The PIP4K2 inhibitor THZ-P1-2 exhibits antileukemia activity by disruption of mitochondrial homeostasis and autophagy

Abstract

The treatment of acute leukemia is challenging because of the genetic heterogeneity between and within patients. Leukemic stem cells (LSCs) are relatively drug-resistant and frequently relapse. Their plasticity and capacity to adapt to extracellular stress, in which mitochondrial metabolism and autophagy play important roles, further complicates treatment. Genetic models of phosphatidylinositol-5-phosphate 4-kinase type 2 protein (PIP4K2s) inhibition have demonstrated the relevance of these enzymes in mitochondrial homeostasis and autophagic flux. Here, we uncovered the cellular and molecular effects of THZ-P1-2, a pan-inhibitor of PIP4K2s, in acute leukemia cells. THZ-P1-2 reduced cell viability and induced DNA damage, apoptosis, loss of mitochondrial membrane potential, and the accumulation of acidic vesicular organelles. Protein expression analysis revealed that THZ-P1-2 impaired autophagic flux. In addition, THZ-P1-2 induced cell differentiation and showed synergistic effects with venetoclax. In primary leukemia cells, LC-MS/MS-based proteome analysis revealed that sensitivity to THZ-P1-2 is associated with mitochondrial metabolism, cell cycle, cell-of-origin (hematopoietic stem cell and myeloid progenitor), and the TP53 pathway. The minimal effects of THZ-P1-2 observed in healthy CD34+ cells suggest a favorable therapeutic window. Our study provides insights into the pharmacological inhibition of PIP4K2s targeting mitochondrial homeostasis and autophagy, shedding light on a new class of drugs for acute leukemia.

PIP4K2C inhibition reverses autophagic flux impairment induced by SARS-CoV-2

“…3e). THZ-P1-2 and CVM-05-002, inhibitors of PIKfyve and PIP4K2C 33, 34 , also dose-dependently inhibited SARS-CoV-2 infection (EC 50 =0.2 µM and EC 50 =0.97 µM, respectively), albeit THZ-P1-2 demonstrated greater cytotoxicity (CC 50 =16 µM) (Fig. 3e).…”

Identification of ARUK2002821 as an isoform-selective PI5P4Kα inhibitor

“…Various PI5P4K inhibitors have been reported to have a therapeutic effect in oncology settings. 18,19 Covalent pan-PI5P4K inhibitor THZ-P1-2 ( 1 ) was shown to have anti-proliferative activity in acute myeloid and lymphoblastic leukaemias (AML/ALL) 19–22 and the small molecule A131 ( 2 ), which was shown to target mitotic pathways and PI5P4Ks, was effective in cancer cell-specific lethality 18 (Fig. 1).…”

The rational design of ARUK2007145, a dual inhibitor of the α and γ isoforms of the lipid kinase phosphatidylinositol 5-phosphate 4-kinase (PI5P4K)

“…Several recent reports detail inhibitors for PI5P4Ks, not only pan-specific 25–29 but also isoform specific. 30–34 Emerging tools are also being developed for dual-specific inhibitors for PI5P4Kα and PI5P4Kβ 22,35 and methods have also been reported for removing protein completely in cells using PROTAC systems, such as JWZ-1-80 for PI5P4Kγ, 36 which will be useful to interrogate non-catalytic roles.…”