Characterization of Trypanosoma cruzi lysine deacetylase enzymes and evaluation as potential targets for anti-parasitic drugs.

ALEJANDRO MARTIN SIELECKI; VANINA A. CAMPO

Jornada; Primera Jornada Itinerante de Epigenética; 2022

Chagas disease (or American trypanosomiasis), caused by Trypanosoma cruzi infection in humans, is a neglected disease in Latin America. The latest studies in anti-parasitic drugs have been focused in protein acetylation. This is a reversible reaction modulated by Lysine Acetyl Transferases (KATs) and Lysine Deacetylases (KDACs) enzymes. KDACs have been implicated in the modulation of the metabolic energy pathway and gene expression. In this regard, we have previously described the effect of KDACs inhibitors and activators in T. cruzi CL Brener strain. These drugs were able to produce global changes in the levels of acetylated histones, which showed to differentially affect the level of transcripts coding for proteins involved in differentiation and regulation of the cell cycle. Thus, we started to study the KDACs in T. cruzi (TcDACs) to determinate their role on gene expression and also it’s potential as targets for repurposing the inhibitors or activators as anti-parasite drugs. We identified and isolated the coding sequences for the TcDACs from class I and II and quantified the transcript levels in each life stage forms in CL Brener strain. All transcripts are present in all parasite forms, with the highest level in amastigotes and epimastigotes. Also, we cloned each TcDAC coding sequence using GFP as reporter gene in the inducible system pTcINDEX, to analyze the phenotype of parasites over-expressing each enzyme in different strains. So far, we obtained stable transfected parasites populations for two different TcDACs class I genes (TcCLB.511911.159 and TcCLB.504159.80 with homology to T. brucei HDAC1/KDAC1 and HDAC2/KDAC2, respectively). In Dm28c, RA and Sylvio strains epimastigotes over-expressing TcKDAC1 have a perinuclear and cytoplasmic localization. We were able to over-express TcKDAC2 in Sylvio strain, showing a nuclear localization. Growth curves for each transfected population showed a slight delay in replication comparing to control wild type parasites.