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MicroRNA and Male Infertility: A Potential for Diagnosis
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Abstract
MicroRNAs (miRNAs) are small non-coding single stranded RNA molecules that are physiologically produced in eukaryotic cells to regulate or mostly down-regulate genes by pairing with their complementary base-sequence in related mRNA mol-ecules in the cytoplasm. It has been reported that other than its function in many physiological cell processes, dysregulation of miRNAs plays a role in the develop-ment of many diseases. In this short review, the association between miRNAs and some male reproductive disorders is surveyed. Male factor Infertility is a devas-tating problem from which a notable percentage of couples suffer. However, the molecular mechanism of many infertility disorders has not been clearly elucidated. Since miRNAs have an important role in numerous biological cell processes and cellular dysfunctions, it is of interest to review the related literature on the role of miRNAs in the male reproductive organs. Aberrant expression of specific miRNAs is associated with certain male reproductive dysfunctions. For this reason, assess-ment of expression of such miRNAs may serve as a suitable molecular biomarker for diagnosis of those male infertility disorders. The presence of a single nucleotide polymorphism (SNP) at the miRNAs’ binding site in its targeted mRNA has been reported to have an association with idiopathic male infertility. Also, a relation with male infertility has been shown with SNP in the genes of the factors necessary for miRNA biogenesis. Therefore, focusing on the role of miRNAs in male reproduc-tive disorders can further elucidate the molecular mechanisms of male infertility and generate the potential for locating efficient biomarkers and therapeutic agents for these disorders.
Keywords: miRNA, Spermatogenesis, Male Infertility, Biomarker, Single Nucleotide Poly-morphism
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Introduction
MicroRNAs (miRNAs) are small non-coding single stranded RNA molecules of approxi-mately 22 nucleotides that are physiological-ly produced in eukaryotic cells (1). After be-ing expressed in the nucleus, miRNA plays a role in regulation of gene expression by pair-ing with its complementary base-sequence of the targeted mRNA molecule in the cytoplasm. This usually leads to gene silencing through
Fig 1: Biogenesis of miRNA: RNA polymerase II transcribes miRNA gene to produce Pri-miRNA having a stem loop structure. Drosha (a class 2 ribonuclease III) and its RNA-binding partner, DGCR8*, cut the stem loop structure to make pre-miRNA. Exportin 5 transports the pre-miRNA from the nucleus to cytoplasm. In the cytoplasm, Dicer (a RNase III endonuclease) and its RNA-binding partner, TRBP**, cleave pre-miRNA to generate a short double-stranded RNA molecule named miRNA duplex. One strand of this molecule is incorporated into the miRNA-induced silencing complex (miRISC) containig protein Argonaute-2 (AGO2) as a catalytic component to regulate the targeted mRNA. The mechanism of targeted mRNA silencing could be either translational repression or mRNA cleavage.
miRNA; MicroRNA, * DGCR8; DiGeorge syndrome critical region gene and ** TRBP; Transactivating region binding protein.
Aside from the physiological regulation of gene expression in multiple biological process-es such as cell cycle control and differentiation (9), cell growth and apoptosis (10) and embryo development (11), the role of miRNAs in de-velopment of many human diseases has been studied and reported. Mutation, dysfunction of biogenesis and dysregulation of miRNAs and their targets may lead to various diseases such as cardiovascular diseases (12), cancers (13), schizophrenia (14), renal function disorders (15), psoriasis (16), muscular disorders (17) and diabetes (18). A comprehensive database of whole miRNA-disease association, miRNA
GLVHDVHVSHFWUXPZLGWK'6:PL51$FRQVHU -vation and miRNA function has been published
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focus on male infertility with the intent to con-duct a literature review of the role of miRNAs
in this field.
Infertility is a problem in 10-15% of couples worldwide (19). Approximately, 50% of the in-fertility cases are attributed to male factors and 60-5% of these cases are idiopathic (20). Of note, the molecular mechanisms of many male infertility disorders are not clear (20). The aim of this review is to pursue the relation between miRNAs and male infertility.
miRNA and spermatogenesis
The presence of miRNA in testis has been verified a few years ago (21). Later, researchers focused on finding the function of miRNAs in germinal tract physiology. Table 1 shows a brief description of the chronological order of dis-coveries related to miRNAs in testis molecular physiology [for additional information please
Table 1: Discoveries related to microRNAs (miRNAs) in the male reproductive system
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microarray, quantitative real-time PCR
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Potential of extracellular miRNAs as biomarkers for infertility
In addition to the presence of miRNA at the cel-lular level, miRNAs have also been reported to
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(34), saliva (35, 36), vaginal secretions, menstrual blood and semen (36). Aberrant expression of ex-tracellular miRNA is attributed to different disor-ders (37). Altered expression of miRNAs, as with
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(38-41) has been recently reported in male infertil-ity disorders, hence the possibilinfertil-ity of locating po-tential miRNA biomarkers in male infertility has been proposed.
7KH ¿UVW UHSRUW RI DQ DOWHUDWLRQRI PL51$ H[ -pression in the testis of patients with non-obstruc-tive azoospermia (NOA) has been reported by Lian et al. (42). Of note, that NOA which has a tes-tis-origin, presents a diverse range of defects from hypospermatogenesis and sperm maturation arrest to Sertoli-cell-only-syndrome. Hence, assessment of differential expressions of miRNAs in these in-fertile individuals is a prerequisite.
Wang et al. (43) examined pooled semen sam-ples obtained from infertile men and compared the results with normal fertile individuals as controls. They found alterations in miRNA pro-files by Solexa Sequencing (a sequencing meth-od based on reversible dye-terminators tech-nology and engineered polymerases) in both azoospermia and asthenozoospermia. These au-thors considered a cut of value of 50-fold higher or lower expression as significant, which was further validated by real time RT-qPCR for 7 miRNAs (miR-34c-5p, miR-122, miR-146b-5p, 181a, 374b, 509 -5p, and miR-513a-5p). The level of these 7 miRNAs was significantly lower in azoospermia and higher in asthenozoospermia compared to the control. Another noticeable finding of these authors was the stability of the above miRNAs to different conditions. They attributed this phenomenon to the small size of the miRNAs and their ability to bind with complex organic molecules in cell-free semen samples. Finally, they proposed that these 7 miRNAs might have confirmative mo-lecular diagnostic value for male infertility.
In this regard, Wu et al. (44) evaluated the pat-tern of expression related to miR-I9b and let-7a in
idiopathic infertile individuals with NOA or oligo-zoospermia by quantitative RT-PCR. They showed that these two miRNAs distinctively expressed at higher levels in infertile cases compared with fer-tile individuals. Consequently, they concluded that miR-I9b and let-7a were good diagnostic molecu-lar biomarkers for idiopathic infertile cases with NOA or oligozoospermia.
Single nucleotide polymorphism (SNP) and func-tion of miRNA in spermatogenesis
Considering the diverse role of miRNA in sper-matogenesis, therefore any polymorphism in re-lated genes might lead to infertility. Zhang et al.
ZHUHWKH¿UVWUHVHDUFKHUVZKRUHSRUWHGDQDV -sociation between miRNA-binding site single nu-cleotide polymorphism (SNP) and idiopathic male infertility. They systematically surveyed all SNPs in the 3´ UTR of 140 mammal spermatogenesis-related genes and observed that some SNPs within miRNA-binding sites might be related to idiopath-ic male infertility.
Among 140 surveyed genes, 6 SNPs (two in CYP19, Serpina5 and four in CGA, CPEB1, and CPEB2), involved in down regulation of gene expression in spermatogenesis and meio-sis and were predicted to have possibility for altering the binding affinity of miRNA by using bioinformatically specialized algorithms (Pic-tar, miRanda, Targetscan, and RNAhybrid). The effectiveness of these SNPs in male infertility was analyzed in a case-control study using PCR followed by restriction digestion of the ampli-fied fragments for genotyping. They realized that T substituted for A in rs6631 which lead to diminished binding ability of miR-1302 to its binding site in CGA in vitro, with a subsequent
overexpression of CGA. Therefore an associa-tion between this special SNP and male infertil-ity was concluded.
Recently Qin et al. (46) evaluated the
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Conclusion
Considering limited number of studies per-formed in this field, little has been clarified about miRNA-mediated gene regulation in spermatogenesis. In addition, there are few numbers of published studies that focus on the role of miRNAs in male infertility. This review has aimed to elaborate the role of miRNA in the male reproductive system. Any disorder or fail-ure in this system can result in male infertility. Therefore, a dysfunction in miRNA processing as
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and infertility. Other anomalies such as dysregula-tion in expression of certain miRNAs or SNP in the miRNA binding site or SNP in the genes involved in biogenesis of miRNA can be related to infer-tility. Hence, in this regard, scientists suggest that assessment of miRNA may have future diagnostic value and shed more light on the molecular mecha-nisms of male infertility. Thus, new paths can be opened for future treatments of male infertility or even in the design of new contraceptive drugs.
Acknowledgements
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