; D015316:Genetic Therapy / Q000662:veterinary*; D005822:Genetic Vectors; D008297:Male; D008969:Molecular Sequence Data; D020932:Nerve Growth Factor / Q000235:genetics / Q000378:metabolism* / Q000627:therapeutic use*; D011736:Pyridoxine / Q000633:toxicity*
D020932:Nerve Growth Factor; D011736:Pyridoxine