Human iPSC-derived neural stem cells with ALDH5A1 mutation as a model of succinic semialdehyde dehydrogenase deficiency

BMC Neuroscience

Research

Xiaodan Chen¹  Minzhi Peng¹  Yanna Cai¹  Chengcheng Zhou¹  Li Liu¹ 

[1] Department of Genetics and Endocrinology, Guangzhou Women and Children’s Medical Center, Guangzhou Medical University, Guangzhou, People’s Republic of China;

关键词: SSADH-D;    Human induced pluripotent stem cells;    CRISPR/Cas9;    Neural stem cells;   

DOI  :  10.1186/s12868-022-00755-3

 received in 2022-04-20, accepted in 2022-11-11,  发布年份 2022

来源: Springer

【 摘 要 】

BackgroundSuccinic semialdehyde dehydrogenase deficiency (SSADH-D) is an autosomal recessive gamma-aminobutyric acid (GABA) metabolism disorder that can arise due to ALDH5A1 mutations, resulting in severe, progressive, untreatable neurodegeneration. SSADH-D is primarily studied using simplified models, such as HEK293 cells overexpressing genes of interest, but such overexpression can result in protein aggregation or pathway saturation that may not be representative of actual underlying disease phenotypes.MethodsWe used a CRISPR/Cas9 approach to generate human iPSC cell lines bearing ALDH5A1 mutations. Through screening, two different mutant cell lines, NM_001080.3: c.727_735del (p.L243_S245del) and NM_001080.3: c.730_738del (p.A244_Q246del), were obtained. We induced iPSCs to neural stem cells and analyzed the characteristics of ALDH5A1 mutations in stem cells.ResultsThe human iPSC and NSC cell lines presented typical stem cell–like morphology. We found changes in ALDH5A1 expression and GABA accumulation in the different cell lines. In addition, by analyzing the cDNA between the wild-type and the mutant cell lines, we found that the mutant cell lines had a splicing variant.ConclusionsiPSCs represent a promising in vitro model for SSADH-D that can be used to study early central nervous system developmental alterations and pathogenic mechanisms.

【 授权许可】

CC BY   
© The Author(s) 2022

【 图 表 】

12888_2022_4365_Article_IEq1.gif

12982_2022_119_Article_IEq90.gif

Fig. 1

12982_2022_119_Article_IEq83.gif

12982_2022_119_Article_IEq61.gif