This technology includes a high-yield, easy-to-culture mouse neuronal cell model with nearly complete glucocerebrosidase deficiency representative of Gaucher disease (GD) to study pathophysiology and evaluate new therapies. GD is an autosomal recessive lysosomal storage disorder caused by loss-of function mutations in the GBA1 gene, which codes for the lysosomal hydrolase glucocerebrosidase (GCase). Inventors have successfully immortalized cortical neurons from embryonic null allele GBA-/- mice and the control littermate (GBA+/+) by infecting differentiated primary cortical neurons in culture with an EF1aSV40T lentivirus. Immortalized GBA-/- neurons lack glucocerebrosidase protein and enzyme activity and exhibit a dramatic increase in glucosylceramide and glucosylsphingosine accumulation,enlarged lysosomes, and an impaired ATP-dependent calcium-influx response.
The null allele GBA-/- mouse neuronal cell line model provides a much-needed tool to study the pathophysiology of Gaucher disease and to evaluate new therapies. Because mutations in GBA1 are also a genetic risk factor for Parkinson disease (PD), dementia with Lewy bodies (DLB), and multiple system atrophy (MSA), this cell line may also provide a basis to explore the molecular mechanism of these disorders and other synucleinopathies.
This new cell line exhibits a dramatic increase in glucosylceramide and glucosylsphingosine accumulation, enlarged lysosomes, and an impaired ATP-dependent calcium-influx response, compared to other cell lines which have limitations such as low yield, difficulty in propagation, and introduction of exogenous mutant GBA 1.