Mono- and Biallelic Inactivation of Huntingtin Gene in Patient-Specific Induced Pluripotent Stem Cells Reveal HTT Roles in Striatal Development and Neuronal Functions.

IF 2.1 Q3 NEUROSCIENCES Journal of Huntington's disease Pub Date : 2024-01-01 DOI:10.3233/JHD-231509
Morgane Louessard, Michel Cailleret, Margot Jarrige, Julie Bigarreau, Sophie Lenoir, Noëlle Dufour, Maria Rey, Frédéric Saudou, Nicole Deglon, Anselme L Perrier
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Abstract

Background: Mutations in the Huntingtin (HTT) gene cause Huntington's disease (HD), a neurodegenerative disorder. As a scaffold protein, HTT is involved in numerous cellular functions, but its normal and pathogenic functions during human forebrain development are poorly understood.

Objective: To investigate the developmental component of HD, with a specific emphasis on understanding the functions of wild-type and mutant HTT alleles during forebrain neuron development in individuals carrying HD mutations.

Methods: We used CRISPR/Cas9 gene-editing technology to disrupt the ATG region of the HTT gene via non-homologous end joining to produce mono- or biallelic HTT knock-out human induced pluripotent stem cell (iPSC) clones.

Results: We showed that the loss of wild-type, mutant, or both HTT isoforms does not affect the pluripotency of iPSCs or their transition into neural cells. However, we observed that HTT loss causes division impairments in forebrain neuro-epithelial cells and alters maturation of striatal projection neurons (SPNs) particularly in the acquisition of DARPP32 expression, a key functional marker of SPNs. Finally, young post-mitotic neurons derived from HTT-/- human iPSCs display cellular dysfunctions observed in adult HD neurons.

Conclusions: We described a novel collection of isogenic clones with mono- and biallelic HTT inactivation that complement existing HD-hiPSC isogenic series to explore HTT functions and test therapeutic strategies in particular HTT-lowering drugs. Characterizing neural and neuronal derivatives from human iPSCs of this collection, we show evidence that HTT loss or mutation has impacts on neuro-epithelial and striatal neurons maturation, and on basal DNA damage and BDNF axonal transport in post-mitotic neurons.

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患者特异性诱导多能干细胞中亨廷汀基因的单拷贝和双拷贝失活揭示了 HTT 在纹状体发育和神经元功能中的作用。
背景:亨廷汀(HTT)基因突变导致亨廷顿氏病(HD),这是一种神经退行性疾病。作为一种支架蛋白,HTT参与多种细胞功能,但人们对其在人类前脑发育过程中的正常功能和致病功能知之甚少:研究 HD 的发育过程,重点是了解携带 HD 基因突变的个体在前脑神经元发育过程中野生型和突变型 HTT 等位基因的功能:方法:我们使用CRISPR/Cas9基因编辑技术,通过非同源末端连接技术破坏HTT基因的ATG区域,产生单倍或双倍HTT基因敲除的人类诱导多能干细胞(iPSC)克隆:结果:我们发现,野生型、突变型或两种 HTT 同工酶的缺失不会影响 iPSC 的多能性或向神经细胞的转化。然而,我们观察到 HTT 的缺失会导致前脑神经上皮细胞分裂障碍,并改变纹状体投射神经元(SPN)的成熟,尤其是 SPN 的关键功能标记 DARPP32 的表达。最后,从HTT-/-人iPSCs衍生的年轻后有丝分裂神经元显示出成年HD神经元中观察到的细胞功能障碍:我们描述了一系列新的单拷贝和双拷贝 HTT 失活的异源克隆,它们补充了现有的 HD-hPSC 异源系列,可用于探索 HTT 功能和测试治疗策略,特别是 HTT 降解药物。通过对该系列中人类iPSC的神经和神经元衍生物进行特征描述,我们发现有证据表明,HTT缺失或突变会影响神经上皮细胞和纹状体神经元的成熟,以及有丝分裂后神经元的基础DNA损伤和BDNF轴突运输。
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来源期刊
CiteScore
4.80
自引率
9.70%
发文量
60
期刊最新文献
Changes in 24(S)-Hydroxycholesterol Are Associated with Cognitive Performance in Early Huntington's Disease: Data from the TRACK and ENROLL HD Cohorts. Somatic CAG Repeat Stability in a Transgenic Sheep Model of Huntington's Disease. Mono- and Biallelic Inactivation of Huntingtin Gene in Patient-Specific Induced Pluripotent Stem Cells Reveal HTT Roles in Striatal Development and Neuronal Functions. Alterations in Cerebrospinal Fluid Urea Occur in Late Manifest Huntington's Disease. Characterizing Heart Rate Variability Response to Maximal Exercise Testing in People with Huntington's Disease.
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