Identifying early, intermediate, and late responses to GSK3a inhibition in Jurkat T-ALL cells in the presence or absence of an asparagine depletion

Amino acid availability is a crucial factor for survivability of cancer cells. Leukemia cells have been shown to resist asparagine depletion by utilizing GSK3-dependent proteasomal degradation, termed Wnt-dependent stabilization of proteins (Wnt/STOP), to replenish their amino acid pool. Inhibition of GSK3a halts the sourcing of amino acids, which subsequently leads to cancer cell vulnerability towards asparaginase therapy. Leveraging RNA sequencing we show that GSK3a inhibition leads to temporally dynamic downregulation of distinct ribosomal proteins upon amino acid starvation. To investigate the early, intermediate, and late responses to GSK3a inhibition in Jurkat T-ALL cells in the presence or absence of an asparagine depletion, a robust GSK3a knockdown was induced in Jurkat T-ALL cells using shRNA. Subsequently the cells were treated with vehicle or asparaginase, and harvested at 8, 16, 32, and 56 hours of treatment. Comparative gene expression analysis was performed compairing: timepoints, genotypes and treatment conditions.