Research ArticleGene Therapy

Dopamine Gene Therapy for Parkinson’s Disease in a Nonhuman Primate Without Associated Dyskinesia

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Science Translational Medicine  14 Oct 2009:
Vol. 1, Issue 2, pp. 2ra4
DOI: 10.1126/scitranslmed.3000130
  • Fig. 1

    Injection of Lenti-TH-AADC-CH1 vector to MPTP-treated macaques improves parkinsonian syndrome. Macaques treated with MPTP (black) (n = 18) were significantly impaired relative to their control pre-MPTP state, displaying severe parkinsonism as assessed by CRS (A) and VMA (B) of distance traveled. Two weeks after lentiviral injection, the animals that received Lenti-TH-AADC-CH1 (blue) (n = 6 until W8, then n = 3 until M12) demonstrated significant improvement in akinesia (B) compared to the MPTP animals that received Lenti-lacZ (red) (n = 6 until W8, then n = 3 until M12) or no viral injection (gray) (n = 6 until W8, then n = 3 until M12). W, week after gene transfer; M, month after gene transfer. *P < 0.05 relative to MPTP-Lenti-lacZ and MPTP–long term animals; **P < 0.01 relative to normal pre-MPTP lesion state. All data are expressed as the mean ± SEM.

  • Fig. 2

    Transgenes for TH, AADC, and CH1 are expressed after striatal delivery of lentiviral vectors. Immunoreactivity for TH, AADC, and CH1 was reduced by MPTP treatment, especially in the dorsal aspect of the striatum. Compare coronal brain sections from normal [left column, (A), (E), and (I)] to MPTP-Lenti-lacZ [second column, (B), (F), and (J)] macaques. Injection of the Lenti-TH-AADC-CH1 vector to MPTP-treated animals resulted in significant increases (third column) in TH (C), AADC (G), and CH1 (K) in the vicinity of the needle track in the commissural and postcommissural putamen. Higher-magnification photomicrographs of Lenti-TH-AADC-CH1–infused areas (far right column) show immunoreactive fibers throughout the putamen neuropil and neurons positive for TH (D), AADC (H), and CH1 (L). Arrows, needle tracts; white arrowhead, dorsal putamen; black arrowhead, ventral putamen; dotted line, delineation of the striatum; P, putamen; Cd, caudate nucleus. Scale bar in (A), 500 μm, applies to all panels except (D), (H), and (L); scale bar in (D), 20 μm, applies to (D), (H), and (L).

  • Fig. 3

    Daily oral l-dopa, but not Lenti-TH-AADC-CH1, induces dyskinesia in MPTP-treated animals. P < 0.01, Friedman test; *P < 0.05, post hoc Mann-Whitney test. W, week after initiation of treatment. Data are expressed as the mean ± SEM (n = 5).

  • Fig. 4

    Lenti-TH-AADC-CH1 gene therapy plus l-dopa oral treatment results in less dyskinesia than l-dopa alone in parkinsonian macaques. (A) Dyskinesia events as rated by VDA before and after l-dopa administration. Without any l-dopa, MPTP–long term (n = 3) and MPTP-Lenti-lacZ (n = 3) animals displayed spontaneous dyskinetic movements (OFF) compared to the normal (n = 3) (Mann-Whitney test, *P < 0.05) and MPTP-Lenti-TH-AADC-CH1 (n = 3) (Mann-Whitney test, **P < 0.05) animals. Acute oral l-dopa intake (20 mg/kg) significantly amplified the number of dyskinetic events (both chorea and dystonia) in MPTP–long term and MPTP-Lenti-lacZ animals (Mann-Whitney test, ***P < 0.05) compared to normal (Mann-Whitney test, **P < 0.05) and MPTP-Lenti-TH-AADC-CH1 (Mann-Whitney test, *P < 0.05) groups. (B and C) Schematic illustration of all dyskinetic events in a representative animal of each group during 120 min of observation before (B) and after (C) l-dopa administration, as rated with VDA. Each chart depicts all dystonic (upper part) and choreiform (lower part) movements in face, trunk, neck, and upper and rear limbs. Each bar represents a rated dyskinetic movement (see fig. S7).

  • Fig. 5

    Striatal injection of Lenti-TH-AADC-CH1 reduces l-dopa–induced dyskinesias in dyskinetic parkinsonian macaques. (A) Without l-dopa treatment (OFF l-dopa), animals treated with MPTP (black) (n = 6) displayed a significant -reduction in movement compared to their control pre-MPTP state. After lentiviral injections, MPTP-LID animals that received Lenti-TH-AADC-CH1 (blue) (n = 3) significantly increased their movement compared to MPTP-LID animals that received Lenti-lacZ (red) (n = 3) (Friedman test, *P < 0.05 relative to MPTP-LID-Lenti-lacZ). All data are expressed as the mean ± SEM. (B) After l-dopa treatment (ON l-dopa) (acute challenge, 20 mg/kg by mouth), MPTP-LID-Lenti-TH-AADC-CH1 (blue bars; n = 3) animals displayed a significant reduction of the total number of dyskinesias compared to MPTP-LID-Lenti-lacZ (red bars; n = 3) (Mann-Whitney test, *P < 0.05 relative to MPTP-LID-Lenti-lacZ). All data are expressed as the mean ± SEM. W, week after gene transfer; M, month after gene transfer.

  • Fig. 6

    Lenti-TH-AADC-CH1 restores striatal [DA]ec. (A) Baseline [DA]ec in normal (unlesioned, no gene transfer; white bar) and in MPTP-treated primates that received Lenti-TH-AADC-CH1 (blue bar), Lenti-lacZ (red bar), or no treatment (gray bar). Microdialysis probes were placed in the postcommissural putamen for each animal as demonstrated by in vivo T2* MRI imaging after the microdialysis procedure (see fig. S11). Lenti-TH-AADC-CH1, but not Lenti-lacZ, significantly increased striatal [DA]ec. (B) [DA]ec in the putamen at the baseline (−) and after l-dopa challenge (+) in the different groups (data are represented as AUC). (C) [l-dopa]ec in the putamen at the baseline (−) and after l-dopa challenge (+) in the different groups (data are represented as AUC). P < 0.05, Mann-Whitney test. *P < 0.05 relative to MPTP-Lenti-TH-AADC-CH1 animals; **P < 0.05, ***P < 0.01 relative to normal animals.

Additional Files

  • Supplementary Material for:

    Dopamine Gene Therapy for Parkinson's Disease in a Nonhuman Primate Without Associated Dyskinesia

    Béchir Jarraya,* Sabrina Boulet, G. Scott Ralph, Caroline Jan, Gilles Bonvento, Mimoun Azzouz, James E. Miskin, Masahiro Shin, Thierry Delzescaux, Xavier Drouot, Anne-Sophie Hérard, Denise M. Day, Emmanuel Brouillet, Susan M. Kingsman, Philippe Hantraye, Kyriacos A. Mitrophanous,* Nicholas D. Mazarakis, Stéphane Palfi*

    *To whom correspondence should be addressed. E-mail: stephane.palfi{at}; bechir.jarraya{at}; K.Mitrophanous{at}

    Published 14 October 2009, Sci. Transl. Med.2, 2ra4 (2009)
    DOI: 10.1126/scitranslmed.3000130

    This PDF file includes:

    • Experimental Design
    • Materials and Methods
    • Fig. S1. The anatomy of the basal ganglia.
    • Fig. S2. Dopamine biochemical synthesis pathway.
    • Fig. S3. Dopamine production by lentiviral vectors in HEK293T cells.
    • Fig. S4. Effect of Lenti-TH-AADC-CH1 on postural impairment in MPTP-treated macaques.
    • Fig. S5. Neuronal loss in substantia nigra pars compacta (SNpc) following systemic administration of neurotoxin MPTP.
    • Fig. S6. Dopamine transporter (DAT) immunoreactivity after striatal delivery of lentiviral vectors.
    • Fig. S7. An example of quantification of dyskinesia with video dyskinesia analysis (VDA) (The Observer 7.0 software).
    • Fig. S8. Effects of dopaminergic agents on akinesia.
    • Fig. S9. Neurotropism of EIAV lentiviral vector.
    • Fig. S10. Postmortem [DA]wt.
    • Fig. S11. In vivo localization of microdialysis probes and needle tracks.
    • Fig. S12. Stereological count of SNpc neurons after MPTP intoxication.
    • Fig. S13. Restoration of normal firing rate of basal ganglia output (globus pallidus internalis, GPi) neurons by Lenti-TH-AADC-CH1.
    • Fig. S14. Normalization of metabolic activity within the subthalamic nucleus (STN) after injection of Lenti-TH-AADC-CH1 into the motor striatum of a MPTP-treated primate.
    • Fig. S15. Astrocyte and microglia activation after striatal delivery of lentiviral vectors.
    • Videos S1-S6. Videos are available upon request and may be obtained by contacting the corresponding author directly.
    • References

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