Gene therapy restores dopamine transporter expression and ameliorates pathology in iPSC and mouse models of infantile parkinsonism.
Gene therapy restores dopamine transporter expression and ameliorates pathology in iPSC and mouse models of infantile parkinsonism.
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DOI：
10.1126/scitranslmed.aaw1564
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发表时间：
2021-05-19
影响因子：
17.1
通讯作者：
Kurian MA
中科院分区：
文献类型：
--
作者：
Ng J;Barral S;De La Fuente Barrigon C;Lignani G;Erdem FA;Wallings R;Privolizzi R;Rossignoli G;Alrashidi H;Heasman S;Meyer E;Ngoh A;Pope S;Karda R;Perocheau D;Baruteau J;Suff N;Antinao Diaz J;Schorge S;Vowles J;Marshall LR;Cowley SA;Sucic S;Freissmuth M;Counsell JR;Wade-Martins R;Heales SJR;Rahim AA;Bencze M;Waddington SN;Kurian MA
大多数遗传性神经退行性疾病是无法治愈的，通常只有姑息治疗可用。精准医学在解决这一未被满足的临床需求方面具有巨大潜力。我们在多巴胺转运蛋白缺乏综合征（DTDS）中探索了这种模式，该综合征是由编码多巴胺转运蛋白（DAT）的SLC6A3的双等位基因功能丧失突变引起的。患者表现为早期婴儿运动亢进，严重的进行性儿童帕金森病，脑脊液多巴胺代谢物升高。缺乏有效的治疗和无情的病程经常导致儿童死亡。利用患者来源的诱导多能干细胞（iPSCs），我们建立了DTDS的中脑多巴胺能（mDA）神经元模型，该模型显示出明显的DAT活性损伤、与tnf α介导的炎症相关的凋亡性神经变性和多巴胺毒性。药物伴侣杀虫剂-μ对DAT活性的部分恢复是突变特异性的。相比之下，野生型人SLC6A3互补DNA的慢病毒基因转移恢复了DAT活性，并防止了所有患者来源的mDA系的神经退行性变。为了在临床转化方面取得进展，我们使用了DTDS的敲除小鼠模型，该模型再现了人类疾病，表现出帕金森病的特征，包括震颤、运动迟缓和过早死亡。使用腺相关病毒（AAV）载体的新生儿脑室内注射人SLC6A3提供了人DAT的神经元表达，改善了黑质和纹状体的运动表型、寿命和神经元存活，尽管在更高剂量下观察到脱靶神经毒性作用。立体定向输送AAV2避免了这些问题。SLC6A3基因治疗靶向成年敲除小鼠的中脑，挽救了运动表型和神经变性，提示靶向AAV基因治疗可能对DTDS患者有效。
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.
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影响因子：
3.4
作者：
Illiano, Placido;Lanzo, Ambra;Di Schiavi, Elia
通讯作者：
Di Schiavi, Elia
影响因子：
5.4
作者：
Dull, T;Zufferey, R;Naldini, L
通讯作者：
Naldini, L
影响因子：
4.6
作者：
Illiano, P.;Bass, C. E.;Gainetdinov, R. R.
通讯作者：
Gainetdinov, R. R.
影响因子：
4.8
作者：
Asjad, H. M. Mazhar;Kasture, Ameya;Sucic, Sonja
通讯作者：
Sucic, Sonja
影响因子：
5.1
作者：
Charrier, S.;Ferrand, M.;Galy, A.
通讯作者：
Galy, A.