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Updated: Jun 5, 2025
In Vivo Functional Study of Disease-associated Rare Human Variants Using Drosophila
Published on: August 20, 2019
Whole-exome sequencing and Drosophila modelling reveal mutated genes and pathways contributing to human ovarian
Ismael Henarejos-Castillo1,2, Francisco José Sanz1,3, Cristina Solana-Manrique3,4
1IVI-RMA Global Research Alliance, IVI Foundation, Instituto de Investigación Sanitaria La Fe (IIS La Fe), Av. Fernando Abril Martorell 106, Valencia, 46026, Spain.
Background:
Ovarian failure (OF) is a multifactorial, complex disease presented by up to 1% of women under 40 years of age. Despite 90% of patients being diagnosed with idiopathic OF, the underlying molecular mechanisms remain unknown, making it difficult to personalize treatments for these patients in the clinical setting. Studying the presence and/or accumulation of SNVs at the gene/pathway levels will help describe novel genes and characterize disrupted biological pathways linked with ovarian failure.
Methods:
Ad-hoc case-control SNV screening conducted from 2020 to 2023 of 150 VCF files WES data included Spanish IVF patients with (n = 118) and without (n = 32) OF (< 40 years of age; mean BMI 22.78) along with GnomAD (n = 38,947) and IGSR (n = 1,271; 258 European female VCF) data for pseudo-control female populations. SNVs were prioritized according to their predicted deleteriousness, frequency in genomic databases, and proportional differences across populations. A burden test was performed to reveal genes with a higher presence of SNVs in the OF cohort in comparison to control and pseudo-control groups. Systematic in-silico analyses were performed to assess the potential disruptions caused by the mutated genes in relevant biological pathways. Finally, genes with orthologues in Drosophila melanogaster were considered to experimentally validate the potential impediments to ovarian function and reproductive potential.
Results:
Eighteen genes had a higher presence of SNVs in the OF population (FDR < 0.05). AK2, CDC27, CFTR, CTBP2, KMT2C, and MTCH2 were associated with OF for the first time and their silenced/knockout forms reduced fertility in Drosophila. We also predicted the disruption of 29 sub-pathways across four signalling pathways (FDR < 0.05). These sub-pathways included the metaphase to anaphase transition during oocyte meiosis, inflammatory processes related to necroptosis, DNA repair mismatch systems and the MAPK signalling cascade.
Conclusions:
This study sheds light on the underlying molecular mechanisms of OF, providing novel associations for six genes and OF-related infertility, setting a foundation for further biomarker development, and improving precision medicine in infertility.
Insights
This study identifies novel genes linked to ovarian failure (OF) and characterizes disrupted pathways, offering new insights into infertility causes. Findings pave the way for improved diagnostics and personalized treatments for women with OF.
Area of Science:
- Genetics and genomics
- Reproductive biology
- Molecular mechanisms of disease
Background:
- Ovarian failure (OF) affects up to 1% of women under 40.
- Idiopathic OF has unknown molecular causes, hindering personalized treatment.
- Identifying genetic and pathway disruptions is crucial for understanding OF.
Purpose of the Study:
- To identify novel genes and disrupted biological pathways associated with ovarian failure.
- To characterize the molecular underpinnings of idiopathic OF.
- To lay the groundwork for improved diagnostics and precision medicine in infertility.
Main Methods:
- Case-control study analyzing whole exome sequencing (WES) data from 118 OF patients and 32 controls.
- Prioritization of single nucleotide variants (SNVs) based on deleteriousness, frequency, and population differences.
- In-silico analysis of gene-pathway disruptions and experimental validation in Drosophila melanogaster.
Main Results:
- Eighteen genes showed a higher SNV frequency in the OF cohort (FDR < 0.05).
- Six genes (AK2, CDC27, CFTR, CTBP2, KMT2C, MTCH2) were newly associated with OF and impaired fertility in Drosophila.
- Disruption of 29 sub-pathways within four signaling pathways, including meiosis, necroptosis, DNA repair, and MAPK signaling, was predicted.
Conclusions:
- This research elucidates molecular mechanisms underlying OF, identifying novel gene associations.
- The findings provide a foundation for developing biomarkers for OF-related infertility.
- This study advances precision medicine approaches for infertility treatment.
