Association of EEF1A1 and linc00993 expression with sperm morphology in teratozoospermia: an integrative in silico and in vitro study

Teratozoospermia (TZ) represents a significant factor in male infertility, and for many years, evaluation of sperm morphology has been the standard diagnostic method. This work investigates the molecular basis of TZ using a combined strategy that integrates computational analyses with experimental validation. Expression profiles of mRNA and lncRNA from individuals with TZ and from healthy subjects were examined using publicly available datasets from the NCBI Gene Expression Omnibus (GEO). We carried out differential expression analysis through GEO2R, then applied functional enrichment and protein–protein interaction network evaluations via the DAVID and STRING tools. Hub genes were identified and ranked based on their Degree centrality scores. To elucidate functional relationships among differentially expressed genes, RNA-binding proteins, and lncRNAs, we constructed interaction networks for lncRNA–mRNA and lncRNA–RBP interactions using Cytoscape. The expression patterns of EEF1A1 and linc00993, identified as principal candidates, were confirmed using quantitative real-time PCR. Notably, altered expression patterns of EEF1A1 and linc00993 were significantly associated with sperm morphology in teratozoospermia samples. These findings provide new insights into molecular alterations accompanying abnormal spermiogenesis and identify EEF1A1 and linc00993 as candidate molecular markers for future mechanistic and translational studies.