Gene therapy for Childhood Parkinsonism: Dopamine transporter deficiency syndrome

儿童帕金森症的基因治疗：多巴胺转运蛋白缺乏综合征

• 批准号: MR/R015325/1
• 负责人: Simon Waddington
• 金额: $ 63.1万
• 依托单位: University College London
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2018
• 资助国家: 英国
• 起止时间: 2018 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=MR%2FR015325%2F1
• 关键词: Gene; therapy; Childhood; Parkinsonism; Dopamine

Dopamine transporter deficiency syndrome (DTDS) is a devastating childhood neurotransmitter disorder. It clinically presents with progressive infantile parkinsonism-dystonia and is frequently misdiagnosed as cerebral palsy. The children are severely disabled with inability to control movement experiencing distressing hyperkinesia and dystonic postures. They progressively worsen with severe slowing of movements and muscle rigidity, termed parkinsonism. This results in severe neurodisability and is untreatable with medication or surgery, with a third dying by mid-adolescence. DTDS results from loss of function mutations the SLC6A3 gene coding for the Dopamine transporter (DAT). DAT functions to uptake dopamine to terminate dopamine neurotransmission and uptake dopamine for recycling and is physiologically expressed in specific brain regions (midbrain and striatum).There is medical need to develop impactful treatment for this untreatable childhood neurological disorders. We aim to develop a potential cure for DTDS by delivering Adeno associated virus (AAV) mediated hDAT gene therapy. This will result in expression of normally functioning DAT to restore dopamine homeostasis to improve motor function, health and neurodisability of DTDS children. Gene therapy for childhood neurological disorders is showing great promise. Adeno associated virus serotype 9 (AAV9) delivered intravenously rescued the Spinal muscular atrophy (SMA) mouse in 2010 and progressed to clinical trial within 4 years (NCT02122952). SMA type 1 is severe motor neuron disorder whereby affected babies do not achieve sitting and most die by 2years. Fifteen treated children have shown striking motor improvements, with 12 able to sit and some walking independently. Gene therapy is used to treat another childhood neurotransmitter disorder called Aromatic L-amino acid decarboxylase (AADC) deficiency, that is clinically similar to DTDS. Initially 4 children were treated by direct (stereotactic) injection of AAV2-AADC to a specific dopaminergic brain region called the putamen. This established clinical safety and all 4 children showed motor improvement. A subsequent phase II clinical trial has treated 18 children to date. Furthermore over 100 patients with Parkinson's disease, have safely received AAV2 gene therapy vectors delivered by stereotactic injection to the striatum or midbrain over the last decade. AAV2 gene therapies for Parkinson's disease have delivered dopamine synthesis genes to increase dopamine synthesis and provide good clinical evidence for the safety of AAV2 vectors and stereotactic injection. We previously undertook proof of principle gene therapy study for DTDS using the DAT-KO mouse model. This DAT-KO mouse recapitulates DTDS symptoms with early hyperlocomotion, 59% entering a Parkinsonian phase with bradykinesia, tremor, weight loss and reduced survival of 41%, by 5 weeks old. We delivered AAV9 gene therapy to neonatal DAT-KO mice by injection to the intracerebroventrical. This rescued the DAT-KO motor phenotype and survival. However with this approach, widespread intracranial expression of DAT was achieved, with significant off-target expression. We identified that off-target expression was associated with neuronal loss and reactive astrogliosis in the cortex. We therefore concluded that for clinical application, it is paramount to restrict hDAT expression to dopaminergic neurons. We aim to improve safety levels by changing AAV serotype to AAV2 that has a restricted expression profile compared to AAV9. We will also inject the vector into the dopaminergic brain regions affected in DTDS (midbrain and striatum). This is a logical step towards clinical translation whereby AAV2 and stereotactic injection methods have established clinical safety. We will evaluate expression, efficacy, and safety of stereotactically delivered AAV2.hDAT in adult DAT-KO mice as proof of concept preclinical development of gene therapy for DTDS.

多巴胺转运蛋白缺乏综合征（DTDS）是毁灭性的儿童神经递质疾病。它在临床上表现出了进步的婴儿帕金森氏症 - dystonia，并且经常被误诊为脑瘫。这些孩子严重残疾，无法控制运动的运动和肌张力肌的痛苦。他们随着运动和肌肉僵硬的严重减缓而逐渐恶化，称为帕金森主义。这会导致严重的神经疾病性，并且在药物或手术中无法治疗，第三次死亡到青春期。 DTDS是由于功能突变的丧失导致多巴胺转运蛋白（DAT）的SLC6A3基因编码。 DAT功能可用于摄取多巴胺以终止多巴胺神经传递和摄取多巴胺进行回收，并在特定的大脑区域（中脑和纹状体）在生理上表达。医学上需要对这种不可培养的儿童期神经疾病产生影响的治疗方法。我们旨在通过输送Adeno相关病毒（AAV）介导的HDAT基因治疗来开发DTD的潜在治疗。这将导致表达正常运转的DAT，以恢复多巴胺稳态，以改善DTDS儿童的运动功能，健康和神经疾病。儿童神经系统疾病的基因疗法表现出巨大的希望。 Adeno相关的病毒血清型9（AAV9）在2010年静脉内提供了脊柱肌肉萎缩（SMA）小鼠，并在4年内进行了临床试验（NCT02122952）。 SMA 1型是严重的运动神经元疾病，因此受影响的婴儿无法坐着，大多数人死于2年。 15个经过治疗的儿童表现出惊人的运动改善，有12个能够坐下，有些可以独立行走。基因治疗用于治疗一种称为芳香族L-氨基酸脱羧酶（AADC）缺乏症的儿童神经递质疾病，该疾病在临床上与DTD相似。最初，有4名儿童通过直接（立体定向）将AAV2-AADC注射到一个称为pepamen的特定多巴胺能大脑区域。这已经建立的临床安全，所有4个儿童均表现出运动的改善。随后的II期临床试验已经治疗了18名儿童。此外，在过去的十年中，有100多名帕金森氏病患者安全地接受了立体定向注射给纹状体或中脑的AAV2基因治疗载体。帕金森氏病的AAV2基因疗法已递送多巴胺合成基因，以增加多巴胺合成，并为AAV2载体和立体定向注射的安全提供良好的临床证据。我们先前使用Dat-KO小鼠模型进行了DTD的原理基因治疗研究证明。这款DAT-KO小鼠以早期超稳态概括了DTDS症状，59％的人进入帕金森氏症阶段，患有Bradykinesia，震颤，体重减轻，生存率降低了41％，到5周大。我们通过注射到脑内桥术中将AAV9基因治疗传递给新生儿DAT-KO小鼠。这拯救了Dat-Ko运动表型和生存。然而，通过这种方法，达到了DAT的广泛颅内表达，并具有明显的脱靶表达。我们确定靶向表达与皮质中的神经元丧失和反应性星形胶质症有关。因此，我们得出的结论是，对于临床应用，将HDAT表达限制为多巴胺能神经元至关重要。我们旨在通过将AAV血清型更改为与AAV9相比具有限制的表达曲线的AAV血清型来提高安全水平。我们还将将载体注入DTD（中脑和纹状体）影响的多巴胺能脑区域。这是朝着临床翻译迈出的逻辑步骤，在该临床翻译中，AAV2和立体定向注射方法已经确立了临床安全性。我们将评估成年Dat-Ko小鼠中立体定位递送AAV2.HDAT的表达，功效和安全性，作为DTD基因治疗临床前开发的概念证明。

项目成果

High-efficiency transduction of spinal cord motor neurons by intrauterine delivery of integration-deficient lentiviral vectors.

• DOI: 10.1016/j.jconrel.2017.12.029
• 发表时间: 2018-03-10
• 期刊: Journal of controlled release : official journal of the Controlled Release Society
• 作者: Ahmed SG;Waddington SN;Boza-Morán MG;Yáñez-Muñoz RJ
• 通讯作者: Yáñez-Muñoz RJ

Argininosuccinic aciduria fosters neuronal nitrosative stress reversed by Asl gene transfer

精氨基琥珀酸尿症促进 Asl 基因转移逆转神经元亚硝化应激

• DOI: 10.1101/348292
• 发表时间: 2018
• 作者: Baruteau J

Enhancement of mouse hematopoietic stem/progenitor cell function via transient gene delivery using integration-deficient lentiviral vectors.

• DOI: 10.1016/j.exphem.2017.09.003
• 发表时间: 2018-01
• 期刊: Experimental hematology
• 影响因子: 2.6
• 作者: Alonso-Ferrero ME;van Til NP;Bartolovic K;Mata MF;Wagemaker G;Moulding D;Williams DA;Kinnon C;Waddington SN;Milsom MD;Howe SJ
• 通讯作者: Howe SJ

Urea Cycle Related Amino Acids Measured in Dried Bloodspots Enable Long-Term In Vivo Monitoring and Therapeutic Adjustment.

在干血斑中测量尿素循环相关氨基酸可以实现长期体内监测和治疗调整。

• DOI: 10.3390/metabo9110275
• 发表时间: 2019
• 期刊: Metabolites
• 影响因子: 4.1
• 作者: Baruteau J

Non-invasive somatotransgenic bioimaging in living animals

活体动物的非侵入性体细胞转基因生物成像

• DOI: 10.12688/f1000research.25274.1
• 发表时间: 2020
• 期刊: F1000Research
• 作者: Delhove J