Fabry disease (FD) is an inborn error of the metabolism caused by mutations in the GLA gene (GLA; OMIM * 300644), located on the X chromosome. Defects in GLA lead to ±-galactosidase A deficiency (alpha-Gal A; .1.22), responsible for degrading glycosphingolipids, mainly globotriaosylceramide (Gb3). The ± -Gal A is present in all tissues; the deficiency of this enzyme results in a progressive multisystemic disease. FD encompasses a broad clinical phenotype spectrum, mainly affecting cardiac, renal, gastrointestinal and cerebrovascular systems. Neuropsychiatric symptoms are repeatedly described in patients with FD; however, the pathophysiological bases of FD in the central nervous system are poorly understood and it is unclear whether psychiatric symptoms are caused by the disease or secondary. We propose here to develop an in vitro model to study the pathophysiology of FD in neurons. First, we will use the CRISPR Cas9 technique to edit SH-SY5Y cells, a lineage capable of differentiating into neurons. These cells will be characterized by the efficiency of the mutations induction in the GLA gene and absence of off-targets, as well as for the deficiency of the enzymatic activity of the ±-Gal A. Then, the cells will be differentiated in neurons; the Gb3 accumulation as well as the cellular survival will be evaluated. Finally, we will perform the transcriptome of the edited and wild type cells, in order to analyze the possible impact of ±-Gal A deficiency on neurotransmitters, receptors, as well as on the expression of mRNAs related to proteins that are altered in psychiatric disorders.
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