Dopa-producing astrocytes generated by adenoviral transduction of human tyrosine hydroxylase gene: in vitro study and transplantation to hemiparkinsonian model rats.
Astrocytes secreting a large amount of 3,4-dihydroxyphenylalanine (dopa) were generated by adenoviral transduction of the human tyrosine hydroxylase (TH) gene. After characterizing in vitro, the effect of transplantation of these astrocytes to the striatum of hemiparkinsonian model rats was investigated. Subconfluent cortical astrocytes were infected by replication-defect adenovirus type 5 carrying the human TH-1 gene or the LacZ reporter gene under the promoter of the glial fibrillary acidic protein (AdexGFAP-HTH-1, AdexGFAP-NL-LacZ). Dopa secretion was not evident at 3 days after the transduction of the HTH-1 gene but it increased from 7 days up to at least 4 months. The secretion was substrate (tyrosine)-dependent, and was enhanced by loading tetrahydrobioputerin (BH4) concentration-dependently. One-third of the hemiparkinsonian model rats, that were transplanted the HTH-1 gene-transduced astrocytes or introduced the direct injection of the viral vector to the striatum, showed a reduction of methamphetamine-induced rotations for at least 6 weeks. Apomorphine-induced rotation was decreased to the 50% level of the control's, but the reduction was obtained equally by the transplantation of HTH-1 gene-transduced or LacZ reporter gene-transduced astrocytes, or by the introduction of HTH-1 or LacZ gene carrying adenovirus. Treatment with FK506 for 3 weeks improved the late-phase apomorphine-induced rotations following the introduction of the HTH-1 gene carrying adenovirus. Histological examination revealed that, in animals that showed a reduction of methamphetamine-rotation, the TH positive astrocytes-like cells were distributed widely in the host striatum for at least 4 weeks. The number of TH positive astrocytes-like cells and their immunoreactivity decreased after 6 weeks when OX-41 positive microglias/macrophages were infiltrated. Data indicate that the adenoviral transduction of the human TH gene to astrocytes and its introduction to the striatum is a promising approach for the treatment of Parkinson's disease. However, the further technical improvements are required to optimize the adenoviral gene delivery, such as the control of viral toxicity and the regulation of the immune response.
Hida
H
H
Department of Physiology, Nagoya City University Medical School, Nagoya, Japan.
Hashimoto
M
M
Fujimoto
I
I
Nakajima
K
K
Shimano
Y
Y
Nagatsu
T
T
Mikoshiba
K
K
Nishino
H
H
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D000831:Animals, Newborn; D001058:Apomorphine; D001253:Astrocytes / Q000166:cytology / Q000201:enzymology / Q000637:transplantation* / Q000821:virology; D002478:Cells, Cultured; D003342:Corpus Striatum / Q000201:enzymology / Q000473:pathology / Q000821:virology; D004295:Dihydroxyphenylalanine / Q000096:biosynthesis* / Q000235:genetics; D005260:Female; D005822:Genetic Vectors / Q000235:genetics; D006084:Graft Rejection; D006801:Humans; D007166:Immunosuppressive Agents / Q000494:pharmacology; D008694:Methamphetamine; D051379:Mice; D008813:Mice, Inbred ICR; D010300:Parkinson Disease / Q000201:enzymology / Q000473:pathology / Q000628:therapy*; D004856:Postural Balance / Q000187:drug effects; D051381:Rats; D017208:Rats, Wistar; D012678:Sensation Disorders / Q000139:chemically induced / Q000517:prevention & control; D016559:Tacrolimus / Q000494:pharmacology; D014162:Transfection; D014446:Tyrosine 3-Monooxygenase / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
D007166:Immunosuppressive Agents; D008694:Methamphetamine; D004295:Dihydroxyphenylalanine; D014446:Tyrosine 3-Monooxygenase; D001616:beta-Galactosidase; D001058:Apomorphine; D016559:Tacrolimus