Eliglustat inhibits glucosylceramide synthase and globotriaosylceramide expression without interacting with Shiga toxin 2

SANCHEZ DAIANA; CASAL JUAN; IBARRA, CRISTINA; AMARAL MARÍA M; SILBERSTEIN CLAUDIA

Modalidad Virtual

Congreso; Reunión de Sociedades de Biociencias 2021; 2021

SAIC. SAI. AAFE. NANOMED

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 (GLS), the first step of glycosphingolipidspathway, also inhibits Gb3 expression and preventsthe cytotoxic effects of Shiga toxin type 2 (Stx2) in human corticalrenal tubular epithelial cells (HRTEC) primary cultures. The aim ofthis work was to clarify the mode of interaction of EG in the activesite of GLS and its possible interaction with Stx2 and Gb3, andcompare to Gb3 expression in HRTEC treated with EG and Stx2.For this, a computational procedure called molecular docking wascarried out with Smina software and Gibbs free energy was calculatedfor determining the stability of the conformation formed betweenthe ligand and the receptor. On the other hand, the expression ofGb3 receptor was analyzed by TLC in samples of HRTEC treatedwith EG (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 strongeraffinity between EG and GLS. These results were according to TLCassay, which showed that EG significantly inhibits Gb3 expressionat 24 h. Besides, HRTEC cultures co-treated with EG+Stx2, showeda similar significant decrease in Gb3 expression. The incubation ofHRTEC with Stx2 alone maintained the same Gb3 expression levelas control non-treated cells. These results demonstrate that Stx2does not interfere with Eliglustat effect on Gb3 inhibition. Study supportedby PUE0041, CONICET.