Exploring miR-34a, miR-449, and ADAM2/ADAM7 Expressions as Potential Biomarkers in Male Infertility: A Combined In Silico and Experimental Approach.

Abstract

miR-34a and miR-449 are key miRNAs involved in sperm function and male fertility, with their dysregulation potentially contributing to male infertility. ADAM proteins, specifically ADAM2 and ADAM7, are also implicated in sperm function. This study investigates the interactions between miR-34a, miR-449, and ADAM2/ADAM7, exploring their roles in male infertility through both experimental analyses and molecular docking. In this case-control study, 15 infertile males and 15 healthy controls were included. Gene expression levels of miR-34a, miR-449, and SOX30 were measured using real-time PCR, while protein levels of ADAM7 and ADAM2 in sperm were assessed through western blotting. Additionally, molecular docking was performed to analyze the binding affinities between miR-34a/miR-449 and ADAM2/ADAM7, with docking scores and confidence levels evaluated. Expression levels of ADAM7 and ADAM2 proteins in sperm from the infertile group showed significant differences compared with the control group (P ≤ 0.05). A significant difference was observed in the expression of miR-449, miR-34a, and SOX30 genes between the control and infertile groups (P < 0.05). A significant correlation between miR-34a expression, ADAM7 protein expression, and sperm morphology was observed. However, no statistically significant correlation was found between miR-34a expression and sperm motility, sperm count, blastocyst, or embryo rates in ICSI and IVF (P ≥ 0.05). Molecular docking and dynamics studies revealed strong interactions between miR-34a/miR-449 and ADAM proteins. The ADAM7/miR-34a complex showed the highest binding affinity with a docking score of - 372.40 and a confidence score of 0.9884, followed by ADAM7/miR-449. Hydrogen bond analysis indicated stable binding, with 9 bonds for ADAM2/miR-34a and 7 for ADAM7/miR-34a. These interactions suggest a significant role in regulating sperm morphology and function.miR-34a, miR-449, ADAM7, and ADAM2 protein expression appear to be involved in the molecular mechanisms of male infertility. These parameters show potential as biomarkers in assisted reproductive technology techniques, particularly by influencing sperm morphology and function.