Background: Intrauterine insemination (IUI) is often the first-line treatment for unexplained infertility. β-Microsemino protein (MSMB) is an abundant protein in seminal plasma that has an inhibitory effect on spontaneous acrosome reaction.

Objective: The present study aimed to evaluate MSMB gene variations and protein expression on IUI success rate in unexplained infertile men.

Materials and Methods: A case-control study was performed on 100 unexplained infertile Iranian men referred to the Royan Institute, Tehran, Iran for IUI (50 men with IUI positive result [IUI+], and 50 men with IUI negative result [IUI-]). Couples with female infertility factors (such as hormonal disorders, infrequent menstrual period, abnormality in uterus, fallopian tubes, or ovaries) and men with infections of the male accessory glands, hypogonadotropic hypogonadism, clinical varicocele, retractile testis, genital trauma, drug use, or concurrent hormonal treatment Y chromosome microdeletions, and abnormal karyotype were excluded from the study. The polymerase chain reaction sequencing was performed for the promoter and the coding regions of MSMB functional domains. To study the protein expression, the total protein of sperm was extracted, and sandwich enzyme-linked immunosorbent assay was performed.

Results: 4 variations were detected (rs12770171, rs10993994, rs2075894, and rs4517463). None of them showed significant differences between the IUI+ and IUI-groups. The mean value of protein expression did not show any differences between the groups.

Conclusion: In conclusion, there is no association between genetic variations of promoter and coding regions of MSMB functional domains as well as its expression with IUI success in unexplained infertile men.

 Key words: MSMB, Beta microseminoprotein, Infertility, Male infertility.

[1] Dugule DS-G, Gana ED, Paul J. Causes, effects and management of infertility among reproductive women in Karim Lamido Local Government Area of Taraba State-Nigeria. Texas J Med Sci 2023; 16: 36–49.

[2] Panner Selvam MK, Agarwal A, Pushparaj PN, Baskaran S, Bendou H. Sperm proteome analysis and identification of fertility-associated biomarkers in unexplained male infertility. Genes 2019; 10: 522.

[3] Kamath MS, Rikken JF, Bosteels J. Does laparoscopy and hysteroscopy have a place in the diagnosis of unexplained infertility? Semin Reprod Med 2020; 38: 29–35.

[4] Kawale SP, Ambad R, Jha RK, Jha RK, Gaikwad A. Role of artificial intelligence in ICSI (intra-cytoplasmic sperm injection) & IUI (intra-uterine insemination). E3S Web of Conferences; 2024 January 11–12: India.

[5] Erdem M, Erdem A, Firat Mutlu M, Ozisik S, Yildiz S, Guler I, et al. The impact of sperm morphology on the outcome of intrauterine insemination cycles with gonadotropins in unexplained and male subfertility. Eur J Obstet Gynecol Reprod Biol 2016; 197: 120–124.

[6] Aboutorabi R, Zamani S, Zarrin Y, Mostafavi FS. A survey on main semen parameters in natural pregnancy and intrauterine insemination: Are there any significant differences? Am J Mens Health 2018; 12: 617–623.

[7] Atasever M, Kalem MN, Hat??rnaz S, Hat??rnaz E, Kalem Z, Kalayl??oglu Z. Factors affecting clinical pregnancy rates after IUI for the treatment of unexplained infertility and mild male subfertility. J Turk Ger Gynecol Assoc 2016; 17: 134–138.

[8] Siu KK, Serrão VHB, Ziyyat A, Lee JE. The cell biology of fertilization: Gamete attachment and fusion. J Cell Biol 2021; 220: e202102146.

[9] Tumova L, Zigo M, Sutovsky P, Sedmikova M, Postlerova P. The ubiquitin-proteasome system does not regulate the degradation of porcine ??- microseminoprotein during sperm capacitation. Int J Mol Sci 2020; 21: 4151.

[10] Anklesaria JH, Mhatre DR, Mahale SD. Structural and molecular biology of PSP94: Its significance in prostate pathophysiology. Front Biosci 2018; 23: 535–562.

[11] Grande G, Vincenzoni F, Mancini F, Barrachina F, Giampietro A, Castagnola M, et al. Quantitative analysis of the seminal plasma proteome in secondary hypogonadism. J Clin Med 2019; 8: 2128.

[12] Rostami Chayjan M, Sabbaghian M, Alikhani M, Sahraneshin Samani F, Salman Yazdi R, Almadani SN, et al. [Relation of ??-defensin 126 gene (DEFB126) alteration and protein expression with unexplained male infertility in Iranian population]. J Arak Univ Med Sci 2014; 17: 31–40. (in Persian)

[13] Bu Y, Wang P, Li S, Li L, Zhang S, Wei H. Semen protein CRISP3 promotes reproductive performance of boars through immunomodulation. Int J Mol Sci 2024; 25: 2264.

[14] El Atab O, Kocabey AE, Asojo OA, Schneiter R. Prostate secretory protein 94 inhibits sterol binding and export by the mammalian CAP protein CRISP2 in a calciumsensitive manner. J Biol Chem 2022; 298: 101600.

[15] World Health Organization. WHO laboratory manual for the examination and processing of human semen. Switzerland: World Health Organization Press; 2021.

[16] Dairawan M, Shetty PJ. The evolution of DNA extraction methods. Am J Biomed Sci Res 2020; 8: 39–45.

[17] Mendivelso González DF, Sánchez Villalobos SA, Ramos AE, Montero Ovalle WJ, Serrano López ML. Single nucleotide polymorphisms associated with prostate cancer progression: A systematic review. Cancer Invest 2024; 42: 75–96.

[18] Dariane C, Clairefond S, Péant B, Communal L, Thian Z, Ouellet V, et al. High keratin-7 expression in benign peri-tumoral prostatic glands is predictive of bone metastasis onset and prostate cancer-specific mortality. Cancers 2022; 14: 1623.

[19] Nasimi H, Madsen JS, Zedan AH, Malmendal A, Osther PJS, Alatraktchi FAa. Protein biomarker detection in prostate cancer: A comprehensive review of electrochemical biosensors. Sensors and Actuators Reports 2023; 6: 100168.

[20] Carvalho AdL, Pinto SA, Santos WGd. CRISP3 glycoprotein: A good biomarker for prostate cancer? J Brasil Patol Med Lab 2021; 57: e3222021.

[21] Fu S, Huang Y-L, Luan T, Li N, Wang H-F, Wang JS. A meta-analysis of influence of MSMB promoter rs10993994 polymorphisms on prostate cancer risk. Eur Rev Med Pharmacol Sci 2019; 23: 9295–9303.

[22] Tian P, Zhong M, Wei G-H. Mechanistic insights into genetic susceptibility to prostate cancer. Cancer Lett 2021; 522: 155–163.

[23] Frapsauce C, Pionneau C, Bouley J, Delarouziere V, Berthaut I, Ravel C, et al. Proteomic identification of target proteins in normal but nonfertilizing sperm. Fertil Steril 2014; 102: 372–380.

[24] Agarwal A, Selvam MKP, Baskaran S. Proteomic analyses of human sperm cells: Understanding the role of proteins and molecular pathways affecting male reproductive health. Int J Mol Sci 2020; 21: 1621.

[25] Emami NC, Kachuri L, Meyers TJ, Das R, Hoffman JD, Hoffmann TJ, et al. Association of imputed prostate cancer transcriptome with disease risk reveals novel mechanisms. Nat Commun 2019; 10: 3107.

[26] Trujillo-Cáceres SJ, Torres-Sánchez L, Burguete-García AI, Orihuela YCO, Vázquez-Salas RA, Álvarez-Topete E, et al. Contribution of MSMB promoter region gene polymorphism to early-onset prostate cancer risk in Mexican males. Oncotarget 2019; 10: 738.

[27] Labani M, Beheshti A, Argha A, Alinejad-Rokny H. A Comprehensive investigation of genomic variants in prostate cancer reveals 30 putative regulatory variants. Int J Mol Sci 2023; 24: 2472.

[28] Samtal Ch, El Jaddaoui I, Hamdi S, Bouguenouch L, Ouldim K, Nejjari C, et al. Review of prostate cancer genomic studies in Africa. Front Genet 2022; 13: 911101.

[29] Desai TA, Hedman AK, Dimitriou M, Koprulu M, Figiel S, Yin W, et al. Identifying proteomic risk factors for overall, aggressive and early onset prostate cancer using Mendelian randomization and tumor spatial transcriptomics. medRxiv 2023. (preprint)

[30] Jodar M, Soler-Ventura A, Oliva R, Molecular Biology of Reproduction and Development Research Group. Semen proteomics and male infertility. J Proteomics 2017; 162: 125–134.