ESX1 gene as a potential candidate responsible for male infertility in nonobstructive azoospermia
Abstract Infertility is a problem that affects approximately 15% of couples, and male infertility is responsible for 40–50% of these cases. The cause of male infertility is still poorly diagnosed and treated. One of the prominent causes of male infertility is disturbed spermatogenesis, which can lea...
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2023-10-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-023-43854-9 |
| _version_ | 1797453331653722112 |
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| author | Agnieszka Malcher Zuzanna Graczyk Hermann Bauer Tomasz Stokowy Andrea Berman Mikołaj Smolibowski Dominika Blaszczyk Piotr Jedrzejczak Alexander N. Yatsenko Maciej Kurpisz |
| author_facet | Agnieszka Malcher Zuzanna Graczyk Hermann Bauer Tomasz Stokowy Andrea Berman Mikołaj Smolibowski Dominika Blaszczyk Piotr Jedrzejczak Alexander N. Yatsenko Maciej Kurpisz |
| author_sort | Agnieszka Malcher |
| collection | DOAJ |
| description | Abstract Infertility is a problem that affects approximately 15% of couples, and male infertility is responsible for 40–50% of these cases. The cause of male infertility is still poorly diagnosed and treated. One of the prominent causes of male infertility is disturbed spermatogenesis, which can lead to nonobstructive azoospermia (NOA). Whole-genome sequencing (WGS) allows us to identify novel rare variants in potentially NOA-associated genes, among others, in the ESX1 gene. The aim of this study was to activate the ESX1 gene using CRISPRa technology in human germ cells (testicular seminoma cells—TCam-2). Successful activation of the ESX1 gene in TCam-2 cells using the CRISPRa system was achieved, and the expression level of the ESX1 gene was significantly higher in modified TCam-2 cells than in WT cells or the negative control with nontargeted gRNA (p < 0.01). Using RNA-seq, a network of over 50 genes potentially regulated by the ESX1 gene was determined. Finally, 6 genes, NANOG, CXCR4, RPS6KA5, CCND1, PDE1C, and LINC00662, participating in cell proliferation and differentiation were verified in azoospermic patients with and without a mutation in the ESX1 gene as well as in men with normal spermatogenesis, where inverse correlations in the expression levels of the observed genes were noted. |
| first_indexed | 2024-03-09T15:21:17Z |
| format | Article |
| id | doaj.art-3d4d30706a7146169ea59dfc53471e2b |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-03-09T15:21:17Z |
| publishDate | 2023-10-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-3d4d30706a7146169ea59dfc53471e2b2023-11-26T12:47:54ZengNature PortfolioScientific Reports2045-23222023-10-0113111410.1038/s41598-023-43854-9ESX1 gene as a potential candidate responsible for male infertility in nonobstructive azoospermiaAgnieszka Malcher0Zuzanna Graczyk1Hermann Bauer2Tomasz Stokowy3Andrea Berman4Mikołaj Smolibowski5Dominika Blaszczyk6Piotr Jedrzejczak7Alexander N. Yatsenko8Maciej Kurpisz9Institute of Human Genetics, Polish Academy of SciencesInstitute of Human Genetics, Polish Academy of SciencesDepartment of Developmental Genetics, Max Planck Institute for Molecular GeneticsIT Division, University of BergenDepartment of Biological Sciences, University of PittsburghInstitute of Human Genetics, Polish Academy of SciencesInstitute of Human Genetics, Polish Academy of SciencesDepartment of Cell Biology, Center of Obstetrics, Gynecology and Infertility Treatment, University of Medical SciencesDepartment of OB/GYN and Reproductive Sciences, School of Medicine, University of PittsburghInstitute of Human Genetics, Polish Academy of SciencesAbstract Infertility is a problem that affects approximately 15% of couples, and male infertility is responsible for 40–50% of these cases. The cause of male infertility is still poorly diagnosed and treated. One of the prominent causes of male infertility is disturbed spermatogenesis, which can lead to nonobstructive azoospermia (NOA). Whole-genome sequencing (WGS) allows us to identify novel rare variants in potentially NOA-associated genes, among others, in the ESX1 gene. The aim of this study was to activate the ESX1 gene using CRISPRa technology in human germ cells (testicular seminoma cells—TCam-2). Successful activation of the ESX1 gene in TCam-2 cells using the CRISPRa system was achieved, and the expression level of the ESX1 gene was significantly higher in modified TCam-2 cells than in WT cells or the negative control with nontargeted gRNA (p < 0.01). Using RNA-seq, a network of over 50 genes potentially regulated by the ESX1 gene was determined. Finally, 6 genes, NANOG, CXCR4, RPS6KA5, CCND1, PDE1C, and LINC00662, participating in cell proliferation and differentiation were verified in azoospermic patients with and without a mutation in the ESX1 gene as well as in men with normal spermatogenesis, where inverse correlations in the expression levels of the observed genes were noted.https://doi.org/10.1038/s41598-023-43854-9 |
| spellingShingle | Agnieszka Malcher Zuzanna Graczyk Hermann Bauer Tomasz Stokowy Andrea Berman Mikołaj Smolibowski Dominika Blaszczyk Piotr Jedrzejczak Alexander N. Yatsenko Maciej Kurpisz ESX1 gene as a potential candidate responsible for male infertility in nonobstructive azoospermia Scientific Reports |
| title | ESX1 gene as a potential candidate responsible for male infertility in nonobstructive azoospermia |
| title_full | ESX1 gene as a potential candidate responsible for male infertility in nonobstructive azoospermia |
| title_fullStr | ESX1 gene as a potential candidate responsible for male infertility in nonobstructive azoospermia |
| title_full_unstemmed | ESX1 gene as a potential candidate responsible for male infertility in nonobstructive azoospermia |
| title_short | ESX1 gene as a potential candidate responsible for male infertility in nonobstructive azoospermia |
| title_sort | esx1 gene as a potential candidate responsible for male infertility in nonobstructive azoospermia |
| url | https://doi.org/10.1038/s41598-023-43854-9 |
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