Intrastriatal grafts of embryonic mesencephalic rat neurons genetically modified using an adenovirus encoding human Cu/Zn superoxide dismutase.
Intrastriatal grafting of embryonic dopamine-containing neurons is a promising approach for treating clinical and experimental Parkinson's disease. However, neuropathological analyses of grafted patients and transplanted rats have demonstrated that the survival of grafted dopamine neurons is relatively poor. In the present study, we pursued a strategy of transferring a potentially neuroprotective gene into rat embryonic mesencephalic rat cells in vitro, before grafting them into the denervated striatum of 6-hydroxydopamine-lesioned rats. We performed intrastriatal grafts of embryonic day 14 mesencephalic cells infected with replication-defective adenoviruses bearing either the human copper-zinc superoxide dismutase gene or, as a control, the E. coli lac Z marker gene. The transgenes were expressed in the grafts four days after transplantation and the expression persisted for at least five weeks thereafter. After five weeks postgrafting, there was more extensive functional recovery in the superoxide dismutase group as compared to the control (uninfected cells) and beta-galactosidase groups. The functional recovery was significantly correlated with the number of tyrosine hydroxylase-positive cells in the grafts, although the clear trend to increased survival of the dopamine neurons in the superoxide dismutase grafts did not reach statistical significance. Only a moderate inflammatory reaction was revealed by OX-42 immunostaining in all groups, suggesting that ex vivo gene transfer using adenoviral vectors is a promising method for delivering functional proteins into brain grafts.
Barkats
M
M
Laboratoire de Génétique Moléculaire de la Neurotransmission et des Processus Neurodégénératifs, UMR CNRS C9923, Hôpital de la Pitié Salpêtrière, Paris, France.
Nakao
N
N
Grasbon-Frodl
E M
EM
Bilang-Bleuel
A
A
Revah
F
F
Mallet
J
J
Brundin
P
P
D000818:Animals; D001358:Avian Sarcoma Viruses / Q000235:genetics; D016380:Brain Tissue Transplantation / Q000502:physiology*; D017690:Cell Transplantation / Q000502:physiology*; D004298:Dopamine / Q000502:physiology; D005260:Female; D016332:Fetal Tissue Transplantation / Q000502:physiology*; D005822:Genetic Vectors; D006085:Graft Survival; D006801:Humans; D007150:Immunohistochemistry; D007786:Lactose Factors / Q000235:genetics; D017072:Neostriatum / Q000637:transplantation*; D009474:Neurons / Q000378:metabolism* / Q000502:physiology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013482:Superoxide Dismutase / Q000378:metabolism*; D014446:Tyrosine 3-Monooxygenase / Q000378:metabolism; D001616:beta-Galactosidase / Q000378:metabolism
D014446:Tyrosine 3-Monooxygenase; D013482:Superoxide Dismutase; D001616:beta-Galactosidase; D004298:Dopamine