Lockdown, a selective small-molecule inhibitor of the integrin phosphatase PPM1F, blocks cancer cell invasion

Abstract

Phosphatase PPM1F is a regulator of cell adhesion by fine-tuning integrin activity and actin cytoskeleton structures. Elevated expression of this enzyme in human tumors is associated with high invasiveness, enhanced metastasis, and poor prognosis. Thus, PPM1F is a target for pharmacological intervention, yet inhibitors of this enzyme are lacking. Here, we use high-throughput screening to identify Lockdown, a reversible and non-competitive PPM1F inhibitor. Lockdown is selective for PPM1F, because this compound does not inhibit other protein phosphatases in vitro and does not induce additional phenotypes in PPM1F knockout cells. Importantly, Lockdown-treated glioblastoma cells fully re-capitulate the phenotype of PPM1F-deficient cells as assessed by increased phosphorylation of PPM1F substrates and corruption of integrin-dependent cellular processes. Ester modification yields Lockdown^Pro with increased membrane permeability and prodrug-like properties. Lockdown^Pro suppresses tissue invasion by PPM1F-overexpressing human cancer cells, validating PPM1F as a therapeutic target and providing an access point to control tumor cell dissemination.

Keywords: 6,8-difluoro-4-methyl-umbelliferyl phosphate; CAM; Ca(2+)/calmodulin-dependent kinase II; CaMKII; Cpd; DiFMUP; ECM; PAIN; PAK; PPM family; PPM1F; REOS; SAR; Ser/Thr phosphatase; WCL; chorioallantoic membrane; compound; extracellular matrix; high-throughput screen; integrin activity; pan assay interference compound; rapid elimination of swill; small-molecule inhibitor; structure activity relationship; tumor cell invasion; whole cell lysate.