Dados do Trabalho

Título

EZRIN IS HIGHLY EXPRESSED AND ITS INHIBITION PRESENTS ANTINEOPLASTIC EFFECTS IN CHRONIC LYMPHOCYTIC LEUKEMIA

Introdução

Despite the development of therapeutic strategies for chronic lymphocytic leukemia (CLL), most patients remain limited. Ezrin (EZR) is a known oncogene in solid tumors, but its role in hematological malignancies is still poorly explored. There is evidence that EZR plays a key role in cell survival and activation of BCR-mediated signaling in B-cell lymphomas. In this study the role of EZR was investigated in the context of CLL.

Objetivo

In the present study, we investigated EZR expression and the potential antineoplastic effect of the EZR inhibitor NSC305787 in cellular models of CLL.

Métodos

For gene expression studies, EZR mRNA expression data from healthy donors and CLL patients were derived from the AmaZonia! PBMC CD19+ cells were collected from 56 CLL patients and healthy donors from Ribeirão Preto Medical School. For functional and molecular assays, peripheral blood samples from 24 CLL patients was obtained after informed consent and protocol approved by the Ethics Committee Nº 39510920.1.0000.5467. RNA-seq were data obtained from cBioPortal and pre-sorted using the limma-voom. Gene Set Enrichment Analysis (GSEA) performed with Hallmark gene sets. MEC-1 LLC cell line was used. NSC305787 was used for EZR pharmacological inhibition. Cell viability determined by MTT assay, apoptosis rates by annexin V/PI labeling and flow cytometry, DNA content was analyzed by PI labeling, protein expression by Western Blot. Statistical analyzes were performed by Mann-Whitney test, Student's t test or ANOVA and Bonferroni post-test. A p-value <0.05 was considered statistically significant.

Resultados

The EZR gene is highly expressed in lymphocytes derived from CLL patients (all p < 0.05) and the expression is positively associated with signaling pathways and cellular processes that contribute to the malignancy of CLL. The GSEA indicated that high EZR expression was positively associated with relevant signaling pathways associated with the development and progression of CLL, including p53, PI3K/AKT/mTOR, NFkB, and MAPK. Furthermore, we demonstrate that pharmacological inhibition of the EZR has antineoplastic effects in primary cells derived from patients with CLL. In MEC-1 cells, EZR inhibition reduced viability, clonogenicity, cell cycle progression, and induced apoptosis. EZR inhibition also reduces ERK, S6RP, and NFkB activation, indicating that EZR participates in, the activation of these signaling pathways in CLL. In conclusion, our results indicate that EZR is highly expressed in CLL and associated with molecular signatures relevant to disease development and maintenance.

Conclusões

Pharmacological inhibition of EZR attenuates several cellular and molecular behaviors associated with high risk in CLL and may present a new class of drugs to expand the treatment repertoire of this disease.

Palavras-chave

Chronic Lymphocytic leukemia, EZR, Cytoskeleton

Financiador do resumo

FAPESP, CNPq and CAPES.
2020/12909-7