PIP4K2A regulates intracellular cholesterol transport through modulating PI(4,5)P2 homeostasis

The transport of LDL-derived Cholesterol from lysosomes to peroxisomes is facilitated by membrane contacts formed between the lysosomal protein synaptotagmin VII and the peroxisomal lipid phosphatidylinositol 4, 5-bisphosphate [PI(4,5)P₂]. Here, we used RNA interference to search for regulators of PI(4,5)P₂ and to study the effects of altered PI(4,5)P₂ homeostasis on Cholesterol transport. We found that knockdown of phosphatidylinositol 5-phosphate 4-kinase type-2 α (PIP4K2A) reduced peroxisomal PI(4,5)P₂ levels, decreased lysosome-peroxisome membrane contacts, and increased accumulation of lysosomal Cholesterol in human SV-589 fibroblasts. Forced expression of peroxisome-localized, kinase-active PIP4K2A in the knockdown cells reduced Cholesterol accumulation, and in vitro addition of recombinant PIP4K2A restored membrane contacts. These results suggest that PIP4K2A plays a critical role in intracellular Cholesterol transport by upregulating PI(4,5)P₂ levels in the peroxisomal membrane. Further research into PIP4K2A activity may inform future therapeutic interventions for managing lysosomal storage disorders.