背景:不同的神经亚型在多种神经退行性疾病中选择性丧失。亨廷顿氏病(HD)是一种遗传性神经退行性疾病,其特征是主要影响纹状体的运动异常。亨廷顿(HTT)突变涉及扩展的CAG重复,导致不溶性聚氧乙烯醚,这使得GABA+中等多刺神经元(MSN)更容易受到细胞死亡的影响。人类多能干细胞(hPSC)技术允许构建疾病特异性模型,为研究发病机制提供有价值的细胞模型,药物筛选,和高通量分析。
方法:在本研究中,我们建立了一种方法,可以在21天内从hPSC衍生的神经祖细胞中快速有效地产生MSNs(>90%),通过引入转录因子的特定组合。
结果:我们有效地诱导了几种神经亚型,并行,基于相同的细胞来源,并透露,与其他神经亚型相比,MSN表现出更高的polyQ聚集倾向和过表达毒性,BDNF/TrkB信号传导更严重的功能障碍,对BDNF戒断的敏感性更大,和更严重的核质运输(NCT)紊乱。我们进一步发现,核层蛋白LMNB1在HD神经元中大大减少,并错定位于细胞质和轴突。敲除HTT或用KPT335治疗,KPT335是一种口服选择性核出口抑制剂(SINE),有效地减弱病理表型,减轻BDNF戒断引起的神经元死亡。
结论:因此,本研究建立了获得MSN的有效方法,并强调了使用高纯度MSN研究HD发病机制的必要性,特别是MSN选择性漏洞。
BACKGROUND: Different neural subtypes are selectively lost in diverse neurodegenerative diseases. Huntington\'s disease (HD) is an inherited neurodegenerative disease characterized by motor abnormalities that primarily affect the striatum. The Huntingtin (HTT) mutation involves an expanded CAG repeat, leading to insoluble polyQ, which renders GABA+ medium spiny neurons (MSN) more venerable to cell death. Human pluripotent stem cells (hPSCs) technology allows for the construction of disease-specific models, providing valuable cellular models for studying pathogenesis, drug screening, and high-throughput analysis.
METHODS: In this study, we established a method that allows for rapid and efficient generation of MSNs (> 90%) within 21 days from hPSC-derived neural progenitor cells, by introducing a specific combination of transcription factors.
RESULTS: We efficiently induced several neural subtypes, in parallel, based on the same cell source, and revealed that, compared to other neural subtypes, MSNs exhibited higher polyQ aggregation propensity and overexpression toxicity, more severe dysfunction in BDNF/TrkB signaling, greater susceptibility to BDNF withdrawal, and more severe disturbances in nucleocytoplasmic transport (NCT). We further found that the nuclear lamina protein LMNB1 was greatly reduced in HD neurons and mislocalized to the cytoplasm and axons. Knockdown of HTT or treatment with KPT335, an orally selective inhibitor of nuclear export (SINE), effectively attenuated the pathological phenotypes and alleviated neuronal death caused by BDNF withdrawal.
CONCLUSIONS: This study thus establishes an effective method for obtaining MSNs and underscores the necessity of using high-purity MSNs to study HD pathogenesis, especially the MSN-selective vulnerability.