NSC668394

NSC305787 induces a dose-dependent reduction in ALL cell viability, and is more potent than a related EZR inhibitor, NSC668394. Notably, NSC305787 shows heightened potency in ALL cells, suggesting its potential as a targeted therapy. In conclusion, our results report high EZR expression in adult ALL patients and support NSC305787 as a promising targeted therapy for ALL that should be further explored.

To study the role of RDX in chemoattraction, the inhibitor-NSC668394 suppressed collision formation and migration in BV2 cells in vitro by down-regulating F-actin. Additionally, it suppressed macrophage infiltration in infiltrating islands in vivo of intracranial tumor-bearing mice. These findings provide evidence for the role of resident cells in mediating tumor development and invasiveness and suggest that potential interacting molecules may be a strategy for controlling tumor growth by regulating the infiltration of tumor-associated microglia in the brain tumor microenvironment.

Moreover, inhibition of the CD44-Ezrin-Actin axis, using NSC668394, reduced migratory activity in hydrogels with higher HA (>0.25% w/v), while slightly increasing invasion in hydrogels with low HA (0.10 w/v%). In sum, results demonstrate the use of a matrix-mimetic, 3D hydrogel system in which to study GBM invasion through the local extracellular matrix.

Objectives Acute lymphoblastic leukemia (ALL) is a hematologic neoplasm characterized by an arrest in the process of differentiation of lymphoid precursors at an early stage and which can invade bone marrow, peripheral blood and extramedullary sites. The therapeutic options available to adult patients remain limited, and the 5-year overall survival rate is less than 45%. One of the main goals of treatment is to prevent relapses and reduce invasiveness in other organs. Recently, our research group identified the EZR gene, which encodes the ezrin protein, as an independent prognostic marker and molecular target for acute myeloid leukemia. Ezrin is an important cytoskeleton-associated protein that allows signal transduction between membrane proteins and actin filaments. The aim of the present study was to verify the impact of pharmacological inhibition of ezrin on viability, autonomous clonal growth and gene expression in ALL cell models.Material and Methods A Jurkat (mutated PTEN) NALM6 (ETV6-EBF1) and REH leukemic cell lines (ETV6-RUNX1) were treated with increasing concentrations of the ezrine inhibitors, NSC305787 and NSC668394. Cell viability was evaluated by the MTT assay, the growth of autonomous colonies by cultivation in methylcellulose in the absence of growth factors, gene expression analyzes were performed by quantitative PCR to evaluate the expression of genes related to cell cycle and apoptosis. Statistical analyzes were performed by ANOVA test and Bonferroni post-test and a p value < 0.05 was considered statistically significant.Results Treatment with both inhibitors was able to significantly reduce cell viability of Jurkat, NALM6 and REH cells in a concentration-dependent manner (p < 0.05). Compound NSC305787 had greater potency and efficacy than NSC668394, so compound NSC305787 was selected for further studies. IC50 values ​​for compound NSC305787 were 5.1 µM for Jurkat, 3.3 µM for NALM6 and 4.3 µM for REH at the 24-hour treatment time. Long-term treatment with NSC305787 reduced the autonomous clonal growth of Jurkat, NALM6 and REH cells in a concentration-dependent manner (p < 0.05). A panel of 12 genes related to cell cycle and apoptosis was investigated by quantitative RT-PCR. NSC305787 (1.6 μM for 24 hours), down-regulated CCNA2, and up-regulated CDKN1A, BCL2L1 and BIM (p < 0.05) in Jurkat cells; down-regulated CCNA2 and BCL2, and up-regulated BCL2L1, BAX and BAD in NALM6 cells; down-regulated CCNA2 and CCNB1 and up-regulated CDKN1A, CDKN1B, BCL2L1, MCL1 and BAX in REH cells.Discussion The downregulation of genes related to cell cycle progression (CCNA2 and CCNB1) and anti-apoptotic response (BCL2) and upregulation of related genes cell cycle arrest (CDKN1A, CDKN1B), pro-apoptotic genes (BAX) support the hypothesis that NSC305787 is involved in cell death induction mechanisms.Conclusion Our Results indicate that the ezrin inhibitor, NSC305787, has antileukemic effects in ALL , including reduced viability and clonogenicity. Support: CNPq, CAPES and FAPESP.

From our data we conclude that EZR expression may serve as a prognostic factor in AML. Our preclinical findings indicate that ezrin inhibitors may be employed as a putative novel class of AML targeting drugs.