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

Overview

Journal Biochem Genet

Date 2025 Feb 10

PMID 39928278

Authors

Fariba Ghodrati

Kazem Parivar

Iraj Amiri

Nasim Hayati Roodbari

Affiliations

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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.

References

Barcelo M, Mata A, Bassas L, Larriba S . Exosomal microRNAs in seminal plasma are markers of the origin of azoospermia and can predict the presence of sperm in testicular tissue. Hum Reprod. 2018; 33(6):1087-1098. PMC: 5972609. DOI: 10.1093/humrep/dey072. View

Blobel C . Functional processing of fertilin: evidence for a critical role of proteolysis in sperm maturation and activation. Rev Reprod. 2000; 5(2):75-83. DOI: 10.1530/ror.0.0050075. View

Bouhallier F, Allioli N, Lavial F, Chalmel F, Perrard M, Durand P . Role of miR-34c microRNA in the late steps of spermatogenesis. RNA. 2010; 16(4):720-31. PMC: 2844620. DOI: 10.1261/rna.1963810. View

Cabral-Pacheco G, Garza-Veloz I, Castruita-De la Rosa C, Ramirez-Acuna J, Perez-Romero B, Guerrero-Rodriguez J . The Roles of Matrix Metalloproteinases and Their Inhibitors in Human Diseases. Int J Mol Sci. 2021; 21(24). PMC: 7767220. DOI: 10.3390/ijms21249739. View

Chen H, Sampson N . Mediation of sperm-egg fusion: evidence that mouse egg alpha6beta1 integrin is the receptor for sperm fertilinbeta. Chem Biol. 1999; 6(1):1-10. DOI: 10.1016/S1074-5521(99)80015-5. View

Cho C . Testicular and epididymal ADAMs: expression and function during fertilization. Nat Rev Urol. 2012; 9(10):550-60. DOI: 10.1038/nrurol.2012.167. View

Comazzetto S, Di Giacomo M, Rasmussen K, Much C, Azzi C, Perlas E . Oligoasthenoteratozoospermia and infertility in mice deficient for miR-34b/c and miR-449 loci. PLoS Genet. 2014; 10(10):e1004597. PMC: 4199480. DOI: 10.1371/journal.pgen.1004597. View

Cooper T . Interactions between epididymal secretions and spermatozoa. J Reprod Fertil Suppl. 2000; 53:119-36. View

Daneshmandpour Y, Bahmanpour Z, Hamzeiy H, Mazaheri Moghaddam M, Mazaheri Moghaddam M, Khademi B . MicroRNAs association with azoospermia, oligospermia, asthenozoospermia, and teratozoospermia: a systematic review. J Assist Reprod Genet. 2020; 37(4):763-775. PMC: 7183035. DOI: 10.1007/s10815-019-01674-9. View

10.

Dickson D, Paulus J, Mensah V, Lem J, Saavedra-Rodriguez L, Gentry A . Reduced levels of miRNAs 449 and 34 in sperm of mice and men exposed to early life stress. Transl Psychiatry. 2018; 8(1):101. PMC: 5966454. DOI: 10.1038/s41398-018-0146-2. View

11.

Eto K, Sheppard D, Sehara-Fujisawa A, Zhang X, Takada Y . RGD-independent binding of integrin alpha9beta1 to the ADAM-12 and -15 disintegrin domains mediates cell-cell interaction. J Biol Chem. 2000; 275(45):34922-30. DOI: 10.1074/jbc.M001953200. View

12.

Frenette G, Legare C, Saez F, Sullivan R . Macrophage migration inhibitory factor in the human epididymis and semen. Mol Hum Reprod. 2005; 11(8):575-82. DOI: 10.1093/molehr/gah197. View

13.

Han C, Park I, Lee B, Jin S, Choi H, Kwon J . Identification of heat shock protein 5, calnexin and integral membrane protein 2B as Adam7-interacting membrane proteins in mouse sperm. J Cell Physiol. 2010; 226(5):1186-95. DOI: 10.1002/jcp.22444. View

14.

Hanwell M, Curtis D, Lonie D, Vandermeersch T, Zurek E, Hutchison G . Avogadro: an advanced semantic chemical editor, visualization, and analysis platform. J Cheminform. 2012; 4(1):17. PMC: 3542060. DOI: 10.1186/1758-2946-4-17. View

15.

Huang S, Zou X . A knowledge-based scoring function for protein-RNA interactions derived from a statistical mechanics-based iterative method. Nucleic Acids Res. 2014; 42(7):e55. PMC: 3985650. DOI: 10.1093/nar/gku077. View

16.

Jodati A, Pirouzpanah S, Maroufi N, Pezeshkian M, Safaie N, Bijanpour H . Different expression of Micro RNA-126, 133a and 145 in aorta and saphenous vein samples of patients undergoing coronary artery bypass graft surgery. J Cardiovasc Thorac Res. 2019; 11(1):43-47. PMC: 6477108. DOI: 10.15171/jcvtr.2019.07. View

17.

Jumper J, Evans R, Pritzel A, Green T, Figurnov M, Ronneberger O . Highly accurate protein structure prediction with AlphaFold. Nature. 2021; 596(7873):583-589. PMC: 8371605. DOI: 10.1038/s41586-021-03819-2. View

18.

Laskowski R . PDBsum new things. Nucleic Acids Res. 2008; 37(Database issue):D355-9. PMC: 2686501. DOI: 10.1093/nar/gkn860. View

19.

Li Y, Chen Y, Wu W, Li N, Hua J . MMPs, ADAMs and ADAMTSs are associated with mammalian sperm fate. Theriogenology. 2023; 200:147-154. DOI: 10.1016/j.theriogenology.2023.02.013. View

20.

Mariani V, Biasini M, Barbato A, Schwede T . lDDT: a local superposition-free score for comparing protein structures and models using distance difference tests. Bioinformatics. 2013; 29(21):2722-8. PMC: 3799472. DOI: 10.1093/bioinformatics/btt473. View