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

SIELECKI AM; CAMPO VA

Congreso; XI Congreso de la Sociedad Argentina de Protozoologia; 2022

Chagas disease (or American trypanosomiasis), caused by Trypanosoma cruzi infection in humans, is aneglected disease in Latin America. The latest studies in anti-parasitic drugs have been focused in proteinacetylation. This is a reversible reaction modulated by Lysine Acetyl Transferases (KATs) and LysineDeacetylases (KDACs) enzymes. KDACs have been implicated in the modulation of the metabolicenergy pathway and gene expression. In this regard, we have previously described the effect of KDACsinhibitors and activators in T. cruzi CL Brener strain. These drugs were able to produce global changes inthe levels of acetylated histones, which showed to differentially affect the level of transcripts coding forproteins involved in differentiation and regulation of the cell cycle. Thus, we started to study the KDACsin T. cruzi (TcDACs) to determinate their role on gene expression and also it?s potential as targets forrepurposing the inhibitors or activators as anti-parasite drugs. We identified and isolated the codingsequences for the TcDACs from class I and II and quantified the transcript levels in each life stage formsin CL Brener strain. All transcripts are present in all parasite forms, with the highest level in amastigotesand epimastigotes. Also, we cloned each TcDAC coding sequence using GFP as reporter gene in theinducible system pTcINDEX, to analyze the phenotype of parasites over-expressing each enzyme indifferent strains. So far, we obtained stable transfected parasites populations for two different TcDACsclass I genes (TcCLB.511911.159 and TcCLB.504159.80 with homology to T. brucei HDAC1/KDAC1and HDAC2/KDAC2, respectively). In Dm28c, RA and Sylvio strains epimastigotes over-expressingTcKDAC1 have a perinuclear and cytoplasmic localization. We were able to over-express TcKDAC2 inSylvio strain, showing a nuclear localization. Growth curves for each transfected population showed aslight delay in replication comparing to control wild type parasites.