EFFECTS OF DEACETYLASE INHIBITORS ON THE GROWTH OF TRITRICHOMONAS FOETUS

RIVERO, MB; LUNA, BE; SCRIMINI, S; CARRANZA, PG; ABDALA, ME; VOLTA, BJ; RIVERO, FD; LUQUE, ME; DI LULLO, D; ASSIS, MA; PRUCCA, CG

Salta

Congreso; LV Reunión Anual de la Sociedad Argentina de Investigaciones en Bioquímica y Biología Molecular; 2019

SAIB

Tritrichomonas foetus is the etiologic agent of bovine trichomoniasis, an endemic sexually transmitted disease in countries with extensive livestock with natural service as a reproductive method. Although there are few studies performed at the molecular level in this parasite, nothing is known about the existence of epigenetic regulation mechanisms. The recent sequencing of its genome has demonstrated the presence of genes homologous to enzymes that methylate, acetylate and deacetylate histones, however post-translational modifications of histones and their effects on their growth are not known. For these reasons, the aim of this work was to analyze the role of these modifying enzymes through the use of their inhibitors. Histone deacetylase enzymes can be involved in various cellular processes such as cell growth and death and can be grouped into two classes: non-NAD+- dependent (HDACs) and NAD+-dependent (Sirtuins). To evaluate their function, trichostatin A (TSA) and nicotinamide (NAM) were used, respectively. To determine the effects of HDACs inhibition by flow cytometry and immunohistochemistry, parasite cultures were incubated for 24 and 48 h at increasing concentrations of TSA (0.001 to 5uM). The IC50 results were: 19.01nM (24 h) and 11.65nM (48 h) and the LCM obtained was greater than 50nM. Once the IC50 was established, the effects at this concentration were evaluated, and the results of the cytometry analysis showed a marked change in the dispersion profile of the cells with respect to that observed in the control lots without inhibitor. This coincides with the alterations observed under the microscope: spherical cells of larger size and multinucleated. Immunohistochemistry, using the anti-acetylated tubulin α antibody, showed significant alterations at the cytoskeleton level. To assess the effects of the inhibition of sirtuins, parasite cultures were incubated for 24 and 48 h at increasing concentrations of NAM (0.1 to 25 mM). The IC50 values were: 5.43 mM (24 h) and 3.57 mM (48 h) and the LCM obtained was greater than 25 mM. The presence of homologousgenes of these enzymes and the effects observed in this study by blocking their functions, allow us to suggest for the first time that T. foetus presents this type of post-translational modifications to regulate the expression of genes that would be linked to cell growth and proliferation.