GLYCOSPHINGOLIPID BIOSINTHETIC PATHWAY AS TARGET FOR NEW ANTIPARASITIC DRUGS

T. A. PIÑERO; M. LANDONI; D.M. ACOSTA; V. G. DUSCHAK; A. S. COUTO

Potrero de los Funes

Congreso; XLVII Reunión Anual de la Sociedad Argentina de Investigación en Bioquímica y Biología Molecular; 2011

SAIB

Sphingolipids are important components of eukaryotic cells. Besides serving as structural components, they regulate cell proliferation, differentiation, cell migration, signaling, trafficking, cell-cell interactions, etc. Tamoxifen ((Z)-1-(4-(2- (dimethylamino)ethoxyl-phenyl)-1,2-diphenyl-1-butane, TAM), a nonsteroidal antiestrogen widely used for the treatment of breast cancer, has been involved with lipid metabolism in intact cells, however little experimental evidence exist up to date, related to the real effect of this drug in sphingolipid biosynthesis. Continuing with our work on the structure of Trypanosoma cruzi glycoconjugates we have shown that TAM inhibits epimastigote and trypomastigote development. We have performed a metabolic incorporation of NBD-ceramide into parasites treated or not with TAM. Sphingolipids were extracted and purified. TLC analysis showed that neutral sphingolipid biosynthesis was enhanced in treated parasites. When an HPLC analysis was performed, four labelled components were observed and interestingly three of them of the treated parasites presented a double area when compared with the untreated ones. Each component was isolated and further analyzed by UV-MALDI-TOF mass spectrometry. In addition, as it is well known that ceramide induces cell apoptosis, we further analyzed by flow cytometry if this event is taking place in T. cruzi.Trypanosoma cruzi glycoconjugates we have shown that TAM inhibits epimastigote and trypomastigote development. We have performed a metabolic incorporation of NBD-ceramide into parasites treated or not with TAM. Sphingolipids were extracted and purified. TLC analysis showed that neutral sphingolipid biosynthesis was enhanced in treated parasites. When an HPLC analysis was performed, four labelled components were observed and interestingly three of them of the treated parasites presented a double area when compared with the untreated ones. Each component was isolated and further analyzed by UV-MALDI-TOF mass spectrometry. In addition, as it is well known that ceramide induces cell apoptosis, we further analyzed by flow cytometry if this event is taking place in T. cruzi.T. cruzi.