CRISPR correction ofGBAmutation in hiPSCs restores normal function to Gaucher macrophages and increases their susceptibility toMycobacterium tuberculosis

Abstract

AbstractGaucher disease (GD) is an autosomal recessive lysosomal storage disorder caused by mutations in the β-glucocerebrosidase (GCase)GBAgene, which result in macrophage dysfunction. To investigate whether correction ofGBAmutations restores normal function to Gaucher macrophages, we performed CRISPR editing of homozygous L444P (1448T→C)GBAmutation in Type 2 GD (GBA-/-) hiPSCs, which yielded both heterozygous (GBA+/-) and homozygous (GBA+/+) isogenic lines. Macrophages derived fromGBA-/-,GBA+/- andGBA+/+ hiPSCs, were compared for GCase enzymatic activity, motility, and phagocytosis, all of which showed thatGBAmutation correction restores normal macrophage functions. Furthermore, we investigated whether lysosomal disorders drive susceptibility toMycobacterium tuberculosis, by infectingGBA-/-,GBA+/- andGBA+/+ macrophages with the virulent H37Rv lab strain. The results showed that impaired mobility and phagocytic activity of Gaucher macrophages, correlated with reduced levels of TB engulfment and TB multiplication, supporting the hypothesis that GD may be protective against tuberculosis.