Primary astrocytes retrovirally transduced with a tyrosine hydroxylase transgene driven by a glial-specific promoter elicit behavioral recovery in experimental parkinsonism.
We used a retroviral-mediated gene transfer system to transduce primary rat astrocytes with a transgene in which the activity of a tyrosine hydroxylase (TH) cDNA is under the transcriptional control of a human promoter of the glial fibrillary acidic protein (GFAP). The engineered cells were tested for their therapeutic efficacy in a rodent model of Parkinson's disease (PD). The method is based both on the properties of astrocytes, as well as on those of the promoter. Astrocytes are an integral part of the neural tissue, have a long life span, are more resistant to oxidative stress than neurons, and possess an efficient secretory system. The GFAP promoter is active throughout postnatal life, and its activity is up-regulated by many insults to the brain, including PD. Transduced astrocytes were implanted into the striata of rats lesioned with 6-hydroxydopamine (6-OHDA), and the efficacy of grafted cells tested. Implanted astrocytes induced a significant reduction in the turning behavior that occurs in response to apomorphine for at least 4 weeks after grafting, and transgenic mRNA and protein could be detected in implanted brains. These results indicate that the gFa2-TH construct can be readily adapted to be used with a retroviral gene transfer system to obtain nontumorigenic cells that sustain a sufficient level of transgene activity to enable therapeutic effectiveness for prolonged periods. These results further endorse the use of astrocytes for gene therapy in the central nervous system.
Cortez
N
N
Departamento de Fisiología, Biofísica y Neurociencias Centro de Investigación y de Estudios Avanzados del IPN, México, DF.
Trejo
F
F
Vergara
P
P
Segovia
J
J
D000818:Animals; D001253:Astrocytes / Q000201:enzymology* / Q000821:virology*; D018929:Cell Culture Techniques; D005904:Glial Fibrillary Acidic Protein / Q000235:genetics; D007150:Immunohistochemistry; D008297:Male; D020734:Parkinsonian Disorders / Q000235:genetics / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D051381:Rats; D017208:Rats, Wistar; D012190:Retroviridae / Q000235:genetics*; D019076:Transgenes / Q000235:genetics*; D014446:Tyrosine 3-Monooxygenase / Q000235:genetics*
D005904:Glial Fibrillary Acidic Protein; D014446:Tyrosine 3-Monooxygenase