Integrated Analysis of Multiple Microarrays Based on Raw Data Identified Novel Gene Signatures in Recurrent Implantation Failure

Abstract

Background: Recurrent implantation failure (RIF) is an intricate complication following IVF-ET, which refers to the situation that good-quality embryos repeatedly fail to implant following two or more IVF cycles. Intrinsic molecular mechanisms underlying RIF have not yet been fully elucidated. With enormous improvement in high-throughput technologies, researchers screened biomarkers for RIF using microarray. However, the findings of published studies are inconsistent. An integrated study on the endometrial molecular determinants of implantation will help to improve pregnancy outcomes.

Objective: To identify robust differentially expressed genes (DEGs) and hub genes in endometrium associated with RIF, and to investigate the diagnostic role of hub genes in RIF.

Methods: Raw data from five GEO microarrays regarding RIF were analyzed. Integrated genetic expression analyses were performed using the Robust Rank Aggregation method to identify robust DEGs. Enrichment analysis and protein-protein interaction (PPI) analysis were further performed with the robust DEGs. Cytohubba was used to screen hub genes based on the PPI network. GSE111974 was used to validate the expression and diagnostic role of hub genes in RIF.

Results: 1532 Robust DEGs were identified by integrating four GEO datasets. Enrichment analysis showed that the robust DEGs were mainly enriched in processes associated with extracellular matrix remodeling, adhesion, coagulation, and immunity. A total of 18 hub genes (HMGCS1, SQLE, ESR1, LAMC1, HOXB4, PIP5K1B, GNG11, GPX3, PAX2, TF, ALDH6A1, IDH1, SALL1, EYA1, TAGLN, TPD52L1, ST6GALNAC1, NNMT) were identified. 10 of the 18 hub genes were significantly differentially expressed in RIF patients as validated by GSE111974. The 10 hub genes (SQLE, LAMC1, HOXB4, PIP5K1B, PAX2, ALDH6A1, SALL1, EYA1, TAGLN, ST6GALNAC1) were effective in predicting RIF with an accuracy rate of 85%, specificity rate of 100%, and sensitivity rate of 88.9%.

Conclusions: Our integrated analysis identified novel robust DEGs and hub genes in RIF. The hub genes were effective in predicting RIF and will contribute to the understanding of comprehensive molecular mechanisms in RIF pathogenesis.

Keywords: differentially expressed genes; hub gene; microarray; recurrent implantation failure; robust rank aggregation.