ELIGLUSTAT INHIBITS GLUCOSYLCERAMIDE SYNTHASE AND GLOBOTRIAOSYLCERAMIDE EXPRESSION WITHOUT INTERACTING WITH SHIGA TOXIN 2

SANCHEZ DAIANA; JUAN JOSÉ CASAL; CRISTINA IBARRA; MARIA MARTA AMARAL; CLAUDIA SILBERSTEIN

Buenos Aires

Congreso; Reunión de Sociedades de Biociencias; 2021

Sociedad Argentina de Investigación Clínica

Shiga toxin-producing Escherichia coli is responsible for HemolyticUremic Syndrome (HUS), a cause of renal failure in children. Renaldamage has been associated with Shiga toxin (Stx), which binds tothe globotriaosylceramide (Gb3) receptor on the plasma membraneof target cells. We have previously shown that Eliglustat (EG), aninhibitor of glucosylceramide synthase LS , the rst step of gly-cosphingolipids pathway, also inhibits Gb3 expression and preventsthe cytotoxic effects of Shiga toxin type 2 (Stx2) in human corticalrenal tubular epithelial cells RTEC primary cultures. The aim ofthis work was to clarify the mode of interaction of EG in the ac-tive site of GLS and its possible interaction with Stx2 and Gb3, andcompare to b3 expression in RTEC treated with E and Stx2.For this, a computational procedure called molecular docking wascarried out with Smina software and Gibbs free energy was calculat-ed for determining the stability of the conformation formed betweenthe ligand and the receptor. On the other hand, the expression of b3 receptor was analyzed by TLC in samples of RTEC treatedwith E 50 nM, 24 h in the presence and absence of Stx2 1 ng/ml). Docking analysis showed that EG presents an in silico 9-foldselectivity over GLS in comparison with Stx2, suggesting a strongeraf nity between E and LS. These results were according to TLCassay, which showed that E signi cantly inhibits b3 expressionat 24 h. Besides, RTEC cultures co-treated with E Stx2, showeda similar signi cant decrease in b3 expression. The incubation of RTEC with Stx2 alone maintained the same b3 expression levelas control non-treated cells. These results demonstrate that Stx2does not interfere with Eliglustat effect on Gb3 inhibition. Study sup-ported by PUE0041, CONICET.