in vitro evaluation of genetic variants and spatial transcriptomics of gonadal cells in patients with disorders of sex development

• Grant number: 25/08316-4
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
• Start date: July 01, 2025
• End date: June 30, 2027
• Field of knowledge: Biological Sciences - Genetics - Human and Medical Genetics
• Principal Investigator: Helena Fabbri Scallet
• Grantee: Felipe Rodrigues de Oliveira
• Host Institution: Centro de Biologia Molecular e Engenharia Genética (CBMEG). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil
• Associated research grant: 24/00196-7 - Investigating new processes in gonadal development, AP.JP

Abstract

Disorders of Sex Development (DSD) comprise a group of congenital conditions characterized by incomplete or atypical development of the genitalia and/or gonads, leading to discordance between genetic, gonadal, and phenotypic sex. As the same genetic variant has been associated with a wide phenotypic spectrum in unrelated individuals with DSD, it has been suggested that the interaction of two or more genes may contribute to the variability and, in some cases, to a more severe phenotype. This study proposes in vitro functional analyses of novel variants identified through next-generation sequencing (NGS), currently being conducted in parallel research projects. To assess the biological impact of these variants, different techniques will be employed, including western blotting and luciferase reporter assays.The second part of the project will involve the use of spatial transcriptomics to characterize gonadal cell populations. For this purpose, paraffin-embedded gonadal tissue samples will be sectioned and prepared on slides. These tissues will first undergo immunohistochemical analysis, followed by spatial transcriptomic profiling, with the goal of identifying morphological and molecular alterations in structures such as the Sertoli cell blood-testis barrier that may account for the different stages and presentations of DSD. This phase of the project will be conducted in collaboration with Professor Olaf Hiort's research group at the University of Lübeck, Germany, who is a team member of this proposal.