La valutazione della frammentazione del DNA spermatico nei soggetti infertili
The laboratory assessment of sperm DNA fragmentation in infertile patients

AUTORI

ABSTRACT

The laboratory assessment of sperm DNA fragmentation in infertile patients

Over 15% of couples worldwide suffer from infertility and in 50% of cases a male factor is found. According to the World Health Organization, sperm analysis is the most appropriate test to assess male infertility. Since quite often, the conventional semen parameters are related to sperm DNA damage, the integration of this evaluation with sperm DNA fragmentation (SDF) could independently predict the sperm reproductive potential. Unfortunately, this analysis has not entered into routine clinical practice. The aim of this review is to discuss the importance of the SDF analysis and its clinical implication and to evaluate the extrinsic and intrinsic factors that affect the DNA fragmentation. In addition, principles and protocols of different methods used to evaluate and quantify the SDF are reviewed; advantages and disadvantages of different methods are reported.

BIBLIOGRAFIA

1. Kumar N, Singh AK. Trends of male factor infertility, an important cause of infertility: A review of literature. J Hum Reprod Sci 2015;8:191-6.
2. Cariati F, D’Argenio V, Tomaiuolo R. The evolving role of genetic tests in reproductive medicine. J Transl Med 2019;17:267.
3. Cariati F, D’Uonno N, Borrillo F, et al. Bisphenol A: an emerging threat to male fertility. Reprod Biol Endocrinol 2019;17:6
4. Guzick DS, Overstreet JW, Factor-Litvak P, et al. Sperm morphology, motility, and concentration in fertile and infertile men. N Engl J Med 2001;345:1388-93.
5 Barratt CL, Mansell S, Beaton C, et al. Diagnostic tools in male infertility-the question of sperm dysfunction. Asian J Androl 2011;13:53-8.
6. Chen X, Zhang W, LuoY, et al. Predictive value of semen parameters in vitro fertilisation pregnancy outcome. Andrologia 2009;41:111-7.
7. Aitken RJ. Whither must spermatozoa wander? The future of laboratory seminology. Asian J Androl 2010;12:99-103.
8. Champroux A, Torres Carreira J, Gharagozloo P, et al. Mammalian sperm nuclear organization: resiliencies and vulnerabilities. Basic Clin Androl 2016;26:7.
9. Cariati F, Jaroudi S, Alfarawati S, et al. Investigation of sperm telomere length as a potential marker of paternal genome integrity and semen quality. Reprod BioMed Online 2016;33:404-11.
10. Zini A, Bielecki R, Phang D et al. Correlations between two markers of sperm DNA integrity, DNA denaturation and DNA fragmentation, in fertile and infertile men. Fertil Steril 2001;75:674-7.
11. Sakkas D, Alvarez JG. Sperm DNA fragmentation: mechanisms of origin, impact on reproductive outcome, and analysis. Fertil Steril. 2010;93:1027-36.
12. Sergerie M, Laforest G, Bujan L, et al. Sperm DNA fragmentation: threshold value in male fertility. Hum Reprod 2005;20:3446-51.
13. Osman A, Alsomait H, Seshadri S, et al. The effect of sperm DNA fragmentation on live birth rate after IVF or ICSI: a systematic review and meta-analysis. Reprod Biomed Online 2015;30:120-7.
14. Simon L, Zini A, Dyachenko A. A systematic review and meta-analysis to determine the effect of sperm DNA damage on in vitro fertilization and intracytoplasmic sperm injection outcome. Asian J Androl 2017;19:80-90.
15. Evenson DP, Wixon R. Data analysis of two in vivo fertility studies using Sperm Chromatin Structure Assay-derived DNA fragmentation index vs. pregnancy outcome. Fertil Steril 2008;90:1229-31.
16. D’Argenio V, Borrillo F, Cariati F, et al. Glossary of molecular biology and clinical molecular biology. Part I: general terms. Biochim Clin 2019,43:90-105.
17. Huckins C. The morphology and kinetics of spermatogonial degeneration in normal adult rats: an analysis using a simplified classification of the germinal epithelium. Anat Rec 1978;190:905-26.
18. Rodriguez I, Ody C, Araki K, et al. An early and massive wave of germinal cell apoptosis is required for the development of functional spermatogenesis. EMBO J 1997;16:2262-70.
19. Sakkas D, Mariethoz E, St John JC. Abnormal sperm parameters in humans are indicative of an abortive apoptotic mechanism linked to the Fas-mediated pathway. Exp Cell Res 1999;251:350-5.
20. De Lamirande E, Gagnon C. Capacitation-associated production of superoxide anion by human spermatozoa. Free Radic Biol Med 1995;18:487-95.
21. Gong S, San Gabriel MC, Zini A, et al. Low amounts and high thiol oxidation of peroxiredoxins in spermatozoa from infertile men. J Androl 2012;33:1342-51.
22. Simon L, Castillo J, Oliva R, et al. Relationships between human sperm protamines, DNA damage and assisted reproduction outcomes. Reprod Biomed Online 2011;23:724-34.
23. De Iuliis GN, Thomson LK, Mitchell LA, et al. DNA damage in human spermatozoa is highly correlated with the efficiency of chromatin remodeling and the formation of 8- hydroxy-2′-deoxyguanosine, a marker of oxidative stress. Biol Reprod 2009;81:517-24.
24. De Lamirande E, Gagnon C. Capacitation-associated production of superoxide anion by human spermatozoa. Free Radic Biol Med 1995;18:487-95.
25. Potts RJ, Jefferies TM, Notarianni LJ. Antioxidant capacity of the epididymis. Hum Reprod 1999;14:2513-6.
26. Agarwal A, Virk G, Ong C, et al. Effect of oxidative stress on male reproduction. World J Mens Health 2014;32:1-17.
27. Koppers AJ, De Iuliis GN, Finnie JM, et al. Significance of mitochondrial reactive oxygen species in the generation of oxidative stress in spermatozoa. J Clin Endocrinol Metab 2008;93:3199-207.
28. Lobascio AM, De Felici M, Anibaldi M, et al. Involvement of seminal leukocytes, reactive oxygen species, and sperm mitochondrial membrane potential in the DNA damage of the human spermatozoa. Andrology 2015;3:265-70.
29. Koppers AJ, Mitchell LA, Wang P, et al. Phosphoinositide 3- kinase signalling pathway involvement in a truncated apoptotic cascade associated with motility loss and oxidative DNA damage in human spermatozoa. Biochem J 2011;436:687-98.
30. Aitken RJ, Jones KT, Robertson SA. Reactive oxygen species and sperm function- -in sickness and in health. J Androl 2012;33:1096-106.
31. Bergamo P, Volpe MG, Lorenzetti S, et al. Human semen as an early, sensitive biomarker of highly polluted living environment in healthy men: A pilot biomonitoring study on trace elements in blood and semen and their relationship with sperm quality and RedOx status. Reprod Toxicol 2016;66:1-9.
32. Alviggi C, Cariati F, Conforti A et al. The effect of FT500 Plus® on ovarian stimulation in PCOS women. Reprod Toxicol 2016;59:40-4.
33. Aitken RJ, Koopman P, Lewis SE. Seeds of concern. Nature 2004;432:48-52.
34. Noblanc A, Damon Soubeyrand C, Karrich B, et al. DNA oxidative damage in mammalian spermatozoa: where and why is the male nucleus affected? Free Radic Biol Med 2003;65:719-23.
35. Bosco L, Notari T, Ruvolo G et al. Sperm DNA fragmentation: An early and reliable marker of air pollution. Environ Toxicol Pharmacol 2018;58:243-9.
36. Radwan M, Jurewicz J, Merecz-kot D, et al. Sperm DNA damage-the effect of stress and everyday life factors. Int J Impot Res 2016;28:148-54.
37. Sen S, Chakraborty R. The role of antioxidants in human health. In: Oxidative stress: diagnostics, prevention, and therapy. ACS Symposium Series 2011:1-37.
38. Talebi AR, Sarcheshmeh AA, Khalili MA, et al. Effects of ethanol consumption on chromatin condensation and DNA integrity of epididymal spermatozoa in rat. Alcohol 2011;45:403-9.
39. Azam S, Hadi N, Khan NU, et al. Antioxidant and prooxidant properties of caffeine, theobromine and xanthine. Med Sci Monitor 2003;9:325-30.
40. Hamad M, Shelko N, Kartarius S, et al. Impact of cigarette smoking on histone (H2B) to protamine ratio in human spermatozoa and its relation to sperm parameters.
Andrology 2014;2:666-77.
41. Boggia B, Carbone U, Farinaro E, et al. Effects of working posture and exposure to traffic pollutants on sperm quality. J Endocrinol Invest 2009;32:430-4.
42. Gallegos G, Ramos B, Santiso R, et al. Sperm DNA fragmentation in infertile men with genitourinary infection by Chlamydia trachomatis and Mycoplasma. Fertil Steril 2008;90:328-34.
43. Boeri L, Capogrosso P, Ventimiglia E, et al. High-risk human papillomavirus in semen is associated with poor sperm progressive motility and a high sperm DNA fragmentation index in infertile men. Hum Reprod 2019;34:209-17.
44. Garolla A, Engl B, Pizzol D, et al. Spontaneous fertility and in vitro fertilization outcome: new evidence of human papillomavirus sperm infection. Fertil Steril 2016;105:65-72.
45. Garolla A, De Toni L, Bottacin A, et al. Human Papillomavirus Prophylactic Vaccination improves reproductive outcome in infertile patients with HPV semen infection: a retrospective study. Sci Rep 2018;8:912.
46. Agbaje I, Rogers D, McVicar C, et al. Insulin dependent diabetes mellitus: implications for male reproductive function. Hum Reprod 2007;22:1871-7.
47. Maselli J, Hales BF, Chan P, Robaire B. Exposure to bleomycin, etoposide, and cis-platinum alters rat sperm chromatin integrity and sperm head protein profile. Biol Reprod 2012;86:166.
48. Smith R, Kaune H, Parodi D, et al. Increased sperm DNA damage in patients with varicocele: relationship with seminal oxidative stress. Hum Reprod 2006;21:986-93.
49. Gorczyca W, Traganos F, Jesionowska H, et al. Presence of DNA strand breaks and increased sensitivity of DNA in situ to denaturation in abnormal human sperm cells: analogy to apoptosis of somatic cells. Exp Cell Res 1993;207:202-5.
50. Gold R, Schmied M, Rothe G et al. Detection of DNA fragmentation in apoptosis: application of in situ nick translation to cell culture systems and tissue sections. J Histochem Cytochem 1993;41:1023-30.
51. Hughes C, Lewis S, McKelvey-Martin V, Thompson W. A comparison of baseline and in- duced DNA damage in human spermatozoa from fertile and infertile men, using a modified comet assay. Mol Hum Reprod 1996;2:613-9.
52. Fernandez JL, Vazquez-Gundin F, Delgado A, et al. DNA breakage detection-FISH (DBD-FISH) in human spermatozoa: technical variants evidence different structural features. Mutat Res 2000;453:77-82 .
53. Evenson DP, Larson KL, Jost LK. Sperm chromatin structure assay: its clinical use for detecting sperm DNA fragmentation in male infertility and comparisons with other techniques. J Androl 2002;23:25-43.
54. Fernandez JL, Muriel L, Rivero MT, et al. The sperm chromatin dispersion test: a simple method for the determination of sperm DNA fragmentation. J Androl 2003;24:59-66.
55. Sharma R, Ahmad G, Esteves SC, et al. Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay using bench top flow cytometer for evaluation of sperm DNA fragmentation in fertility laboratories: protocol, reference values, and quality control. J Assist Reprod Genet 2016;33:291-300.
56. Di Maggio F, Borrillo F, Cariati F, et al. Glossario di biologia molecolare e biologia molecolare clinica. Parte II: metodologie di biologia molecolare. Biochim Clin
2019;43:435-48.
57. Sergerie M, Laforest G, Bujan L, et al. Sperm DNA fragmentation: threshold value in male fertility. Hum Reprod 2005;20:3446-51.
58. Sun JG, Jurisicova A, Casper RF. Detection of deoxyribonucleic acid fragmentation in human sperm: correlation with fertilization in vitro. Biol Reprod 1997;56:602-7.
59. Duran EH, Morshedi M, Taylor S, et al. Sperm DNA quality predicts intrauterine insemination outcome: a prospective cohort study. Hum Reprod 2002;17:3122-8.
60. Borini A, Tarozzi N, Bizzaro D, et al. Sperm DNA fragmentation: paternal effect on early post-implantation embryo development in ART. Hum Reprod 2006;21:2876-81.
61. Tarozzi N, Bizzaro D, Flamigni C, et al. Clinical relevance of sperm DNA damage in assisted reproduction. Reprod Biomed Online 2007;14:746-57.
62. Luchetti F, Canonico B, Biagiarelli L, et al. Indagini citofluorimetriche nella vitalità e morte cellulare. I. Necrosi, apoptosi e proliferazione cellulare. Biochim Clin, 2009,33:83-92.
63. Evenson DP, Larson KL, Jost LK. Sperm chromatin structure assay: its clinical use for detecting sperm DNA fragmentation in male infertility and comparisons with other techniques. J Androl 2002;23:25-43.
64. Fraser L. Structural damage to nuclear DNA in mammalian spermatozoa: its evaluation techniques and relationship with male infertility. Pol J Vet Sci 2004;7:311-21.
65. Spano M, Kolstad AH, Larsen SB, et al. The applicability of the flow cytometric sperm chromatin structure assay in epidemiological studies. Asclepios Hum Reprod 1998;13:2495-505.
66. Evgeni E, Charalabopoulos K, Asimakopoulos B. Human sperm DNA fragmentation and its correlation with conventional semen parameters. J Reprod Infertil 2014;15:2-14.
67. Boe-Hansen GB, Ersboll AK, Christensen P. Variability and laboratory factors affecting the sperm chromatin structure assay in human semen. J Androl 2005;26:360-8.
68. Ferna ́ ndez JL, Muriel L, Goyanes V, et al. Simple determination of human sperm DNA fragmentation with an improved sperm chromatin dispersion test. Fertil Steril 2005;84:833-42.
69. Manesh KPS, Ashok A. A systematic review on sperm DNA fragmentation in male factor infertility: Laboratory assessment. Arab J Urol 2018;16:65-76.
70. Simon L, Murphy K, Shamsi MB et al. Paternal influence of sperm DNA integrity on early embryonic development. Hum Reprod 2014;29:2402-12.
71. Donnelly ET, Steele EK, McClure N et al. Assessment of DNA integrity and morphology of ejaculated spermatozoa from fertile and infertile men before and after cryopreservation. Hum Reprod 2001;16:1191-9.
72. Singh NP, McCoy MT, Tice RR et al. A simple technique for quantitation of low levels of DNA damage in individual cells. Exp Cell Res 1988;175:184-91.
73. Ferna ndez JL, Muriel L, Rivero MT, et al. The sperm chromatin dispersion test: a simple method for the determination of sperm DNA fragmentation. J Androl 2003;24:59-66.
74. Bungum M, Humaidan P, Axmon A, et al. Sperm DNA integrity assessment in prediction of assisted reproduction technology outcome. Hum Reprod 2007;22:174-9.
75. Sá R, Cunha M, Rocha E, et al. Sperm DNA fragmentation is related to sperm morphological staining patterns. Reprod Biomed Online 2015;31:506-15.
76. Tomaiuolo R, Panteghini M. Il laboratorio nella medicina della riproduzione. Biochim Clin, 2017;41:292-3.
77. Cariati F, Savarese M, D’Argenio V, et al. The SEeMORE strategy: Single-tube electrophoresis analysis-based genotyping to detect monogenic diseases rapidly and effectively from conception until birth. Clin Chem Lab Med 2017;56:40-50.
78. Tomaiuolo R, Fausto M, Elce A, et al. Enhanced frequency of CFTR gene variants in couples who are candidates for assisted reproductive technology treatment. Clin Chem
Lab Med 2011;49:1289-93.
79. D’Argenio V, Nunziato M, D’Uonno N, et al. Indicazioni e limiti della diagnosi genetica preimpianto. Biochim Clin 2017;41:314-21.
80. Zini A, Sigman M. Are tests of sperm DNA damage clinically useful? Pros and cons. J Androl 2009;30:219-29.
81. Khadem N, Poorhoseyni A, Jalali M, et al. Sperm DNA fragmentation in couples with unexplained recurrent spontaneous abortions. Andrologia 2014;46:126-30.
82. Gosálvez C, Lopez Fernandez C, Fernandez JL, et al. Unpacking the mysteries of sperm DNA fragmentation:
Ten frequently asked questions. J Reprod Biotechnol Fertil 2015;4:1-16.
83. Sivanarayana T, Rava Krishna T, Jaya Prakah G, et al. Sperm DNA fragmentation assay by sperm chromatin dispersion (SCD): correlation between DNA fragmentation and outcome of intracytoplasmic sperm injection. RMB 2014;13:87-94.
84. De Angelis C, Galdiero M, Pivonello C, et al. The role of vitamin D in male fertility: A focus on the testis. Rev Endocr Metab Disord 2017;18:285.
85. Pacey AA. Environmental and lifestyle factors associated with sperm DNA damage. Hum Fertil 2010;13:189-93.