Detection of globotriaosylceramide (Gb3) in a HEK cell model

Teresa da Cunha e Silva Lopes Santos

Key information

Authors:

Teresa da Cunha e Silva Lopes Santos (Teresa Da Cunha e Silva Lopes Santos)

Supervisors:

Osvaldo Daniel Uchitel; Cláudia Alexandra Martins Lobato da Silva (Cláudia Lobato da Silva)

Published in

03/06/2023

Abstract

Fabry disease is a rare disorder within the spectrum of progressive lysosomal overload diseases. It is characterised by the deficient or absent lysosomal activity of α-galactosidase A, which results in the accumulation of globotriaosylceramide (Gb3). The accumulation of this glycosphingolipid causes some of the typical symptoms of Fabry's disease, as the appearance of angiokeratomas and neuropathic pain. In this project, the detection of Gb3 was studied using a model of HEK cells treated with drugs that produce the accumulation of Gb3 to then determine the optimal parameters that allow detection by microscopy. To determine the incubation time at which the intensity levels were higher, that is, there was a greater accumulation of Gb3, two different times were studied, a 30-minute incubation time and a 2-hour incubation time. The results obtained for Gb3, LysoGb3 and DGJ were also compared with each other. Increased Gb3 incorporation was observed in the treated cells, as well as increased accumulation in the plasma membrane, with higher values obtained for the two-hour incubation. Given its functional diversity, it would be interesting to continue studying the detection and location of Gb3, to contribute to the development of new and more effective therapies. Once it has been determined how and where this glycolipid accumulates, it would be beneficial to study how it is eliminated from the organ, its metabolization and degradation.