Causes and Genomic Approaches to Female Reproductive Failure. Doctoral Thesis

Abstract

Clinical recognition of the genetic causes of female reproductive failure using increasingly advancing genetic technologies to preserve patient safety and move towards personalized treatment application is a major challenge of reproductive medicine in the 21st century. The aim of this thesis was to demonstrate a reliable application of advanced genomic techniques in different stages of female reproductive failure in real-life clinical or research scenarios. Several genetic approaches were exploited – starting from the multifactor genetic testing of preimplantation embryos to select the ones free of inherited monogenic conditions and chromosomal aberrations, then following with the analysis of fetal material in case of early pregnancy loss using array comparative genomic hybridization and short tandem repeat analysis to exclude maternal cell contamination, and finally using next generation sequencing technology to analyze genetic landscape leading to preterm delivery in women with cervical insufficiency. The practical work described here was published as three scientific articles now forming three chapters of this thesis. Array comparative genomic hybridization combined with loci-specific genetic testing techniques allowed for a versatile and reliable analysis of preimplantation embryos to select the ones free of genetic conditions analyzed, and in combination with microsatellite analysis it also allowed to access the chromosomal causes of early pregnancy loss while reducing the misdiagnosis caused by maternal cell contamination. Next generation sequencing application allowed to identify the disruptive variants potentially contributive to the development of non-syndromic cervical insufficiency. Pathway enrichment analysis of variant genes from our cohort revealed an increased variation burden in genes playing roles in tissue mechanical and biomechanical properties. Literature analysis allowed to conclude that number of genes can be reliably attributed to female reproductive failure and an increasing number of genes form a pool of good candidates. In order to develop diagnostic gene panels and facilitate genetic advancement inclusion in the clinical practice of female reproduction, a standardized clinical gene-disease validity assessment of the identified genes has to be performed and best practice guidelines have to be composed.