INVESTIGADORES
CARRILLO carolina
congresos y reuniones científicas
Título:
Anthracylines effects on polyamine uptake and proliferation on T. cruzi epimastigotes
Autor/es:
RUIZ MD; FRACCAROLI L; BALCAZAR DE; SBARAGLINI ML; LAROCCA L; CARRILLO C
Lugar:
CABA
Reunión:
Congreso; Drug Discovery for Neglected Diseases International Congress 2018 4th Scientific Meeting of the Research Network Natural Products against Neglected Diseases; 2018
Institución organizadora:
DDNDIC
Resumen:
Chagas disease is an endemic parasitosis originally from Latin America, caused by the protozoan Trypanosoma cruzi(T. cruzi). The current therapies are limited in efficacy and show multiple side effects. Thus, there is a need to identifynew effective and specific trypanocidal strategies.Anthracyclines, used as anticancer agents, exert their cytotoxicity by blocking DNA metabolism. In humans, these aretransported into cells by the organic cation transporter hOCT1 [1]. Previous results have shown that these compoundsinhibit DNA topoisomerases from trypanosomatids [2], however no specific transporters have been identified.As hOCT1 functions as a low affinity polyamine permease, we were interested in analyzing the interaction and/orincorporation of anthracyclines in T. cruzi by the putrescine permease TcPAT12 [3], a member of the TcAAAP transportersfamily [4].Anthracyclines tested were Daunorubicin (Dnr) and Doxorubicin (Dxr). We evaluated T. cruzi epimastigotes proliferationand viability by growth curves and MTT assay under different culture conditions. The strains assayed were Y-GFP(control) and Y-TcPAT12-GFP (overexpressing TcPAT12 permease).To evaluate the effects of Dnr y Dxr, transport assays were performed for putrescine (TcPAT12), arginine (TcAAAP411)and riboflavin (TcRibj) [5].Dnr and Dxr significantly decreased T. cruzi epimastigotes proliferation rate, in a dose dependent manner, assesedby MTT and cell counting using a Neubauer chamber. IC50 values ranged between 0.3 µM for Dnr and 3 µM for Dxrin control and Y-TcPAT12-GFP strains. These strains became more sensitive to Dnr and Dxr when epimastigotes wereputrescine deprived during 15 days (IC50 values ranging 0.1 µM for Dnr and 2 µM for Dxr).Putrescine uptake was affected by Dnr and Dxr in both strains under study. Percentage of putrescine uptake respectto untreated epimastigotes: control + Dnr 50µM: 46.4±8.8% vs TcPAT12 + Dnr 50µM: 78.2±2.5%; control + Dxr 50µM:59.9±2.4% vs TcPAT12 + Dxr 50µM: 91.4±8.6%. Arginine and riboflavin uptake were not affected under these conditions.The findings presented herein showed that Dnr and Dxr affect T. cruzi epimastigotes survival and proliferation. This effectcould be related to the decrease of polyamine uptake and/or to anthracyclines incorporation by TcPAT12 transporterand their intracellular toxic effects.References:1. Andreev E., et al. 2016. Sci Rep, 6:205082. Das A., et al. 2004. Trends Parasitol, 20(8):381-7