Development of the quantification technique of exons for the detection of large deletions and insertions in the GBA gene for the diagnosis of Gaucher

• Grant number: 18/04401-3
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: June 01, 2018
• End date: December 31, 2018
• Field of knowledge: Biological Sciences - Genetics - Human and Medical Genetics
• Principal Investigator: João Bosco Pesquero
• Grantee: Jéssica Yumi Mitsuyassu
• Host Institution: Escola Paulista de Medicina (EPM). Universidade Federal de São Paulo (UNIFESP). Campus São Paulo. São Paulo , SP, Brazil

Abstract

Gaucher disease is caused by the dysfunction of the enzyme glycerebrosidase, causing a lysosomal accumulation, i.e., the lysosomes present in the macrophages and monocytes have accumulation of glucocerebroside. Such dysfunction induces the appearance of clinical symptoms which is classified as Gaucher Disease type 1, 2, 3.In Gaucher disease, there are mutations in the GBA gene alleles and a depletion of the enzyme glucocerebrosidase present in peripheral blood leukocytes. In this way, the diagnosis can be obtained by the morphological analysis obtained through the collection of bone marrow cells, or by the analysis of the enzymatic activity of glucocerebrosidase, in order to verify the depletion / absence of the enzyme and, lastly, by the molecular diagnosis, analyzing mutations present in the GBA gene.Gaucher treatment primarily comprises enzyme replacement, or substrate reduction. For better accuracy in the diagnosis of Gaucher, it is necessary to perform biochemical tests in conjunction with molecular tests. In order to do so, we can use the exons quantification technique (EQT), which aims to quantify the number of exons of the GBA gene, in order to analyze the presence of large deletions and insertions. Therefore, the objective of this work will be to standardize the EQT for Gaucher disease and then to perform a phenotype-genotype correlation, identifying pathogenic mutations and defining the diagnosis.