Estudo da homeostase lisossômica e caracterização de fibroblastos de pacientes com doença de Fabry em cultura celular

• Autor(a) principal: Gomez, Nadia Aylin [UNIFESP]
• Data de Publicação: 2017
• Tipo de documento: Dissertação
• Idioma: por
• Título da fonte: Repositório Institucional da UNIFESP
• Texto Completo: https://sucupira.capes.gov.br/sucupira/public/consultas/coleta/trabalhoConclusao/viewTrabalhoConclusao.jsf?popup=true&id_trabalho=5074612; http://repositorio.unifesp.br/handle/11600/50428
• topic: Fabry Disease; Lysosomal Storage Disorder; Lysosomal Homeostasis; Lysosomal Calcium; Mtorc1; Doença de Fabry; Genética; Doenças por armazenamento dos lisossomos; Serina-treonina quinases TOR

Fabry disease is one of the lysosomal storage diseases which has a X-linked recessive inheritance. A mutation in GLA gene, causes α-galactosidase A (α-Gal A) deficiency, that with the crescent accumulation of globotriaosylceramide (Gb3) results in signs and symptoms. This is a chronic disease, and the vascular endothelial cells are the most compromised, leading to a multiorgan disease, with some neurologic manifestations that can cause premature death. Lysosomes are the principal cytosolic organelles affected in Fabry disease, and their normal function is necessary to several vital processes. They act in autophagy, endocytosis, exocytosis and phagocytosis among other functions. The regulation of lysosomal homeostasis is very important and recently it was demonstrated that the lysosomal-nucleus signaling participated of this process. The complete impact of the enzymatic deficiency in lysosomes with Fabry disease is still unknown. For this reason, the aim of our study is to evaluate Ca2+, mTOR and TFEB, the principal players of the machinery that regulates the autophagy and lysosomal biogenesis, in fibroblasts of Fabry disease patients. In this work, we characterized fibroblasts of Fabry disease patients, observing the deficiency of α-Gal A and the accumulation of Gb3. Also, it was verified an impaired lysosomal function in the enzymatic activity of β-Glucosidase, acid α-Glucosidase and α-L-Iduronidase. The biochemical parameters, that participated of the lysosomal homeostasis regulation and the cytoplasmic pH, were evaluated, but did not show significant differences, which suggests that the cells maintain the lysosomal regulation in equilibrium and cellular homeostasis, too. It was verified that the Ca2+cytoplasmatic (Ca2+cyt) e o Ca2+endoplasmic reticulum (Ca2+ret), in male and female patients cell cultures were modified, which, in turn, indicates alterations in mechanisms dependent on this ion. In conclusion, fibroblast of Fabry disease patients shows an alteration in the lysosomal enzymes evaluated, as well as, in the concentration of Ca2+cyt and Ca2+ret which seems do not to affect the lysosomal homeostasis regulation, or even the cellular equilibrium.