Андрология и генитальная хирургия. 2024; 25: 64-75
Нарушение сперматогенеза и мейоза у пациентов с мозаицизмом и/или структурными аномалиями Y-хромосомы
https://doi.org/10.62968/2070-9781-2024-25-4-64-76Аннотация
Введение. Структурные несбалансированные аномалии и патогенные микроделеции Y-хромосомы являются частой генетической причиной мужского бесплодия вследствие нарушения сперматогенеза. Аномалии половых хромосом (гоносом) часто связаны с гоносомным мозаицизмом, наличие, тип и выраженность которого могут модифицировать негативный эффект анеусомий, структурных вариантов половых хромосом, Х- и Y-сцепленных мутаций. Недостаточно изучен совместный патогенный эффект структурных вариантов и мозаицизма Y-хромосомы на сперматогенез, мейоз и мужскую фертильность.
Цель работы. Исследование генетических и спермиологических нарушений у пациентов с мозаицизмом и/или структурными аномалиями Y-хромосомы.
Материалы и методы. Обследована группа из 40 мужчин возраста 31±7 (от 16 до 49) лет, у которых в кариотипе обнаружены мозаицизм и/или структурные аномалии/варианты Y-хромосомы. Кариотип определяли по результатам стандартного цитогенетического исследования (на культивированных лимфоцитах периферической крови), у части пациентов дополнительно выполняли FISH исследование с ДНК-зондами для хромосом X и Y. Наличие Y-специфичных последовательностей локусов SRY и AZF(a,b,c) определяли методом полимеразной цепной реакции (ПЦР). Стандартное спермиологическое исследование выполняли согласно рекомендациям лабораторного Руководства ВОЗ (2010), у 12 мужчин выполнен количественный кариологический анализ (ККА) незрелых половых клеток (НПК).
Результаты. У 29 (72,5%) мужчин выявлен мозаицизм по хромосоме Y.Патогенные делеции в локусе AZF (AZFa+b+c, n=3; AZFb+c, n=19; AZFc, n=3) детектированы у 25 (62,5%) пациентов, из них у 17 пациентов обнаружен мозаицизм. Необструктивная азооспермия и тяжелая олигозооспермия диагностированы у 88% и 12% мужчин с делециями AZF, и у 47% и 20% пациентов без AZF делеций, соответственно. Нормозооспермия и нетяжелые формы патозооспермии выявлены только среди мужчин без патогенных делеций в локусе AZF. У пациентов с кольцевыми и псевдодицентрическими хромосомами Y, терминальными делециями Yq11.2 обнаружены только тяжелые формы нарушения сперматогенеза. ККА НПК выявил криптозооспермию у 7 из 10 обследованных пациентов с азооспермией (по данным спермограммы), а также признаки нарушения деления сперматоцитов в пахитене и/или на допахитенных стадиях профазы I мейоза.
Заключение. Сперматологические нарушения у мужчин с мозаицизмом и/или структурными аномалиями хромосомы Y выраженно варьируют от их отсутствия до тяжелых форм. Характер и тяжесть поражения сперматогенеза зависит от варианта структурной аномалии, типа (и генного состава) CNV, локализации точек разрыва, дополнительных патогенных факторов. Гоносомный мозаицизм является дополнительным патогенным нарушающим фертильность фактором, который может утяжелять генетические и фенотипические изменения.
Список литературы
1. Garcia-Herrero S, Simon B, Garcia-Planells J. The Reproductive Journey in the Genomic Era: From Preconception to Childhood. Genes (Basel). 2020 Dec 19;11(12):1521. doi: 10.3390/genes11121521. PMID: 33352697; PMCID: PMC7767043
2. Черных В.Б. Макро- и микроструктурные перестройки Y-хромосомы. Медицинская генетика. 2007. – №10. – С. 45-52.
3. Hsu LY. Phenotype/karyotype correlations of Y chromosome aneuploidy with emphasis on structural aberrations in postnatally diagnosed cases. Am J Med Genet. 1994;53(2):108-40. doi: 10.1002/ajmg.1320530204
4. Krausz C, Casamonti E. Spermatogenic failure and the Y chromosome. Hum. Genet. 2017, 136, 637–655. doi: 10.1007/s00439-017-1793-8
5. Черных В.Б. Гоносомные аномалии и CNV и их диагностика. Медицинская генетика. 2018;17(10):8-14. https://doi.org/10.25557/2073-7998.2018.10.8-14
6. Черных В.Б. AZF делеции – частая генетическая причина бесплодия у мужчин: современное состояние исследований. Пробл. репрод. 2009;15(1):10-15.
7. Patsalis PC, Skordis N, Sismani C et al. Identification of high frequency of Y chromosome deletions in patients with sex chromosome mosaicism and correlation with the clinical phenotype and Y-chromosome instability. Am J Med Genet A. 2005 Jun 1;135(2):145-9. doi: 10.1002/ajmg.a.30712. PMID: 15880425.
8. McGovan-Jordan J., Hastings R.J., Moore S. ISCN 2020 – An International System for Human Cytogenetic Nomenclature (2020) Karger, 2020: 170р.
9. Hsieh CL. Basic cytogenetic techniques: culturing, slide making, and G-banding. In Cell Biology: A Laboratory Handbook, (2nd edition.), 1998, Vol.3. Elsevier, Academic Press. New York.
10. Bates S.E. Classical cytogenetics: karyotyping techniques Methods Mol. Biol., 767 (2011), pp. 177-190
11. Gu J, Smith JL, Dowling PK. Fluorescence In Situ Hybridization Probe Validation for Clinical Use. Methods Mol Biol. 2017;1541:101-118. doi: 10.1007/978-1-4939-6703-2_10. PMID: 27910018.
12. Romczak EZ, Punnett HH. An improved method for chromosome enumeration using fluorescent in situ hybridization of buccal epithelium. J Ass Genet Tech. 1998;24:6-7.
13. Moore LE, Titenko-Holland N, Smith MT. Use of fluorescence in situ hybridization to detect chromosome-specific changes in exfoliated human bladder and oral mucosa cells. Environ Mol Mutagen. 1993;22(3):130-7. doi: 10.1002/em.2850220304. PMID: 8404872.
14. Paulo Jd, Andrade JG, Santos AP et al. The use of FISH on buccal smear to investigate mosaicism with a 45,X cell line: study on healthy men and patients with disorders of sex development. Arq Bras Endocrinol Metabol. 2014 Jun;58(4):328- 34. Portuguese. doi: 10.1590/0004-2730000002853. PMID: 24936726.
15. Krausz C, Navarro-Costa P, Wilke M et al. EAA/EMQN best practice guidelines for molecular diagnosis of Y-chromosomal microdeletions: State of the art 2023. Andrology. 2023 Sep 6. doi: 10.1111/andr.13514.
16. Черных В.Б., Рыжкова О.П., Кузнецова И.А. и др. Делеции AZFс региона Y-хромосомы у российских фертильных мужчин. Генетика, 2022, T. 58, № 7, С. 838-845.
17. Руководство ВОЗ по исследованию и обработке эякулята человека. Пер. с англ. Н.П. Макарова. Науч. ред. Л.Ф. Курило. 5-е изд. М.: Капитал Принт, 2012. 291 с. World Health Organization. WHO Laboratory Manual for the Examination and Processing of Human Semen, 5th ed. Geneva: World Health Organization; 2010, 287 pp.
18. Курило Л.Ф., Дубинская В.П., Остроумова Т.В. и др. Оценка сперматогенеза по незрелым половым клеткам эякулята. Проблемы репродукции. 1995; 3: 33-38.
19. Chang HJ, Clark RD, Bachman H. The phenotype of 45,X/46,XY mosaicism: an analysis of 92 prenatally diagnosed cases. Am J Hum Genet. 1990;46(1):156-167.
20. Mohammadpour Lashkari F, Sadighi Gilani MA, Ghaheri A et al. Clinical aspects of 49 infertile males with 45,X/46,XY mosaicism karyotype: A case series // Andrologia. 2018; e13009. https://doi.org/10.1111/and.13009
21. Li L, Zhang H, Yang Y et al. High frequency of Y chromosome microdeletions in male infertility patients with 45,X/46,XY mosaicism. Braz J Med Biol Res. 2020 Feb 14;53(3):e8980. doi: 10.1590/1414-431X20198980. eCollection 2020.
22. Valetto A, Bertini V, Rapalini E et al. Molecular and cytogenetic characterization of a structural rearrangement of the Y chromosome in an azoospermic man. Fertil Steril 2004; 81: 1388- 1390. doi: 10.1016/j.fertnstert.2003.09.069
23. Siffroi JP, Le Bourhis C, Krausz C et al. Sex chromosome mosaicism in males carrying Y chromosome long arm deletions. Hum Reprod. 2000; 15(12): 2559-2562. doi: 10.1093/humrep/15.12.2559.
24. Wu Q, Wang C, Shi H et al. The clinical manifestation and genetic evaluation in patients with 45,X/46,XY Mosaicism. Sex Dev 2017; 11: 64-69. DOI: 10.1159/000455260
25. Le Bourhis C, Siffroi JP, McElreavey K et al. Y chromosome microdeletions and germinal mosaicism in infertile males. Mol Hum Reprod. 2000;6(8):688-693. doi: 10.1093/molehr/6.8.688.
26. Hong-Ge Li, Li-Hong Fan, Bei Liu et al. The association between the two more common genetic causes of spermatogenic failure: a 7-year retrospective study. Asian Journal of Andrology 2020; 22, 642–648; doi: 10.4103/aja.aja_13_20
27. Wu YH, Sun KN, Bao H et al. SRY-negative 45,X/46,XY adult male with complete masculinization and infertility: A case report and review of literature. World J Clin Cases. 2020;8(24):6380-6388. doi: 10.12998/wjcc.v8.i24.6380.
28. Rosa RF, D’Ecclesiis WF, Dibbi RP et al. 45,X/46,XY mosaicism: report on 14 patients from a Brazilian hospital. A retrospective study. Sao Paulo Med J. 2014;132(6):332-338. doi: 10.1590/1516-3180-2014-1326729.
29. Ketheeswaran S, Alsbjerg B, Christensen P et al. 45,X/46,XY mosaicism and normozoospermia in a patient with male phenotype. Case Rep Med. 2019;2019:2529080. doi: 10.1155/2019/2529080.
30. Valetto A, Bertini V, Rapalini E et al. Molecular and cytogenetic characterization of a structural rearrangement of the Y chromosome in an azoospermic man. Fertil Steril 2004; 81: 1388- 1390. DOI: 10.1016/j.fertnstert.2003.09.069
31. Lehmann KJ, Kovac JR, Xu J et al. Isodicentric Yq mosaicism presenting as infertility and maturation arrest without altered SRY and AZF regions. J Assist Reprod Genet. 2012;29(9):939-42. doi: 10.1007/s10815-012-9822-y.
32. Lindhardt Johansen M, Hagen CP, Rajpert-De Meyts E. et al. 45,X/46,XY mosaicism: phenotypic characteristics, growth, and reproductive function – a retrospective longitudinal study. J Clin Endocrinol Metab. 2012;97(8):E1540-9. doi: 10.1210/jc.2012-1388.
33. Vogt PH, Edelmann A, Kirsch S et al. Human Y chromosome azoospermia factors (AZF) mapped to different subregions in Yq11. Hum Mol Genet. 1996; 5(7): 933-943. doi: 10.1093/hmg/5.7.933.
34. Gardner RJ, McKinlay Amor DJ. Gardner and Sutherland’s Chromosome Abnormalities and Genetic Counseling. Oxford University Press. Hum Genet 137, 971 (2018) https://doi.org/10.1007/s00439-018-1946-4.
35. Опарина Н.В., Райгородская Н.Ю., Латышев О.Ю. и др. Тканевой гоносомный мозаицизм у пациентов с нарушением формирования пола, связанным с аномалиями дифференцировки гонад. Генетика, 2021, T. 57 (11). С. 1306-1317. doi: 10.31857/S0016675821110102
36. Guo X, Dai X, Zhou T et al. Mosaic loss of human Y chromosome: what, how and why. Hum Genet. 2020 Apr;139(4):421-446. doi: 10.1007/s00439-020-02114-w.
Andrology and Genital Surgery. 2024; 25: 64-75
Spermatogenesis and meiosis defects in patients with mosaicism and/or the Y chromosome structural abnormalities
https://doi.org/10.62968/2070-9781-2024-25-4-64-76Abstract
Unbalanced structural Y chromosome abnormalities and pathogenic Y chromosome microdeletions are common genetic cause of severe male infertility due to spermatogenesis damage. Sex chromosomes (gonosomes) abnormalities are often associated with mosaicism the presence and percentage of gonosomal mosaicism may modify the damage of the negative effect of sex chromosome abnormalities and X- and Y-linked mutations. The combined effect of structural abnormalities and mosaicism of the Y chromosome on male fertility and semen parameters has not been sufficiently studied. The aim of the study was the analysis of the spermatological and genetic examination in mosaic patients with/or without structural abnormalities of the Y chromosome to evaluate the spermatogenesis and meiosis defects.
Material and methods. A group of 40 men aged 31±7 (from 16 to 49) years was examined, mosaicism and/or structural abnormalities/variants of the Y chromosome were found in the karyotype of patients. The karyotype detected on the results of a standard cytogenetic study (on cultured peripheral blood lymphocytes), a FISH study with DNA probes for X and Y chromosomes was additionally performed. The presence/absence of Y-specific sequences (SRY and AZFa,b,c loci) was determined by multiplex polymerase chain reaction. A standard spermiological examination was performed according to the WHO Laboratory Manual, quantitative karyological analysis (QKA) of immature germ cells (IGCs) was performed in 12 patients.
Results. Y chromosome mosaicism was found in 29 (72.5%) patients. Pathogenic AZF microdeletions (AZFa+b+c, n=3; AZFb+c, n=19; AZFc, n=3) were detected in 25 (62.5%) men, 17 of them had the Y chromosome mosaicism. Non-obstructive azoospermia and severe oligozoospermia were found in 88% and 12% of men with AZF deletions, and without AZF deletions in 47% and 20% of patients, respectively. Normozoospermia and mild forms of pathozoospermia were revealed in patients without AZF deletions. Only severe forms of spermatogenesis defects were found in all men with Yq11.2 deletions, ring and pseudodicentric Y chromosomes. QKA of IGCs revealed cryptozoospermia in 7 out of 10 azoospermic patients (according to semen analyses data) and signs of a partial meiotic arrest of spermatogenesis at pachytene and/or at the pre-pachytene stages of prophase I of meiosis.
Conclusion. In patients with mosaicism and/or structural abnormalities of the Y chromosome, the spermatogenesis disorders vary from their absence to their severe forms. The severity of the spermatogenesis depletion depends on structural anomalies and Yq11.2(AZF) deletions, their type, and the breakpoints location, the additional pathogenic factors. Gonosomal mosaicism is an additional pathogenic factor affected fertility that can increase genetic and phenotypic changes.
References
1. Garcia-Herrero S, Simon B, Garcia-Planells J. The Reproductive Journey in the Genomic Era: From Preconception to Childhood. Genes (Basel). 2020 Dec 19;11(12):1521. doi: 10.3390/genes11121521. PMID: 33352697; PMCID: PMC7767043
2. Chernykh V.B. Makro- i mikrostrukturnye perestroiki Y-khromosomy. Meditsinskaya genetika. 2007. – №10. – S. 45-52.
3. Hsu LY. Phenotype/karyotype correlations of Y chromosome aneuploidy with emphasis on structural aberrations in postnatally diagnosed cases. Am J Med Genet. 1994;53(2):108-40. doi: 10.1002/ajmg.1320530204
4. Krausz C, Casamonti E. Spermatogenic failure and the Y chromosome. Hum. Genet. 2017, 136, 637–655. doi: 10.1007/s00439-017-1793-8
5. Chernykh V.B. Gonosomnye anomalii i CNV i ikh diagnostika. Meditsinskaya genetika. 2018;17(10):8-14. https://doi.org/10.25557/2073-7998.2018.10.8-14
6. Chernykh V.B. AZF deletsii – chastaya geneticheskaya prichina besplodiya u muzhchin: sovremennoe sostoyanie issledovanii. Probl. reprod. 2009;15(1):10-15.
7. Patsalis PC, Skordis N, Sismani C et al. Identification of high frequency of Y chromosome deletions in patients with sex chromosome mosaicism and correlation with the clinical phenotype and Y-chromosome instability. Am J Med Genet A. 2005 Jun 1;135(2):145-9. doi: 10.1002/ajmg.a.30712. PMID: 15880425.
8. McGovan-Jordan J., Hastings R.J., Moore S. ISCN 2020 – An International System for Human Cytogenetic Nomenclature (2020) Karger, 2020: 170r.
9. Hsieh CL. Basic cytogenetic techniques: culturing, slide making, and G-banding. In Cell Biology: A Laboratory Handbook, (2nd edition.), 1998, Vol.3. Elsevier, Academic Press. New York.
10. Bates S.E. Classical cytogenetics: karyotyping techniques Methods Mol. Biol., 767 (2011), pp. 177-190
11. Gu J, Smith JL, Dowling PK. Fluorescence In Situ Hybridization Probe Validation for Clinical Use. Methods Mol Biol. 2017;1541:101-118. doi: 10.1007/978-1-4939-6703-2_10. PMID: 27910018.
12. Romczak EZ, Punnett HH. An improved method for chromosome enumeration using fluorescent in situ hybridization of buccal epithelium. J Ass Genet Tech. 1998;24:6-7.
13. Moore LE, Titenko-Holland N, Smith MT. Use of fluorescence in situ hybridization to detect chromosome-specific changes in exfoliated human bladder and oral mucosa cells. Environ Mol Mutagen. 1993;22(3):130-7. doi: 10.1002/em.2850220304. PMID: 8404872.
14. Paulo Jd, Andrade JG, Santos AP et al. The use of FISH on buccal smear to investigate mosaicism with a 45,X cell line: study on healthy men and patients with disorders of sex development. Arq Bras Endocrinol Metabol. 2014 Jun;58(4):328- 34. Portuguese. doi: 10.1590/0004-2730000002853. PMID: 24936726.
15. Krausz C, Navarro-Costa P, Wilke M et al. EAA/EMQN best practice guidelines for molecular diagnosis of Y-chromosomal microdeletions: State of the art 2023. Andrology. 2023 Sep 6. doi: 10.1111/andr.13514.
16. Chernykh V.B., Ryzhkova O.P., Kuznetsova I.A. i dr. Deletsii AZFs regiona Y-khromosomy u rossiiskikh fertil'nykh muzhchin. Genetika, 2022, T. 58, № 7, S. 838-845.
17. Rukovodstvo VOZ po issledovaniyu i obrabotke eyakulyata cheloveka. Per. s angl. N.P. Makarova. Nauch. red. L.F. Kurilo. 5-e izd. M.: Kapital Print, 2012. 291 s. World Health Organization. WHO Laboratory Manual for the Examination and Processing of Human Semen, 5th ed. Geneva: World Health Organization; 2010, 287 pp.
18. Kurilo L.F., Dubinskaya V.P., Ostroumova T.V. i dr. Otsenka spermatogeneza po nezrelym polovym kletkam eyakulyata. Problemy reproduktsii. 1995; 3: 33-38.
19. Chang HJ, Clark RD, Bachman H. The phenotype of 45,X/46,XY mosaicism: an analysis of 92 prenatally diagnosed cases. Am J Hum Genet. 1990;46(1):156-167.
20. Mohammadpour Lashkari F, Sadighi Gilani MA, Ghaheri A et al. Clinical aspects of 49 infertile males with 45,X/46,XY mosaicism karyotype: A case series // Andrologia. 2018; e13009. https://doi.org/10.1111/and.13009
21. Li L, Zhang H, Yang Y et al. High frequency of Y chromosome microdeletions in male infertility patients with 45,X/46,XY mosaicism. Braz J Med Biol Res. 2020 Feb 14;53(3):e8980. doi: 10.1590/1414-431X20198980. eCollection 2020.
22. Valetto A, Bertini V, Rapalini E et al. Molecular and cytogenetic characterization of a structural rearrangement of the Y chromosome in an azoospermic man. Fertil Steril 2004; 81: 1388- 1390. doi: 10.1016/j.fertnstert.2003.09.069
23. Siffroi JP, Le Bourhis C, Krausz C et al. Sex chromosome mosaicism in males carrying Y chromosome long arm deletions. Hum Reprod. 2000; 15(12): 2559-2562. doi: 10.1093/humrep/15.12.2559.
24. Wu Q, Wang C, Shi H et al. The clinical manifestation and genetic evaluation in patients with 45,X/46,XY Mosaicism. Sex Dev 2017; 11: 64-69. DOI: 10.1159/000455260
25. Le Bourhis C, Siffroi JP, McElreavey K et al. Y chromosome microdeletions and germinal mosaicism in infertile males. Mol Hum Reprod. 2000;6(8):688-693. doi: 10.1093/molehr/6.8.688.
26. Hong-Ge Li, Li-Hong Fan, Bei Liu et al. The association between the two more common genetic causes of spermatogenic failure: a 7-year retrospective study. Asian Journal of Andrology 2020; 22, 642–648; doi: 10.4103/aja.aja_13_20
27. Wu YH, Sun KN, Bao H et al. SRY-negative 45,X/46,XY adult male with complete masculinization and infertility: A case report and review of literature. World J Clin Cases. 2020;8(24):6380-6388. doi: 10.12998/wjcc.v8.i24.6380.
28. Rosa RF, D’Ecclesiis WF, Dibbi RP et al. 45,X/46,XY mosaicism: report on 14 patients from a Brazilian hospital. A retrospective study. Sao Paulo Med J. 2014;132(6):332-338. doi: 10.1590/1516-3180-2014-1326729.
29. Ketheeswaran S, Alsbjerg B, Christensen P et al. 45,X/46,XY mosaicism and normozoospermia in a patient with male phenotype. Case Rep Med. 2019;2019:2529080. doi: 10.1155/2019/2529080.
30. Valetto A, Bertini V, Rapalini E et al. Molecular and cytogenetic characterization of a structural rearrangement of the Y chromosome in an azoospermic man. Fertil Steril 2004; 81: 1388- 1390. DOI: 10.1016/j.fertnstert.2003.09.069
31. Lehmann KJ, Kovac JR, Xu J et al. Isodicentric Yq mosaicism presenting as infertility and maturation arrest without altered SRY and AZF regions. J Assist Reprod Genet. 2012;29(9):939-42. doi: 10.1007/s10815-012-9822-y.
32. Lindhardt Johansen M, Hagen CP, Rajpert-De Meyts E. et al. 45,X/46,XY mosaicism: phenotypic characteristics, growth, and reproductive function – a retrospective longitudinal study. J Clin Endocrinol Metab. 2012;97(8):E1540-9. doi: 10.1210/jc.2012-1388.
33. Vogt PH, Edelmann A, Kirsch S et al. Human Y chromosome azoospermia factors (AZF) mapped to different subregions in Yq11. Hum Mol Genet. 1996; 5(7): 933-943. doi: 10.1093/hmg/5.7.933.
34. Gardner RJ, McKinlay Amor DJ. Gardner and Sutherland’s Chromosome Abnormalities and Genetic Counseling. Oxford University Press. Hum Genet 137, 971 (2018) https://doi.org/10.1007/s00439-018-1946-4.
35. Oparina N.V., Raigorodskaya N.Yu., Latyshev O.Yu. i dr. Tkanevoi gonosomnyi mozaitsizm u patsientov s narusheniem formirovaniya pola, svyazannym s anomaliyami differentsirovki gonad. Genetika, 2021, T. 57 (11). S. 1306-1317. doi: 10.31857/S0016675821110102
36. Guo X, Dai X, Zhou T et al. Mosaic loss of human Y chromosome: what, how and why. Hum Genet. 2020 Apr;139(4):421-446. doi: 10.1007/s00439-020-02114-w.
