Elevated LncRNA LINC00966 Suppresses PDE2A to Impair Sperm Motility in Asthenozoospermia

Abstract

Emerging evidence indicated that long non-coding RNAs (lncRNAs) play critical roles in mammalian reproductive physiology, yet their roles in asthenozoospermia (AS), a major contributor to male infertility, remain elusive. Here, we identify LINC00966 as a candidate lncRNA associated with impaired sperm function. LINC00966 expression was significantly elevated in semen from AS patients and negatively correlated with sperm concentration, total motility, and progressive motility. In mouse testes, Linc00966 was detected in spermatogonia, spermatocytes and spermatids with predominant cytoplasmic localization as well as in GC-2spd cells. Furthermore, Linc00966 overexpression reduced cell viability and migration and increased intracellular reactive oxygen species (ROS), whereas knockdown enhanced viability and migration and lowered ROS in GC-2spd cells. Bulk RNA-seq analysis revealed that the phosphodiesterase 2A (Pde2a), a key cGMP/cAMP-hydrolyzing enzyme, were upregulated when Linc00966 was knockdown. The GO enrichment and KEGG analysis of differentially expressed genes suggested the cyclic guanosine monophosphate (cGMP) process were enriched. Further validation found that Linc00966 suppressed Pde2a. In clinical semen samples, LINC00966 and PDE2A expression levels were inversely correlated, and both were closely associated with semen quality parameters. Collectively, these findings implicated LINC00966 and its downstream target PDE2A as potential pathogenic regulators of sperm motility in AS. By linking lncRNA-mediated modulation of cGMP signaling and oxidative stress to spermatogenic cell function, our results demonstrated a plausible signaling axis underlying AS and suggests LINC00966/PDE2A as promising candidates for biomarker development and therapeutic investigation in male infertility.