Association of conventional chromosomal polymorphic variants in 580 couples with idiopathic recurrent pregnancy loss: A retrospective cohort study

Abstract

Objective: To investigate the potential link between chromosomal polymorphisms in couples who had a medical history of idiopathic recurrent pregnancy loss.

Methods: Cytogenetic investigation was conducted with mitogen (Phytohemagglutinin-M, Gibco) stimulated blood T lymphocytes by Giemsa trypsin Giemsa banding and Ag-NOR banding on 580 couples with a history of idiopathic recurrent pregnancy loss and 240 couples from the general population. Thirty good chromosomal spreads were captured, karyotyped, and analyzed. The karyotypes were designated using the International System for Human Cytogenomic Nomenclature 2024. Pearson Chi-square test was used to compare the frequency of chromosomal polymorphism variations in the idiopathic recurrent pregnancy loss group with the general population group.

Results: A conventional cytogenetic investigation revealed that 45.43% of couples experiencing idiopathic recurrent pregnancy loss presented with various types of chromosomal polymorphic variants, compared to 11.88% in the general population. The overall frequency of these chromosomal polymorphic variants was significantly higher in the idiopathic recurrent pregnancy loss group compared to the general population group (OR 9.97, 95% CI 6.9914.21; P<0.05). Additionally, the prevalence of polymorphic variants was higher among males (49.14%) than females (41.72%) (P=0.01).

Conclusions: Chromosomal polymorphic analysis may play a crucial role in the assessment and careful clinical management of cases with idiopathic recurrent pregnancy loss, especially when no other conclusive reasons are identified during the initial evaluation. Therefore, heteromorphism should not be overlooked while investigating the causes of idiopathic recurrent pregnancy loss.

Keywords

Polymorphic variants / Chromosomal anomalies / Repeated pregnancy loss / Cytogenetic investigation

Conflict of interest statement

The authors have no competing interests to declare.

Funding

The research was funded by the Technology Development Board (TDB) of India's Ministry of Science and Technology (TDB/ M-25/2018-19).

Authors’ contributions

Sunny Kumar Jignesh Kumar Patel participated in conception of the work, drafting the article, data analysis and interpretation; Amit Roy Chowdhury participated in critical revision of the article and final approval for publication; Shagufa Sheikh performed conventional cytogenetic technique and data curation; Purna Chandra Mahapatra contributed to investigation, data acquisition, and validation; Birendranath Banerjee participated in critical revision of the article and final approval for publication. All authors read and approved the final manuscript.

Publisher’ s Note

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References

[1] Fauzdar A, Dixit V, Mavi R, Khan T, Arya R, Sharma NK. High prevalence of D/G group acrocentric RPL chromosome polymorphisms in 1 400 recurrent pregnancy losses patients, an evaluation of genetic factor and reassessing CPMs in 21st century as normal variants? Int J Reprod Contracept Obstet Gynecol 2024; 13(3): 635.

[2] Genovese HG, McQueen DB. The prevalence of sporadic and recurrent pregnancy loss. Fertil Steril 2023 120(5): 934-936.

[3] Arias-Sosa LA, Acosta ID, Lucena-Quevedo E, Moreno-Ortiz H, Esteban-Pérez C, Forero-Castro M. Genetic and epigenetic variations associated with idiopathic recurrent pregnancy loss. J Assist Reprod Genet 2018; 35: 355-366.

[4] Turesheva A, Aimagambetova G, Ukybassova T, Marat A, Kanabekova P, Kaldygulova L, et al. Recurrent pregnancy loss etiology, risk factors, diagnosis, and management. Fresh look into a full box. J Clin Med 2023 12(12): 4074.

[5] Cao M, Zhang Q, Zhou W, Zhu Y, Li H, Yan J. Analysis of aneuploidy rate and pregnancy outcomes in unexplained recurrent pregnancy loss couples with chromosome polymorphism after PGT-A. Front Med 2022; 9: 803988.

[6] Alibakhshi R, Nejati P, Hamani S, Mir-Ahadi N, Jalilian N. Cytogenetic analysis of 570 couples with recurrent pregnancy loss: Reporting 11 years of experience. J Hum Reprod Sci 2020 13(3): 216-220.

[7] Kocaaga A, Kilic H, Gulec S. The pattern of chromosomal abnormalities in recurrent miscarriages: A single center retrospective study. Ann Saudi Med 2022 42(6): 385-390.

[8] Pohjoranta E, Mentula M, Gissler M, Suhonen S, Heikinheimo O. Provision of intrauterine contraception in association with first trimester induced abortion reduces the need of repeat abortion: First-year results of a randomized controlled trial. Hum Reprod 2015 30(11): 2539-2546.

[9] Tamaru H. Confining euchromatin/heterochromatin territory: Jumonji crosses the line. Genes Dev 2010 24(14): 1465-1478.

[10] Akbaş H, Isi H, Oral D, Türkyılmaz A, Kalkanlı-Taş S, Simşek S, et al. Chromosome heteromorphisms are more frequent in couples with recurrent abortions. Genet Mol Res 2012 11(4): 3847-3851.

[11] Mottola F, Santonastaso M, Ronga V, Finelli R, Rocco L. Polymorphic rearrangements of human chromosome 9 and male infertility: New evidence and impact on spermatogenesis. Biomolecules 2023 13(5): 729.

[12] Elhady GM, Kholeif S, Nazmy N. Chromosomal aberrations in 224 couples with recurrent pregnancy loss. J Hum Reprod Sci 2020 13(4): 340-348.

[13] Sheth FJ, Liehr T, Kumari P, Akinde R, Sheth HJ, Sheth JJ. Chromosomal abnormalities in couples with repeated fetal loss: An Indian retrospective study. Indian J Hum Genet 2013 19(4): 415.

[14] Alibakhshi R, Nejati P, Hamani S, Mir-Ahadi N, Jalilian N. Cytogenetic analysis of 570 couples with recurrent pregnancy loss: Reporting 11 years of experience. J Hum Reprod Sci 2020 13(3): 216-220.

[15] Bender Atik R, Christiansen OB, Elson J, Kolte AM, Lewis S, Middeldorp S, et al. ESHRE guideline: Recurrent pregnancy loss: An update in 2022. Hum Reprod Open 2023; 2023(1): hoad002. doi: 10.1093/hropen/hoad002.

[16] Toth B, Würfel W, Bohlmann M, Zschocke J, Rudnik-Schöneborn S, Nawroth F, et al. Recurrent miscarriage: Diagnostic and therapeutic procedures. Guideline of the DGGG, OEGGG and SGGG (S2k-Level, AWMF Registry Number 015/050). Geburtshilfe Frauenheilkd 2018; 78(4): 364-381. doi: 10.1055/a-0586-4568.

[17] Priya PK, Mishra VV, Roy P, Patel H. A study on balanced chromosomal translocations in couples with recurrent pregnancy loss. Hum Reprod Sci 2018 11(4): 337-342.

[18] Rawal L, Kumar S, Mishra SR, Lal V, Bhattacharya SK. Clinical manifestations of chromosomal anomalies and polymorphic variations in patients suffering from reproductive failure. Hum Reprod Sci 2020 13(3): 209-215.

[19] Iyer P, Wani L, Joshi S, Lakshmi J, Dalvi R, Chavan D, et al. Cytogenetic investigations in couples with repeated miscarriages and malformed children: Report of a novel insertion. Reprod Biomed 2007 14(3): 314-321.

[20] Minocherhomji S, Athalye AS, Madon PF, Kulkarni D, Uttamchandani SA, Parikh FR. A case-control study identifying chromosomal polymorphic variations as forms of epigenetic alterations associated with the infertility phenotype. Fertil Steril 2009 92(1): 88-95.

[21] Madon PF, Athalye AS, Parikh FR. Polymorphic variants on chromosomes probably play a significant role in infertility. Reprod Biomed 2005 11(6): 726-732.

[22] Akbaş E, Yazıcı FG, Durukan H, Topal H, Erdoğan NE. Cytogenetic and clinical evaluation of two cases with 45, X/46, X, i (Xq) and 46, X, i (Xq) karyotype. J Clin Exp Invest 2014; 5(3). doi: 10.5799/ahinjs.01.2014.03.0436.

[23] Wang Y, Li G, Zuo MZ, Fang JH, Li HR, Quan DD, et al. Y chromosome polymorphisms may contribute to an increased risk of male-induced unexplained recurrent miscarriage. Biosci Rep 2017; 37(2): BSR20160528. doi: 10.1042/BSR20160528.

[24] Imam SN, Shamsi MB, Kumar K, Deka D, Dada R. Idiopathic recurrent pregnancy loss: Role of paternal factors; a pilot study. J Reprod Infertil 2011 12(4): 267.

[25] Caglayan AO, Ozyazgan I, Demiryilmaz F, Ozgun MT. Are heterochromatin polymorphisms associated with recurrent miscarriage? J Obstet Gynaecol Res 2010; 36(4): 774-776.

[26] Christofolini DM, Mafra FA, Neto RP, Saab de Almeida Barros RA, Amaro dos Santos A, Peluso C, et al. Correlation between chromosomal variants and male infertility in a population of Brazilian infertile men. Reprod Syst Sex Dis 2012 1(1): 2-6.

[27] Humphray SJ, Oliver K, Hunt AR, Plumb RW, Loveland JE, Howe KL, et al. DNA sequence and analysis of human chromosome 9. Nature 2004 429(6990): 369-374.

[28] Šípek Jr A, Mihalová R, Panczak A, Hrčková L, Janashia M, Kaspříková N, et al. Heterochromatin variants in human karyotypes: A possible association with reproductive failure. Reprod Biomed Online 2014 29(2): 245-250.

[29] Feng X, Liu J, Wang Y, Fu J, Qin Q, Cao Y, et al. Acrocentric chromosome polymorphic variants on Chinese female have possible association with unexplained recurrent pregnancy loss. Reprod Sci 2021; 28: 575-584.

[30] Sheth FJ, Shah UJ, Desai MJ, Sheth JJ. Clinical profile of inversion Y in people of Gujarat, West India. Int J Hum Genet 2011 11(4): 245-248.

[31] Atlı Eİ, Mail Ç, Gurkan H, Yalçıntepe S, Demir S, Atlı E. Y chromosome polymorphism in Turkish patients with reproductive problems: A genetic centre experience. Eur J Res 2023 9(4): 725-729.

[32] Mullen TJ, Davis-Roca AC, Wignall SM. Spindle assembly and chromosome dynamics during oocyte meiosis. Curr Opin Cell Biol 2019; 60: 53-59.

[33] Chakraborty A, Kar S, Mohapatra PC, Banerjee B. A case-control study identifying the frequency and spectrum of chromosomal anomalies and variants in a cohort of 1000 couples with a known history of recurrent pregnancy loss in the Eastern region of India. J Hum Reprod Sci 2021 14(4): 422-430.

[34] Chopade S, Chopade DK, Harde H. Impact of chromosomal heteromorphisms on recurrent miscarriages. Hum Genet Embryol 2012; 2: 101.

[35] Mukhopadhyay S, Dasgupta S, Ghosh K, Mukherjee T. Investigating the relation between chromosomal polymorphism and recurrent pregnancy loss: A cohort study. Indian J Obstet Gynecol Res 2023 9(3): 391-396.

[36] Kalotra V, Lall M, Saviour P, Verma IC, Kaur A. Prevalence of cytogenetic anomalies in couples with recurrent miscarriages: A case-control study. Indian J Obstet Gynecol Res 2017 10(4): 302-309.

[37] Morales R, Lledó B, Ortiz JA, Ten J, Llácer J, Bernabeu R. Chromosomal polymorphic variants increase aneuploidies in male gametes and embryos. Syst Biol Reprod Med 2016; 62(5): 317-324. doi: 10.1080/19396368.2016.1212949.

[38] Mau UA, Bäckert IT, Kaiser P, Kiesel L. Chromosomal findings in 150 couples referred for genetic counselling prior to intracytoplasmic sperm injection. Hum Reprod 1997 12(5): 930-937.

[39] Rizzi N, Denegri M, Chiodi I, Corioni M, Valgardsdottir R, Cobianchi F, et al. Transcriptional activation of a constitutive heterochromatic domain of the human genome in response to heat shock. Mol Biol Cell 2004 15(2): 543-551.