Last modified: 2018-09-09
Abstract
Purpose
Vanishing white matter disease (VWM) is one of the most prevalent inherited leukoencephalopathies in children. Although the defects are in the housekeeping genes EIF2B1–5, VWM is primarily a leukoencephalopathy. No model for VWM induced pluripotent cells (iPSCs) has been established for exploring the mechanism of VWM.
Materials and Methods
2 VWM iPSCs model was established by a virus-free nonintegrating episomal system. Wild-type and VWM iPSCs were sequentially differentiated into neural stem cells (NSCs), then into neurons, oligodendrocytes and astrocytes. Apoptosis was detected by Annexin V/PI.
Results
(1) Two VWM iPSCs models were successfully established and identified: alkaline phosphatase staining was positive and iPSCs expressed pluripotent markers NANOG and SSEA4. Moreover, teratoma assay showed that iPSCs have potential of differentiation into three germ layers. (2) Wild-type and VWM iPSCs derived NSCs can differentiated into neurons and oligodendrocytes. (3) VWM iPSCs derived astrocytes revealed dysfunction. 1) VWM iPSC-derived astrocytes were dysmorphic, manifested as shorter processes. 2) Total GFAP and α GFAP of VWM astrocytes were fewer than wild-type in mRNA level, whereas δ GFAP was much higher. 3) The early and total apoptosis rates of wild-type, VWM1, and VWM2 astrocytes were higher than those of the wild-type astrocytes, p<0.05.
Conclusion
The establishment of VWM-specific iPSC models provides a platform for exploring the pathogenesis of VWM and future drug screening. The results suggested that in vitro differentiation of wild-type and VWM iPSCs into NSCs, neurons, and OLs showed no significant difference. Whereas, VWM astrocytes exhibited abnormal morphology and dysfunction.