Identification of H7 as a novel peroxiredoxin I inhibitor to induce differentiation of leukemia cells

Abstract

Identifying novel targets to enhance leukemia-cell differentiation is an urgent requirment. We have recently proposed that inhibiting the antioxidant enzyme peroxiredoxin I (Prdx I) may induce leukemia-cell differentiation. However, this concept remains to be confirmed. In this work, we identified H7 as a novel Prdx I inhibitor through virtual screening, in vitro activity assay, and surface plasmon resonance assay. Cellular thermal shift assay showed that H7 directly bound to Prdx I but not to Prdxs II–V in cells. H7 treatment also increased reactive oxygen species (ROS) level and cell differentiation in leukemia cells, as reflected by the upregulation of the cell surface differentiation marker CD11b/CD14 and the morphological maturation of cells. The differentiation-induction effect of H7 was further observed in some non-acute promyelocytic leukemia (APL) and primary leukemia cells apart from APL NB4 cells. Moreover, the ROS scavenger N-acetyl cysteine significantly reversed the H7-induced cell differentiation. We demonstrated as well that H7-induced cell differentiation was associated with the activation of the ROS-Erk1/2-C/EBPβ axis. Finally, we showed H7 treatment induced cell differentiation in an APL mouse model. All of these data confirmed that Prdx I was novel target for inducing leukemia-cell differentiation and that H7 was a novel lead compound for optimizing Prdx I inhibition.

CDDO-Me reveals USP7 as a novel target in ovarian cancer cells

“…The CETSA methodology is based on the thermodynamic stabilization observed for individual proteins as a result of ligand binding. Compared with other methods that confirm the interaction of small molecule compounds with proteins, CETSA is more feasible and can directly measure whether a drug molecule reaches its targets in cells [31–32, 42]. Using this method, we demonstrated that CDDO-Me interacts with USP7 in cells.…”

Isobavachalcone inhibits acute myeloid leukemia: Potential role for ROS‐dependent mitochondrial apoptosis and differentiation

“…Emerging evidence indicates that ROS as the upstream factor is closely related to apoptosis and differentiation of leukemia cells (Wei et al, 2016). Many anticancer agents induce ROS generation and trigger cancer cell apoptosis via the ROS-MAPK pathway (Simoncini et al, 2005).…”

A High-Throughput Dose-Response Cellular Thermal Shift Assay for Rapid Screening of Drug Target Engagement in Living Cells, Exemplified Using SMYD3 and IDO1

“…Confirmation of target engagement using tool inhibitors and lead optimization intermediates enabled decision making for internal drug discovery programs against diverse target classes including protein deactylase, serine/threonine/tyrosine protein kinase, lysine and arginine N-methyltransferases, oxidoreductase, dioxygenase, and E3 substrate adapter protein family members. Our enthusiasm of the broad applicability of CETSA was buoyed by numerous early reports using the technique to confirm target engagement of individual proteins comprising a wide range of target families [9][10][11][12][13][14][15][16][17][18][19][20][21][22] and the internal development of a mass spectrometry-based platform for thermal proteome profiling of the entire melting proteome. 23,24 We recognized that by replacing the low-throughput, manually intensive Western blot readout originally described with a quantitative, automated, high-density microplate format, we could provide sufficient assay capacity to fully leverage the power of CETSA and embed the technique as an integral part of hit identification and lead generation in early drug discovery campaigns.…”