In vitro models for the study of the neuropathophysiology of Fabry disease

• Grant number: 23/14694-6
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• Start date: April 01, 2025
• End date: March 31, 2027
• Field of knowledge: Biological Sciences - Genetics - Human and Medical Genetics
• Principal Investigator: João Bosco Pesquero
• Grantee: Priscila Nicolicht de Amorim
• Host Institution: Escola Paulista de Medicina (EPM). Universidade Federal de São Paulo (UNIFESP). Campus São Paulo. São Paulo , SP, Brazil
• Associated research grant: 20/14635-1 - Modeling of monogenic diseases for physiopathological studies and pharmacological tests using specialized cells derived from iPSCs, AP.TEM

Abstract

Fabry disease (FD) is an inborn error of metabolism caused by mutations in the GLA gene located on the X chromosome. Defects in this gene lead to the deficiency of ±-galactosidase A (±-Gal A), an enzyme responsible for degrading glycosphingolipids, mainly globotriaosylceramide (Gb3). ±-Gal A is present in all tissues, and its deficiency results in a progressive multisystem disease. FD covers a broad clinical phenotypic spectrum, mainly affecting cardiac, renal, gastrointestinal and cerebrovascular systems. Cerebrovascular and neuropsychiatric symptoms are described in patients with FD, however, the pathophysiological basis of FD in the central nervous system is poorly understood. We propose to develop an in vitro model to study the neurophysiopathology of FD through the use of induced pluripotent stem cells (iPSCs). Firstly, we will use the CRISPR/Cas9 technique to edit the GLA gene of iPSCs; These cells will be characterized for the efficiency of mutation induction, absence of off-targets and pluripotency potential, as well as for the deficiency of ±-Gal A enzymatic activity. The cells will then be differentiated into astrocytes and brain endothelial cells, being that Gb3 accumulation as well as cell survival will be evaluated. Finally, transcriptomics and proteomics will be performed on both edited and wild-type cells, in order to analyze the possible impact of ±-Gal A deficiency on the expression of mRNAs and proteins that may be related to cerebrovascular and neuropsychiatric changes found in patients. with DF. With this project, we will take an important step in the studies of the neurophysiopathology of FD, a topic that has been little explored but is of vital importance for the treatment of patients.