DNAJC6 encodes auxilin, a co-chaperone protein involved in clathrin-mediated endocytosis (CME) at the presynaptic terminal. Biallelic mutations in DNAJC6 cause a complex, early-onset neurodegenerative disorder characterized by rapidly progressive parkinsonism-dystonia in childhood. The disease is commonly associated with additional neurodevelopmental, neurological and neuropsychiatric features. Currently, there are no disease-modifying treatments for this condition, resulting in significant morbidity and risk of premature mortality. To investigate the underlying disease mechanisms in childhood-onset DNAJC6 parkinsonism, we generated induced pluripotent stem cells (iPSC) from three patients harboring pathogenic loss-of-function DNAJC6 mutations and subsequently developed a midbrain dopaminergic (mDA) neuronal model of disease. When compared to age-matched and CRISPR-corrected isogenic controls, the neuronal cell model revealed disease-specific auxilin deficiency as well as disturbance of synaptic vesicle (SV) recycling and homeostasis. We also observed neurodevelopmental dysregulation affecting ventral midbrain patterning and neuronal maturation. In order to explore the feasibility of a viral vector-mediated gene therapy approach, iPSC-derived neuronal cultures were treated with lentiviral DNAJC6 gene transfer, which restored auxilin expression and rescued CME. Our patient-derived neuronal model provides deeper insights into the molecular mechanisms of auxilin deficiency as well as a robust platform for the development of targeted precision therapy approaches.

中文翻译：

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DNAJC6 帕金森病的神经发育和突触缺陷，适合基因治疗

DNAJC6 编码辅助蛋白，这是一种参与突触前末端网格蛋白介导的内吞作用 (CME) 的辅助伴侣蛋白。DNAJC6 的双等位基因突变会导致一种复杂的早发性神经退行性疾病，其特征是儿童时期快速进展的帕金森病-肌张力障碍。该疾病通常与额外的神经发育、神经学和神经精神特征相关。目前，还没有针对这种情况的疾病缓解治疗方法，导致显着的发病率和过早死亡的风险。为了研究儿童期发病的 DNAJC6 帕金森病的潜在疾病机制，我们从三名携带致病性功能丧失 DNAJC6 突变的患者中产生了诱导多能干细胞 (iPSC)，随后开发了中脑多巴胺能 (mDA) 神经元疾病模型。与年龄匹配和 CRISPR 校正的同基因对照相比，神经元细胞模型揭示了疾病特异性辅助素缺乏以及突触小泡 (SV) 循环和稳态的紊乱。我们还观察到影响腹侧中脑模式和神经元成熟的神经发育失调。为了探索病毒载体介导的基因治疗方法的可行性，用慢病毒 DNAJC6 基因转移处理 iPSC 衍生的神经元培养物，这恢复了辅助素表达并挽救了 CME。我们的源自患者的神经元模型为辅助素缺乏的分子机制提供了更深入的见解，并为开发靶向精准治疗方法提供了强大的平台。