Masood Abu-Halima, Ulrike Fischer, Mohammad A Al Smadi, Nicole Ludwig, Anissa Acheli, Annika Engel, Hashim Abdul-Khaliq, Eckart Meese
{"title":"单精子 RNA 特征揭示了男性不育症的 MicroRNA 生物标记。","authors":"Masood Abu-Halima, Ulrike Fischer, Mohammad A Al Smadi, Nicole Ludwig, Anissa Acheli, Annika Engel, Hashim Abdul-Khaliq, Eckart Meese","doi":"10.1007/s10815-024-03264-w","DOIUrl":null,"url":null,"abstract":"<p><strong>Purpose: </strong>To investigate small RNA profiles in sperm, identify stable miRNA patterns unique to sperm, and assess the behavior of consistently expressed miRNAs in sperm from subfertile men compared to fertile controls.</p><p><strong>Methods: </strong>The small RNA profiles of single sperm from four proven fertile men were analyzed using Small RNA next-generation sequencing (NGS). Subsequently, a specific set of miRNAs was validated using RT-qPCR on additional sperm samples from 65 subfertile men from an infertility clinic and 30 proven fertile men.</p><p><strong>Results: </strong>Small RNA sequencing revealed a diverse range of sperm small RNA biotypes, including miRNAs. The mapped read percentage ranged from 22.19% for single sperm to 83.29% for enriched sperm samples used at different RNA concentrations. In single sperm, a smaller proportion of sequences were attributed to piRNAs (2.79%), miRNA (0.94%), tRNA (0.82%), and rRNA (0.47%) compared to enriched sperm samples, where piRNA (41.68%), tRNA (20.31%), miRNA (11.11%), and rRNA (6.54%) were observed. Distinct detection rates and a higher number of detected miRNAs were noted with enriched sperm samples compared to single sperm obtained using either a micromanipulator or microdissection systems. Among the identified miRNAs, 110 were consistently present in all samples. RT-qPCR revealed 15 miRNAs with increased expression and 5 miRNAs with decreased expression in sperm samples from subfertile men compared to proven fertile men. These differentially validated miRNAs were significantly correlated, either positively or negatively, with sperm count, motility, and morphology.</p><p><strong>Conclusion: </strong>The study extensively examines small RNAs in single sperm, identifying sperm-specific miRNAs that could serve as molecular markers to distinguish between subfertile and fertile men in clinical settings.</p>","PeriodicalId":15246,"journal":{"name":"Journal of Assisted Reproduction and Genetics","volume":" ","pages":"3119-3132"},"PeriodicalIF":3.2000,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11621271/pdf/","citationCount":"0","resultStr":"{\"title\":\"Single Sperm RNA signatures reveal MicroRNA biomarkers for male subfertility.\",\"authors\":\"Masood Abu-Halima, Ulrike Fischer, Mohammad A Al Smadi, Nicole Ludwig, Anissa Acheli, Annika Engel, Hashim Abdul-Khaliq, Eckart Meese\",\"doi\":\"10.1007/s10815-024-03264-w\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Purpose: </strong>To investigate small RNA profiles in sperm, identify stable miRNA patterns unique to sperm, and assess the behavior of consistently expressed miRNAs in sperm from subfertile men compared to fertile controls.</p><p><strong>Methods: </strong>The small RNA profiles of single sperm from four proven fertile men were analyzed using Small RNA next-generation sequencing (NGS). Subsequently, a specific set of miRNAs was validated using RT-qPCR on additional sperm samples from 65 subfertile men from an infertility clinic and 30 proven fertile men.</p><p><strong>Results: </strong>Small RNA sequencing revealed a diverse range of sperm small RNA biotypes, including miRNAs. The mapped read percentage ranged from 22.19% for single sperm to 83.29% for enriched sperm samples used at different RNA concentrations. In single sperm, a smaller proportion of sequences were attributed to piRNAs (2.79%), miRNA (0.94%), tRNA (0.82%), and rRNA (0.47%) compared to enriched sperm samples, where piRNA (41.68%), tRNA (20.31%), miRNA (11.11%), and rRNA (6.54%) were observed. Distinct detection rates and a higher number of detected miRNAs were noted with enriched sperm samples compared to single sperm obtained using either a micromanipulator or microdissection systems. Among the identified miRNAs, 110 were consistently present in all samples. RT-qPCR revealed 15 miRNAs with increased expression and 5 miRNAs with decreased expression in sperm samples from subfertile men compared to proven fertile men. These differentially validated miRNAs were significantly correlated, either positively or negatively, with sperm count, motility, and morphology.</p><p><strong>Conclusion: </strong>The study extensively examines small RNAs in single sperm, identifying sperm-specific miRNAs that could serve as molecular markers to distinguish between subfertile and fertile men in clinical settings.</p>\",\"PeriodicalId\":15246,\"journal\":{\"name\":\"Journal of Assisted Reproduction and Genetics\",\"volume\":\" \",\"pages\":\"3119-3132\"},\"PeriodicalIF\":3.2000,\"publicationDate\":\"2024-11-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11621271/pdf/\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of Assisted Reproduction and Genetics\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://doi.org/10.1007/s10815-024-03264-w\",\"RegionNum\":3,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"2024/9/23 0:00:00\",\"PubModel\":\"Epub\",\"JCR\":\"Q2\",\"JCRName\":\"GENETICS & HEREDITY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Assisted Reproduction and Genetics","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1007/s10815-024-03264-w","RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2024/9/23 0:00:00","PubModel":"Epub","JCR":"Q2","JCRName":"GENETICS & HEREDITY","Score":null,"Total":0}
Single Sperm RNA signatures reveal MicroRNA biomarkers for male subfertility.
Purpose: To investigate small RNA profiles in sperm, identify stable miRNA patterns unique to sperm, and assess the behavior of consistently expressed miRNAs in sperm from subfertile men compared to fertile controls.
Methods: The small RNA profiles of single sperm from four proven fertile men were analyzed using Small RNA next-generation sequencing (NGS). Subsequently, a specific set of miRNAs was validated using RT-qPCR on additional sperm samples from 65 subfertile men from an infertility clinic and 30 proven fertile men.
Results: Small RNA sequencing revealed a diverse range of sperm small RNA biotypes, including miRNAs. The mapped read percentage ranged from 22.19% for single sperm to 83.29% for enriched sperm samples used at different RNA concentrations. In single sperm, a smaller proportion of sequences were attributed to piRNAs (2.79%), miRNA (0.94%), tRNA (0.82%), and rRNA (0.47%) compared to enriched sperm samples, where piRNA (41.68%), tRNA (20.31%), miRNA (11.11%), and rRNA (6.54%) were observed. Distinct detection rates and a higher number of detected miRNAs were noted with enriched sperm samples compared to single sperm obtained using either a micromanipulator or microdissection systems. Among the identified miRNAs, 110 were consistently present in all samples. RT-qPCR revealed 15 miRNAs with increased expression and 5 miRNAs with decreased expression in sperm samples from subfertile men compared to proven fertile men. These differentially validated miRNAs were significantly correlated, either positively or negatively, with sperm count, motility, and morphology.
Conclusion: The study extensively examines small RNAs in single sperm, identifying sperm-specific miRNAs that could serve as molecular markers to distinguish between subfertile and fertile men in clinical settings.
期刊介绍:
The Journal of Assisted Reproduction and Genetics publishes cellular, molecular, genetic, and epigenetic discoveries advancing our understanding of the biology and underlying mechanisms from gametogenesis to offspring health. Special emphasis is placed on the practice and evolution of assisted reproduction technologies (ARTs) with reference to the diagnosis and management of diseases affecting fertility. Our goal is to educate our readership in the translation of basic and clinical discoveries made from human or relevant animal models to the safe and efficacious practice of human ARTs. The scientific rigor and ethical standards embraced by the JARG editorial team ensures a broad international base of expertise guiding the marriage of contemporary clinical research paradigms with basic science discovery. JARG publishes original papers, minireviews, case reports, and opinion pieces often combined into special topic issues that will educate clinicians and scientists with interests in the mechanisms of human development that bear on the treatment of infertility and emerging innovations in human ARTs. The guiding principles of male and female reproductive health impacting pre- and post-conceptional viability and developmental potential are emphasized within the purview of human reproductive health in current and future generations of our species.
The journal is published in cooperation with the American Society for Reproductive Medicine, an organization of more than 8,000 physicians, researchers, nurses, technicians and other professionals dedicated to advancing knowledge and expertise in reproductive biology.