Open AccessGene therapy restores dopamine transporter expression and ameliorates pathology in iPSC and mouse models of infantile parkinsonism
Author(s) -
Joanne Ng,
Serena Barral,
Carmen De La Fuente Barrigon,
Gabriele Lignani,
Fatma Aslı Erdem,
Rebecca Wallings,
Riccardo Privolizzi,
Giada Rossignoli,
Haya Alrashidi,
Sonja Heasman,
Esther Meyer,
Adeline Ngoh,
Simon Pope,
Rajvinder Karda,
Dany Perocheau,
Julien Baruteau,
Natalie Suff,
Juan Antinao Díaz,
Stephanie Schorge,
Jane Vowles,
Lucy R. Marshall,
Sally A. Cowley,
Sonja Sučić,
Michael Freissmuth,
John R. Counsell,
Richard WadeMartins,
Simon Heales,
Ahad A. Rahim,
Maximilien Bencze,
Simon Waddington,
Manju A. Kurian
Publication year - 2021
Publication title -
science translational medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 6.819
H-Index - 216
eISSN - 1946-6242
pISSN - 1946-6234
DOI - 10.1126/scitranslmed.aaw1564
Subject(s) - neuropathology , parkinsonism , dopamine transporter , dopamine , genetic enhancement , neuroscience , transporter , biology , medicine , pathology , gene , dopaminergic , disease , genetics
Most inherited neurodegenerative disorders are incurable, and often only palliative treatment is available. Precision medicine has great potential to address this unmet clinical need. We explored this paradigm in dopamine transporter deficiency syndrome (DTDS), caused by biallelic loss-of-function mutations in SLC6A3 , encoding the dopamine transporter (DAT). Patients present with early infantile hyperkinesia, severe progressive childhood parkinsonism, and raised cerebrospinal fluid dopamine metabolites. The absence of effective treatments and relentless disease course frequently leads to death in childhood. Using patient-derived induced pluripotent stem cells (iPSCs), we generated a midbrain dopaminergic (mDA) neuron model of DTDS that exhibited marked impairment of DAT activity, apoptotic neurodegeneration associated with TNFα-mediated inflammation, and dopamine toxicity. Partial restoration of DAT activity by the pharmacochaperone pifithrin-μ was mutation-specific. In contrast, lentiviral gene transfer of wild-type human SLC6A3 complementary DNA restored DAT activity and prevented neurodegeneration in all patient-derived mDA lines. To progress toward clinical translation, we used the knockout mouse model of DTDS that recapitulates human disease, exhibiting parkinsonism features, including tremor, bradykinesia, and premature death. Neonatal intracerebroventricular injection of human SLC6A3 using an adeno-associated virus (AAV) vector provided neuronal expression of human DAT, which ameliorated motor phenotype, life span, and neuronal survival in the substantia nigra and striatum, although off-target neurotoxic effects were seen at higher dosage. These were avoided with stereotactic delivery of AAV2.SLC6A3 gene therapy targeted to the midbrain of adult knockout mice, which rescued both motor phenotype and neurodegeneration, suggesting that targeted AAV gene therapy might be effective for patients with DTDS.