Data from "A new CRISPR/Cas9 GALC Knockout cellular Model Reveals Lactosylceramide Accumulation Similar to Twitcher Mice."

Krabbe disease (KD) is a lysosomal sphingolipidosis caused by the deficiency of galactocerebrosidase (GALC), leading to profound alterations in sphingolipid metabolism and severe neurodegeneration. While psychosine (Psy) accumulation is a well-established hallmark of KD, emerging evidence suggests that lactosylceramide (LacCer) may also contribute to disease pathogenesis. Here, we generated a novel human oligodendrocytic model of GALC deficiency by CRISPR/Cas9-mediated GALC knockout in MO3.13 cells and used it to investigate sphingolipid alterations associated with impaired GALC activity. GALC−/− MO3.13 cells showed a marked reduction in GALC transcript levels and enzymatic activity, confirming the successful establishment of a disease-relevant cellular model. Our results demonstrated that a) GALC knockout MO3.13 cells accumulated Psy and LacCer, and b) LacCer content was increased in the brain and heart isolated from twitcher (TWI) mice, the most widely used spontaneous animal model of KD. In parallel, AKT signalling was evaluated in both systems, supporting a link between LacCer dysregulation and altered cellular signalling pathways. Overall, these findings identify LacCer accumulation as a conserved feature of GALC deficiency across cellular and animal models and support the use of GALC−/− MO3.13 cells as a novel in vitro platform to investigate KD pathogenesis and candidate therapeutic strategies.