Abbott Joshua, Tambe Mitali, Pavlinov Ivan, Farkhondeh Atena, Nguyen Ha Nam, Xu Miao, Pradhan Manisha, York Tate, Might Matthew, Baumgärtel Karsten, Rodems Steven, Zheng Wei
National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, MD, United States.
Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD, United States.
Front Cell Dev Biol. 2023 Feb 16;11:1039182. doi: 10.3389/fcell.2023.1039182. eCollection 2023.
NGLY1 deficiency is an ultra-rare, autosomal recessive genetic disease caused by mutations in the gene encoding N-glycanase one that removes N-linked glycan. Patients with pathogenic mutations in NGLY1 have complex clinical symptoms including global developmental delay, motor disorder and liver dysfunction. To better understand the disease pathogenesis and the neurological symptoms of the NGLY1 deficiency we generated and characterized midbrain organoids using patient-derived iPSCs from two patients with distinct disease-causing mutations-one homozygous for p. Q208X, the other compound heterozygous for p. L318P and p. R390P and CRISPR generated NGLY1 knockout iPSCs. We demonstrate that NGLY1 deficient midbrain organoids show altered neuronal development compared to one wild type (WT) organoid. Both neuronal (TUJ1) and astrocytic glial fibrillary acid protein markers were reduced in NGLY1 patient-derived midbrain organoids along with neurotransmitter GABA. Interestingly, staining for dopaminergic neuronal marker, tyrosine hydroxylase, revealed a significant reduction in patient iPSC derived organoids. These results provide a relevant NGLY1 disease model to investigate disease mechanisms and evaluate therapeutics for treatments of NGLY1 deficiency.
NGLY1缺乏症是一种极其罕见的常染色体隐性遗传病，由编码去除N-连接聚糖的N-聚糖酶1的基因突变引起。携带NGLY1致病突变的患者具有复杂的临床症状，包括全面发育迟缓、运动障碍和肝功能障碍。为了更好地理解NGLY1缺乏症的发病机制和神经症状，我们使用来自两名患有不同致病突变的患者的诱导多能干细胞（iPSC）以及通过CRISPR技术产生的NGLY1基因敲除iPSC，构建并鉴定了中脑类器官。其中一名患者的致病突变为纯合的p.Q208X，另一名患者为p.L318P和p.R390P的复合杂合突变。我们证明，与一个野生型（WT）类器官相比，NGLY1缺乏的中脑类器官显示出神经元发育改变。在源自NGLY1患者的中脑类器官中，神经元（TUJ1）和星形胶质细胞的胶质纤维酸性蛋白标志物以及神经递质GABA均减少。有趣的是，多巴胺能神经元标志物酪氨酸羟化酶的染色显示，源自患者iPSC的类器官中该标志物显著减少。这些结果为研究NGLY1缺乏症的疾病机制和评估治疗方法提供了一个相关的疾病模型。
