Knockout of BTK and GCN2 followed by ASNase (asparaginase) treatment promoted apoptosis in vitro. Moreover, pharmacological inhibition of BTK by ibrutinib could also suppress GCN2 function in vitro thorough suppression of c-MYC expression. This study revealed a mechanism of synergy between the kinase inhibitor ibrutinib and ASNase treatment that could benefit ALL patients. Here, BTK is linked to acute lymphoblastic leukemia.