Background:

Succinic 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. The SSADH-D is primarily studied using simplified model symptoms 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.

Methods:

: We utilized a CRISPR/Cas9 approach to generate human iPSC cell lines bearing ALDH5A1 mutations. Through screening, two different mutant cell lines, namely NM_001080.3: c.727_735del (p.L243_S245del) and NM_001080.3: c.730_738del (p.A244_Q246del) were obtained. Then, we induced iPSCs to neural stem cells and analyzed the characteristics of ALDH5A1 mutations in stem cells.

Results:

: The hiPSC and NSC cell lines presented typical stem cell-like morphology. We found exciting changes in ALDH5A1 expression and GABA accumulation by comparing different cell lines. In addition, we found that the mutant cell lines had a splicing variant by analyzing the cDNA between the wild-type and the mutant cell lines.

Conclusions:

: iPSC represents a promising in vitro model of SSADH-D that can be used to study early central nervous system developmental alterations and pathogenic mechanisms.