Dados do Trabalho

Título

Antineoplastic Effects of a potential hybrid inhibitor of HDAC and BTK: A combined therapeutic strategy for chronic lymphocytic leukemia (CLL)

Introdução

Chronic Lymphocytic Leukemia (CLL) is a condition characterized by an increase in the production of mature CD5+ B lymphocytes in the blood, bone marrow, spleen, and lymphoid tissues. However, these cells are dysfunctional due to genomic alterations. One of the main factors contributing to this dysfunction is the overexpression of the anti-apoptotic protein BCL2, which makes CLL cells resistant to apoptosis. To override this resistance mechanism, Venetoclax (ABT-199), a highly selective inhibitor of the BCL2 protein, has been used in clinical. Additionally, signaling through the B-cell receptor (BCR) plays a crucial role in the survival of CLL cells, along with modifications carried out by histone deacetylases (HDACs).

Objetivo

In this context, the present study aims to characterize the antineoplastic potential of a new dual hybrid inhibitor for HDACs and Bruton's tyrosine kinase (BTK) called Compound 83 (C83) in MEC-1 cell line. Furthermore, the combined use of C83 with the clinically used drug Venetoclax.

Métodos

The cell line used was MEC-1. The following assays were performed: MTT assay, clonogenic assay, annexin-V assay for apoptosis, cell cycle analysis by flow cytometry, western blot, and qPCR.

Resultados

The results showed that C83 has a lower IC50 (1.13 µM) than Vorinostat (2.1 µM), demonstrating a more significant effect on cell viability and clonal growth. It was also observed that both C83 and Vorinostat can alter the distribution of cell cycle phases in the MEC-1 cell line, depending on the concentration used. C83 demonstrated inhibitory potential in pathways involved in CLL progression. A panel of genes related to processes such as autophagy, cell cycle, mitotic checkpoint, and cell death was used to analyze the effect of C83 and Vorinostat on these cellular processes. These results were validated through protein expression analysis. It was also demonstrated that C83 increases the expression of acetylated histone H-3 and acetylated α-tubulin, indicating HDAC inhibitor activity, and decreases p-NFĸβ expression, indicating BTK inhibition. Additionally, a synergistic effect between Venetoclax and C83 was observed.

Conclusões

In summary, this study demonstrated the potential use of C83 as an inhibitor of HDACs and BTK and suggests a potential combined therapeutic strategy with Venetoclax for the treatment of CLL.

Palavras-chave

Chronic lymphocytic leukemia, HDAC inhibitor, BTK inhibitor, Venetoclax.

Financiador do resumo

FAPESP, CNPq, and CAPES.