Multicistronic Lentiviral Vector-Mediated Striatal Gene Transfer of Aromatic l-Amino Acid Decarboxylase, Tyrosine Hydroxylase, and GTP Cyclohydrolase I Induces Sustained Transgene Expression, Dopamine Production, and Functional Improvement in a Rat Model of Parkinson's Disease.

Abstract

Parkinson's disease (PD) is a neurodegenerative disorder characterized by the selective loss of dopaminergic neurons in the substantia nigra. This loss leads to complete dopamine depletion in the striatum and severe motor impairment. It has been demonstrated previously that a lentiviral vector system based on equine infectious anemia virus (EIAV) gives rise to highly efficient and sustained transduction of neurons in the rat brain. Therefore, a dopamine replacement strategy using EIAV has been investigated as a treatment in the 6-hydroxydopamine (6-OHDA) animal model of PD. A self-inactivating EIAV minimal lentiviral vector that expresses tyrosine hydroxylase (TH), aromatic amino acid dopa decarboxylase (AADC), and GTP cyclohydrolase 1 (CH1) in a single transcription unit has been generated. In cultured striatal neurons transduced with this vector, TH, AADC, and CH1 proteins can all be detected. After stereotactic delivery into the dopamine-denervated striatum of the 6-OHDA-lesioned rat, sustained expression of each enzyme and effective production of catecholamines were detected, resulting in significant reduction of apomorphine-induced motor asymmetry compared with control animals (p < 0.003). Expression of each enzyme in the striatum was observed for up to 5 months after injection. These data indicate that the delivery of three catecholaminergic synthetic enzymes by a single lentiviral vector can achieve functional improvement and thus open the potential for the use of this vector for gene therapy of late-stage PD patients.

Gene therapy progress and prospects: Parkinson's disease

“…Nonviral gene transfer (liposomes or naked plasmids) is in general ineffective for gene transfer to the brain parenchyma, but each of the major viral vectors have demonstrated utility in experimental models of PD. In both earlier and recent studies viral vectors based on Ad1, 42 lentivirus (LV), 11,20 AAV 13,19 and HSV 15,24 have all been used to transfer relevant genes to the substantia nigra or striatum of experimental animals. Owing to the differences between studies in the animal model of PD employed, the site and volume of vector inoculation, the transgene and promoter constructs tested, the vector dose and the outcome measures assessed, the published literature does not allow one to make a direct comparison between vectors.…”

Development of inducible EIAV-based lentiviral vector packaging and producer cell lines

“…[1][2][3][4][5] EIAV based LVs are being developed for a number of different gene therapy applications for a variety of diseases. [6][7][8][9][10][11][12][13] Recent focus has been on the development of ProSavin, which is an EIAV-based LV for the treatment of Parkinson's disease encoding three enzymes required for the dopamine synthesis. 6 Currently, ProSavin is in phase I/II clinical trials that require relatively small amounts of vector material.…”

“…[6][7][8][9][10][11][12][13] Recent focus has been on the development of ProSavin, which is an EIAV-based LV for the treatment of Parkinson's disease encoding three enzymes required for the dopamine synthesis. 6 Currently, ProSavin is in phase I/II clinical trials that require relatively small amounts of vector material. This was generated by transient co-transfection of human embryonic kidney 293T (HEK293T) cells with three plasmids that encode the necessary components for vector production (Vector genome, EIAV Gag/Pol and a heterologous envelope (VSV-G)).…”

Viral vectors for in vivo gene transfer in Parkinson's disease: Properties and clinical grade production

“…Recombinant adenovirus (rAd), recombinant adenoassociated viruses (rAAV), herpes simplex viruses (HSV) and lentiviruses (Lv) are currently most utilized viral vectors that have been demonstrated to reliably transfer DNA in animal models of PD (Azzouz et al, 2002;Bilang-Bleuel et al, 1997;Choi-Lundberg et al, 1997;Corti et al, 1999;Kordower et al, 2000). Currently, there are at least 3 early stage clinical trials testing the safety of vector-delivery of genes to treat PD (http://www.gemcris.od.nih.gov/).…”