Efficient and stable transduction of dopaminergic neurons in rat substantia nigra by rAAV 2 1, 2 2, 2 5, 2 6.2, 2 7, 2 8 and 2 9

• Published in: Gene therapy Vol. 18; no. 5; pp. 517 - 527
• Main Authors: Van der Perren, A, Toelen, J, Carlon, M, Van den Haute, C, Coun, F, Heeman, B, Reumers, V, Vandenberghe, L H, Wilson, J M, Debyser, Z, Baekelandt, V
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 01.05.2011; Nature Publishing Group
• Subjects: 631/1647/2300/1850; 631/326/596/2561; 692/698/1688/64; 692/699/375/346/1718; Anesthesia. Intensive care medicine. Transfusions. Cell therapy and gene therapy; Animal models; Animals; Applied cell therapy and gene therapy; Biological and medical sciences; Bioluminescence; Biomedical and Life Sciences; Biomedicine; Biotechnology; Care and treatment; Cell Biology; Central nervous system; Dependovirus - genetics; Dopamine; Dopamine - metabolism; Dopamine receptors; Expression vectors; Fundamental and applied biological sciences. Psychology; Gene Expression; Gene Therapy; Gene transfer; Genetic aspects; Genetic Vectors; Green Fluorescent Proteins - genetics; Green Fluorescent Proteins - metabolism; Health aspects; Health. Pharmaceutical industry; Human Genetics; Immunohistochemistry; Industrial applications and implications. Economical aspects; Injection; Medical sciences; Movement disorders; Nanotechnology; Neurodegenerative diseases; Neuroimaging; Neurons; original-article; Parkinson's disease; Physiological aspects; Rats; Rats, Wistar; Serotypes; Serotyping; Substantia nigra; Substantia Nigra - cytology; Substantia Nigra - metabolism; Transduction; Transduction, Genetic; Transfusions. Complications. Transfusion reactions. Cell and gene therapy; Tropism; Belgium; adeno-associated viral vectors; bioluminescence imaging; substantia nigra; Parkinson's disease; dopaminergic neurons; Bioluminescence; Rat; Parkinson disease; in vivo bioluminescence imaging; Imaging; Degenerative disease; Nervous system diseases; Rodentia; Dependovirus; Cerebral disorder; Virus; In vivo; Vertebrata; Mammalia; Neuron; Parvoviridae; Central nervous system disease; Transduction; Gene therapy; Parvovirinae; Extrapyramidal syndrome
• ISSN: 0969-7128; 1476-5462
• DOI: 10.1038/gt.2010.179

Dysfunction of the nigrostriatal system is the major cause of Parkinson's disease (PD). This brain region is therefore an important target for gene delivery aiming at disease modeling and gene therapy. Recombinant adeno-associated viral (rAAV) vectors have been developed as efficient vehicles for gene transfer into the central nervous system. Recently, several serotypes have been described, with varying tropism for brain transduction. In light of the further development of a viral vector-mediated rat model for PD, we performed a comprehensive comparison of the transduction and tropism for dopaminergic neurons (DNs) in the adult Wistar rat substantia nigra (SN) of seven rAAV vector serotypes (rAAV 2/1, 2/2, 2/5, 2/6.2, 2/7, 2/8 and 2/9). All vectors were normalized by titer and volume, and stereotactically injected into the SN. Gene expression was assessed non-invasively and quantitatively in vivo by bioluminescence imaging at 2 and 5 weeks after injection, and was found to be stable over time. Immunohistochemistry at 6 weeks following injection revealed the most widespread enhanced green fluorescence protein expression and the highest number of positive nigral cells using rAAV 2/7, 2/9 and 2/1. The area transduced by rAAV 2/8 was smaller, but nevertheless almost equal numbers of nigral cells were targeted. Detailed confocal analysis revealed that serotype 2/7, 2/9, 2/1 and 2/8 transduced at least 70% of the DNs. In conclusion, these results show that various rAAV serotypes efficiently transduce nigral DNs, but significant differences in transgene expression pattern and level were observed.