Frequency of Sperm Aneuploidy in Oligoasthenoteratozoospermic (OAT) Patients by Comprehensive Chromosome Screening: A Proof of Concept

Saei, Parishad; Bazrgar, Masood; Gourabi, Hamid; Kariminejad, Roxana; Eftekhari-Yazdi, Poopak; Fakhri, Mostafa

doi:10.18502/jri.v22i1.4996

Frequency of Sperm Aneuploidy in Oligoasthenoteratozoospermic (OAT) Patients by Comprehensive Chromosome Screening: A Proof of Concept

Authors

Parishad Saei ¹

Masood Bazrgar ²

Hamid Gourabi ²

Roxana Kariminejad ³

Poopak Eftekhari-Yazdi ⁴

Mostafa Fakhri ²

¹ Department of Genetics, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran; Department of Molecular Genetics, Faculty of Basic Sciences and Advanced Technologies in Biology,

² Department of Genetics, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran

³ Kariminejad-Najmabadi Pathology and Genetics Center, Tehran, Iran

⁴ Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran

10.18502/jri.v22i1.4996

Abstract

Background: Embryonic aneuploidy usually results in implantation failure and miscarriage. Considering significantly high frequency of sperm aneuploidy reported in oligoasthenoteratozoospermia (OAT) using fluorescence in situ hybridization (FISH) in limited number of chromosomes and lack of comprehensive chromosome screening (CCS) in OAT, the aim of this study was applying CCS in OAT sperm and comparison of the results with FISH findings. Methods: Five OAT patients with normal blood karyotypes and history of implantation failure were included. The successfully amplified samples, each containing two sperm, were analyzed by array comparative genomic hybridization (aCGH). FISH was utilized mainly depending on the aneuploidies found by aCGH to assess their frequencies in total sperm population. Results: In aCGH for 30 sperm, aneuploidy was found in 66% of samples. Following the study of 4300 sperm by FISH, an average of 55.46% aneuploidy was observed. No pregnancy was resulted with normal partners. Conclusion: Using aCGH, some abnormalities were observed that are not typically considered in sperm FISH studies. Despite small sample size of the comprehensive study, like other similar studies, the frequency of aneuploidies was considerable and similar to FISH. Aneuploidies revealed by aCGH at single sperm resolution were different from sperm population detected by FISH. Considering high frequency of aneuploidy in OATs sperm, preimplantation genetic testing for aneuploidy (PGT-A) can be used for in transfer of chromosomally normal embryos.

Keywords

Aneuploidy

Array comparative genomic hybridization

Fluorescence

In situ hybridization

Oligospermia

Spermatozoa

Singh R, Singh K. Male infertility: understanding, causes and treatment. 1st ed. Singapore: Springer; 2017. 497 p.

Martin RH, Ko E, Chan K. Detection of aneuploidy in human interphase spermatozoa by fluorescence in situ hybridization (FISH). Cytogenet Cell Genet. 1993;64(1):23-6.

Munné S, Chen S, Colls P, Garrisi J, Zheng X, Cekleniak N, et al, Maternal age, morphology, development and chromosome abnormalities in over 6000 cleavage-stage embryos. Reprod Biomed Online. 2007;14(5):628-34.

Templado C, Vidal F, Estop A. Aneuploidy in human spermatozoa. Cytogenet Genome Res. 2011;133(2-4):91-9.

Calogero AE, Burrello N, De Palma A, Barone N, D'Agata R, Vicari E. Sperm aneuploidy in infertile men. Reprod Biomed Online. 2003;6(3):310-7.

Piomboni, Paola, Anita Stendardi, and Laura Gambera. Chromosomal aberrations and aneuploidies of spermatozoa. In: Genetic damage in human spermatozoa. New York: Springer; 2014. p. 27-52.

Harton GL, Tempest HG. Chromosomal disorders and male infertility. Asian J Androl. 2012;14(1):32-9.

Rubio C, Gil-Salom M, Simon C, Vidal F, Rodrigo L, Minguez Y, et al. Incidence of sperm chromosomal abnormalities in a risk population: relationship with sperm quality and ICSI outcome. Hum Reprod. 2001;16(10):2084-92.

Pang MG, Hoegerman SF, Cuticchia AJ, Moon SY, Doncel GF, Acosta AA, et al. Detection of aneuploidy for chromosomes 4, 6, 7, 8, 9, 10, 11, 12, 13, 17, 18, 21, X and Y by fluorescence in-situ hybridization in spermatozoa from nine patients with oligoasthenoteratozoospermia undergoing intracytoplasmic sperm injection. Hum Reprod. 1999;14(5):1266-73.

Pfeffer J, Pang MG, Hoegerman SF, Osgood CJ, Stacey MW, Mayer J, et al. Aneuploidy frequencies in semen fractions from ten oligoasthenoteratozoospermic patients donating sperm for intracytoplasmic sperm injection. Fertil Steril. 1999;72(3):472-8.

Patassini C, Garolla A, Bottacin A, Menegazzo M, Speltra E, Foresta C, et al. Molecular karyotyping of human single sperm by array-comparative genomic hybridization. PLoS One. 2013;8(4):e60922.

Garolla A, Sartini B, Cosci I, Pizzol D, Ghezzi M, Bertoldo A, et al. Molecular karyotyping of single sperm with nuclear vacuoles identifies more chromosomal abnormalities in patients with testiculopathy than fertile controls: implications for ICSI. Hum Reprod. 2015;30(11):2493-500.

Ghoraeian P, Mozdarani H, Aleyasin A, Alizadeh-Nili H. Frequency of sex chromosomal disomy in spermatozoa of normal and oligozoospermic Iranian patients and its effects on fertilisation and implantation rates after ICSI. Andrologia. 2013;45(1):46-55.

Mihajlović AI, FitzHarris G. Segregating chromosomes in the mammalian oocyte. Curr Biol. 2018;28(16):R895-907.

Nakhuda G, Jing C, Butler R, Guimond C, Hitkari J, Taylor E, et al. Frequencies of chromosome-specific mosaicisms in trophoectoderm biopsies detected by next-generation sequencing. Fertil Steril. 2018;109(5):857-65.

Fishel S, Craig A, Lynch C, Dowell K, Ndukwe G, Jenner L, et al. Assessment of 19,803 paired chromosomes and clinical outcome from first 150 cycles using array CGH of the first polar body for embryo selection and transfer. J Fertil In Vitro. 2011;1(1):1-8.

Vicari E, de Palma A, Burrello N, Longo G, Grazioso C, Barone N, et al. Absolute polymorphic teratozoospermia in patients with oligo‐asthenozoospermia is associated with an elevated sperm aneuploidy rate. J Androl. 2003;24(4):598-603.

Ferlin A, Garolla A, Foresta C. Chromosome abnormalities in sperm of individuals with constitutional sex chromosomal abnormalities. Cytogenet Genome Res. 2005;111(3-4):310-6.

Moemen MN, Mostafa T, Gadalla AM, Abbas M, Ismail HF, Abd El‐Hamid MF, et al. Sperm disomy in idiopathic severely oligoasthenoteratozoospermic males. Andrologia. 2008;40(6):381-6.

Burrello N, Arcidiacono G, Vicari E, Asero P, Di Benedetto D, De Palma A, et al. Morphologically normal spermatozoa of patients with secretory oligo-astheno-teratozoospermia have an increased aneuploidy rate. Hum Reprod. 2004;19(10):2298-302.

Tran QT, Jatsenko T, Poolamets O, Tšuiko O, Lubenets D, Reimand T, et al. Chromosomal scan of single sperm cells by combining fluorescence-activated cell sorting and next-generation sequencing. J Assist Reprod Genet. 2019;36(1):91-7.

Sha Y, Sha Y, Ji Z, Ding L, Zhang Q, Ouyang H, et al. Comprehensive genome profiling of single sperm cells by multiple annealing and looping‐based amplification cycles and next‐generation sequencing from carriers of robertsonian translocation. Ann Hum Genet. 2017;81(2):91-7.

Magli MC, Gianaroli L, Ferraretti AP, Gordts S, Fredericks V, Crippa A. Paternal contribution to aneuploidy in preimplantation embryos. Reprod Biomed Online. 2009;18(4):536-42.

Coates A, Hesla JS, Hurliman A, Coate B, Holmes E, Matthews R, et al. Use of suboptimal sperm increases the risk of aneuploidy of the sex chromosomes in preimplantation blastocyst embryos. Fertil Steril. 2015;104(4):866-72.

Rodrigo L, Meseguer M, Mateu E, Mercader A, Peinado V, Bori L et al, Sperm chromosomal abnormalities and their contribution to human embryo aneuploidy. Biol Reprod. 2019;101(6):1091-101.