Chlorpromazine affects glioblastoma bioenergetics by interfering with pyruvate kinase M2

Abstract

Glioblastoma (GBM) is the most frequent and lethal brain tumor, whose therapeutic outcome - only partially effective with current schemes - places this disease among the unmet medical needs, and effective therapeutic approaches are urgently required. In our attempts to identify repositionable drugs in glioblastoma therapy, we identified the neuroleptic drug chlorpromazine (CPZ) as a very promising compound. Here we aimed to further unveil the mode of action of this drug. We performed a supervised recognition of the signal transduction pathways potentially influenced by CPZ via Reverse-Phase Protein microArrays (RPPA) and carried out an Activity-Based Protein Profiling (ABPP) followed by Mass Spectrometry (MS) analysis to possibly identify cellular factors targeted by the drug. Indeed, the glycolytic enzyme PKM2 was identified as one of the major targets of CPZ. Furthermore, using the Seahorse platform, we analyzed the bioenergetics changes induced by the drug. Consistent with the ability of CPZ to target PKM2, we detected relevant changes in GBM energy metabolism, possibly attributable to the drug’s ability to inhibit the oncogenic properties of PKM2. RPE-1 non-cancer neuroepithelial cells appeared less responsive to the drug. PKM2 silencing reduced the effects of CPZ. 3D modeling showed that CPZ interacts with PKM2 tetramer in the same region involved in binding other known activators. The effect of CPZ can be epitomized as an inhibition of the Warburg effect and thus malignancy in GBM cells, while sparing RPE-1 cells. These preclinical data enforce the rationale that allowed us to investigate the role of CPZ in GBM treatment in a recent multicenter Phase II clinical trial.

Chlorpromazine overcomes temozolomide resistance in glioblastoma by inhibiting Cx43 and essential DNA repair pathways

“…In our previous work [ 33 ], we exploited the RPPA platform to investigate the pathway-level effects of CPZ on GBM cells. Employing a similar approach, we selected key endpoints involved in DNA damage sensing (ATM, ATR) and cell cycle markers (Histone H3) and checkpoint (CHK1, PLK1, WEE1) and measured their phosphorylated forms in GBM cells either left untreated or challenged with CPZ at IC30 for 8 h. Interestingly, most CPZ-treated cell lines showed an active DNA damage response, characterized by increased phosphorylation levels of both ATM and ATR as well as CHK1 [ 50 , 52 ].…”

“…Chlorpromazine (CPZ), an FDA- and EMA-approved antipsychotic medication, has been a mainstay of clinical practice in psychiatric disorders for over seven decades, due to its well-characterized ability to antagonize dopamine at the level of the CNS dopamine receptor D2 (DRD2). In recent years, CPZ has also emerged as a promising antitumor agent, demonstrating efficacy against various in vitro-growing cancers, including GBM, where the drug is able to: (1) inhibit cell growth and proliferation; (2) induce nuclear aberrations; (3) create endoplasmic reticulum (ER) stress and ROS generation; (4) induce cytotoxic autophagy; (5) reduce stemness features; (6) interfere with GBM energy metabolism; (7) induce cancer cell death [ 29 – 33 ].…”