Sandra, MartinsThorsteinsdóttir, Sólveig, 1962-Santos, Bárbara Lopes dos2024-04-1720242023http://hdl.handle.net/10451/64383Tese de mestrado, Biologia Evolutiva e do Desenvolvimento , 2023, Universidade de Lisboa, Faculdade de CiênciasGaucher disease is a lysosomal disease caused by a mutation in the GBA1 gene. This gene encodes the lysosomal enzyme glucocerebrosidase (Gcase), which is responsible for the degradation of glucosylceramide into glucose and ceramide. Therefore, when a loss-of-function mutation occurs in the GBA1 gene, there is an abnormal accumulation of glucosylceramide in the lysosomes, responsible for the increase in volume of lysosomes. This disease presents a very wide spectrum of phenotypic manifestations, the most frequent of which is hepatosplenomegaly, and, in some situations, there may be involvement of the nervous system. Cilia are highly conserved and complex organelles that function as chemical and mechanical sensors of cells, presenting receptors for various signaling pathways such as Sonic Hedgehog (Shh) on their ciliary membrane. To try to understand which changes in genetic expression are associated with Gaucher disease, studies previously carried out in the laboratory of Prof.ª Dr.ª Maria Carmo-Fonseca compared, through RNA-seq, fibroblasts from apparently healthy individuals and individuals with Gaucher disease. Surprisingly, the bioinformatic analysis of these results showed changes in the alternative splicing pattern in a set of ciliary genes. The objective of this project was to try to understand how lysosomal dysfunctions culminate in changes in the regulation of genetic expression of primary cilium genes and whether these are reversible after treatment of Gaucher fibroblasts with recombinant Gcase, the enzyme used in the treatment of Gaucher patients. To this end, we morphologically analyzed the ciliary membrane and axoneme of primary cilia through the Arl13b and acetylated α-tubulin staining respectively, from Gaucher fibroblasts. After morphological analysis, we verified an increase in ciliary length in Gaucher fibroblasts compared to control one, which was reversed through treatment with Gcase. We also identified anomalies in the ciliary membrane and in the cilium axoneme of Gaucher fibroblasts, related to the pattern of acetylated α-tubulin that were slightly recovered upon treatment.engDoenças lisossomaisDoença de GaucherCílio primárioLisossomaSplicingTeses de mestrado - 2024master thesis203882130