Molecular genetic diagnosis in patients with pituitary and gonadal developmental disorders and the use of models in vitro and in vivo to evaluate the functional effect of allelic variants identified by sequencing large scale

Abstract

The development of molecular biology techniques to evaluate initial candidate gene by Sanger sequencing technique and then by analysis of the translated from all exons of the human genome region (exoma) for sequencing on a large scale (SELE) led to increasing molecular diagnostic research in genetic diseases. The Sanger sequencing allowed to determine the molecular etiology in about 10% of patients with disorders of pituitary development and 20% of patients with disorders of sex development (DSD) followed at the Endocrinology clinic development Unit. The use of SELE allowed us to identify two new genes potentially involved in the development of pituitary (CDH2) and gonadal disorders (DHX37). According to the guidelines for investigating the causal variants in the genome sequence and its association with human disease, published by the National US Human Genome Research Institute, the strength of the correlation between the findings of an allelic variant and the phenotype of the patient is done with the genetic evidence and experimental level. In the first, it is fundamental the presence of variants in one gene in unrelated patients and in the second, the causal association of this variant in the functional context. Therefore, this project aims to analyze, By Sanger patients with Hypopituitarism in order to find CHD2 allelic variants and also by SELE, a greater number of patients with pituitary development disorders in the identification of new genes associated with hypopituitarism, and validate the functional impairment of these variants in vivo and in vitro models. (AU)