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Achieving an optimal pregnancy outcome through the combined utilization of micro-TESE and ICSI in cryptorchidism associated with a non-canonical splicing variant in RXFP2

  • Genetics
  • Published:
Journal of Assisted Reproduction and Genetics Aims and scope Submit manuscript

Abstract

Purpose

To identify the genetic cause of a cryptorchidism patient carrying a non-canonical splicing variant highlighted by SPCards platform in RXFP2 and to provide a comprehensive overview of RXFP2 variants with cryptorchidism correlation.

Methods

We identified a homozygous non-canonical splicing variant by whole-exome sequencing and Sanger sequencing in a case with cryptorchidism and non-obstructive azoospermia (NOA). As the pathogenicity of this non-canonical splicing variant remained unclear, we initially utilized the SPCards platform to predict its pathogenicity. Subsequently, we employed a minigene splicing assay to further evaluate the influence of the identified splicing variant. Microdissection testicular sperm extraction (micro-TESE) combined with intracytoplasmic sperm injection (ICSI) was performed. PubMed and Human Genome Variant Database (HGMD) were queried to search for RXFP2 variants.

Results

We identified a homozygous non-canonical splicing variant (NM_130806: c.1376-12A > G) in RXFP2, and confirmed this variant caused aberrant splicing of exons 15 and 16 of the RXFP2 gene: 11 bases were added in front of exon 16, leading to an abnormal transcript initiation and a frameshift. Fortunately, the patient successfully obtained his biological offspring through micro-TESE combined with ICSI. Four cryptorchidism-associated variants in RXFP2 from 90 patients with cryptorchidism were identified through a literature search in PubMed and HGMD, with different inheritance patterns.

Conclusion

This is the first cryptorchidism case carrying a novel causative non-canonical splicing RXFP2 variant. The combined approach of micro-TESE and ICSI contributed to an optimal pregnancy outcome. Our literature review demonstrated that RXFP2 variants caused cryptorchidism in a recessive inheritance pattern, rather than a dominant pattern.

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Data availability

The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request, without undue reservation.

References

  1. Gurney JK, McGlynn KA, Stanley J, Merriman T, Signal V, Shaw C, et al. Risk factors for cryptorchidism. Nat Rev Urol. 2017;14(9):534–48.

    Article  PubMed  PubMed Central  Google Scholar 

  2. Fantasia J, Aidlen J, Lathrop W, Ellsworth P. Undescended testes: a clinical and surgical review. Urologic Nursing. 2015;35(3):117–26.

    Article  PubMed  Google Scholar 

  3. Virtanen HE, Toppari J. Epidemiology and pathogenesis of cryptorchidism. Hum Reprod Update. 2008;14(1):49–58.

    Article  CAS  PubMed  Google Scholar 

  4. Dicke AK, Albrethsen J, Hoare BL, Wyrwoll MJ, Busch AS, Fietz D, et al. Bi-allelic variants in INSL3 and RXFP2 cause bilateral cryptorchidism and male infertility. Hum Reprod. 2023;38(7):1412–23.

    Article  CAS  PubMed  Google Scholar 

  5. Ferlin A, Simonato M, Bartoloni L, Rizzo G, Bettella A, Dottorini T, et al. The INSL3-LGR8/GREAT ligand-receptor pair in human cryptorchidism. J Clin Endocrinol Metab. 2003;88(9):4273–9.

    Article  CAS  PubMed  Google Scholar 

  6. Elamo HP, Virtanen HE, Toppari J. Genetics of cryptorchidism and testicular regression. Best Pract Res Clin Endocrinol Metab. 2022;36(1):101619.

    Article  CAS  PubMed  Google Scholar 

  7. Kong RC, Shilling PJ, Lobb DK, Gooley PR, Bathgate RA. Membrane receptors: structure and function of the relaxin family peptide receptors. Mol Cell Endocrinol. 2010;320(1–2):1–15.

    Article  CAS  PubMed  Google Scholar 

  8. Ivell R, Mamsen LS, Andersen CY, Anand-Ivell R. Expression and role of INSL3 in the fetal testis. Front Endocrinol (Lausanne). 2022;13:868313.

    Article  PubMed  Google Scholar 

  9. Gorlov IP, Kamat A, Bogatcheva NV, Jones E, Lamb DJ, Truong A, et al. mutations of the GREAT gene cause cryptorchidism. Hum Mol Genet. 2002;11(19):2309–18.

    Article  CAS  PubMed  Google Scholar 

  10. Bogatcheva NV, Ferlin A, Feng S, Truong A, Gianesello L, Foresta C, et al. T222P mutation of the insulin-like 3 hormone receptor LGR8 is associated with testicular maldescent and hinders receptor expression on the cell surface membrane. Am J Physiol Endocrinol Metab. 2007;292(1):E138–44.

    Article  CAS  PubMed  Google Scholar 

  11. Ferlin A, Zuccarello D, Zuccarello B, Chirico MR, Zanon GF, Foresta C. Genetic alterations associated with cryptorchidism. JAMA. 2008;300(19):2271–6.

    Article  CAS  PubMed  Google Scholar 

  12. El Houate B, Rouba H, Imken L, Sibai H, Chafik A, Boulouiz R, et al. No association between T222P/LGR8 mutation and cryptorchidism in the Moroccan population. Horm Res. 2008;70(4):236–9.

    PubMed  Google Scholar 

  13. Nuti F, Marinari E, Erdei E, El-Hamshari M, Echavarria MG, Ars E, et al. The leucine-rich repeat-containing G protein-coupled receptor 8 gene T222P mutation does not cause cryptorchidism. J Clin Endocrinol Metab. 2008;93(3):1072–6.

    Article  CAS  PubMed  Google Scholar 

  14. Ars E, Lo Giacco D, Bassas L, Nuti F, Rajmil O, Ruiz P, et al. Further insights into the role of T222P variant of RXFP2 in non-syndromic cryptorchidism in two Mediterranean populations. Int J Androl. 2011;34(4):333–8.

    Article  CAS  PubMed  Google Scholar 

  15. Tenenbaum-Rakover Y, Admoni O, Elias-Assad G, London S, Noufi-Barhoum M, Ludar H, et al. The evolving role of whole-exome sequencing in the management of disorders of sex development. Endocr Connect. 2021;10(6):620–9.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Ayers K, Kumar R, Robevska G, Bruell S, Bell K, Malik MA, et al. Familial bilateral cryptorchidism is caused by recessive variants in RXFP2. J Med Genet. 2019;56(11):727–33.

    Article  CAS  PubMed  Google Scholar 

  17. Osaka A, Iwahata T, Kobori Y, Shimomura Y, Yoshikawa N, Onota S, et al. Testicular volume in non-obstructive azoospermia with a history of bilateral cryptorchidism may predict successful sperm retrieval by testicular sperm extraction. Reprod Med Biol. 2020;19(4):372–7.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Mieusset R, Fouda PJ, Vaysse P, Guitard J, Moscovici J, Juskiewenski S. Increase in testicular temperature in case of cryptorchidism in boys. Fertil Steril. 1993;59(6):1319–21.

    Article  CAS  PubMed  Google Scholar 

  19. Minhas S, Bettocchi C, Boeri L, Capogrosso P, Carvalho J, Cilesiz NC, et al. European association of urology guidelines on male sexual and reproductive health: 2021 update on male infertility. Eur Urol. 2021;80(5):603–20.

    Article  PubMed  Google Scholar 

  20. Tang F, Gao Y, Li K, Tang D, Hao Y, Lv M, et al. Novel deleterious splicing variant in HFM1 causes gametogenesis defect and recurrent implantation failure: concerning the risk of chromosomal abnormalities in embryos. J Assist Reprod Genet. 2023;40(7):1689–702.

    Article  PubMed  Google Scholar 

  21. Tang D, Li K, Geng H, Xu C, Lv M, Gao Y, et al. Identification of deleterious variants in patients with male infertility due to idiopathic non-obstructive azoospermia. Reprod Biol Endocrinol. 2022;20(1):63.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Tang D, Lv M, Gao Y, Cheng H, Li K, Xu C, et al. Novel variants in helicase for meiosis 1 lead to male infertility due to non-obstructive azoospermia. Reprod Biol Endocrinol. 2021;19(1):129.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Ramasamy R, Reifsnyder JE, Husseini J, Eid PA, Bryson C, Schlegel PN. Localization of sperm during microdissection testicular sperm extraction in men with nonobstructive azoospermia. J Urol. 2013;189(2):643–6.

    Article  PubMed  Google Scholar 

  24. Carrier L, Schlossarek S, Willis MS, Eschenhagen T. The ubiquitin-proteasome system and nonsense-mediated mRNA decay in hypertrophic cardiomyopathy. Cardiovasc Res. 2010;85(2):330–8.

    Article  CAS  PubMed  Google Scholar 

  25. Kato K, Ohno S, Sonoda K, Fukuyama M, Makiyama T, Ozawa T, et al. LMNA missense mutation causes nonsense-mediated mRNA decay and severe dilated cardiomyopathy. Circ Genom Precis Med. 2020;13(5):435–43.

    Article  CAS  PubMed  Google Scholar 

  26. Wright CJ, Smith CWJ, Jiggins CD. Alternative splicing as a source of phenotypic diversity. Nat Rev Genet. 2022;23(11):697–710.

    Article  CAS  PubMed  Google Scholar 

  27. Sanders SJ, Schwartz GB, Farh KK. Clinical impact of splicing in neurodevelopmental disorders. Genome Med. 2020;12(1):36.

    Article  PubMed  PubMed Central  Google Scholar 

  28. Zhang S, Samocha KE, Rivas MA, Karczewski KJ, Daly E, Schmandt B, et al. Base-specific mutational intolerance near splice sites clarifies the role of nonessential splice nucleotides. Genome Res. 2018;28(7):968–74.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Lord J, Gallone G, Short PJ, McRae JF, Ironfield H, Wynn EH, et al. Pathogenicity and selective constraint on variation near splice sites. Genome Res. 2019;29(2):159–70.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Li K, Luo T, Zhu Y, Huang Y, Wang A, Zhang D, et al. Performance evaluation of differential splicing analysis methods and splicing analytics platform construction. Nucleic Acids Res. 2022;50(16):9115–26.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Lee PA, O’Leary LA, Songer NJ, Coughlin MT, Bellinger MF, LaPorte RE. Paternity after unilateral cryptorchidism: a controlled study. Pediatrics. 1996;98(4 Pt 1):676–9.

    Article  CAS  PubMed  Google Scholar 

  32. Jensen MS, Olsen LH, Thulstrup AM, Bonde JP, Olsen J, Henriksen TB. Age at cryptorchidism diagnosis and orchiopexy in Denmark: a population based study of 508,964 boys born from 1995 to 2009. J Urol. 2011;186(4 Suppl):1595–600.

    Article  PubMed  Google Scholar 

  33. Canavese F, Mussa A, Manenti M, Cortese MG, Ferrero L, Tuli G, et al. Sperm count of young men surgically treated for cryptorchidism in the first and second year of life: fertility is better in children treated at a younger age. Eur J Pediatr Surg. 2009;19(6):388–91.

    Article  CAS  PubMed  Google Scholar 

  34. C. Radmayr. EAU guidelines on paediatric urology. In: the EAU Guidelines 2023. European Association of Urology. 2023. https://uroweb.org/guidelines/paediatric-urology. Accessed 12 Aug 2023.

  35. Alrabeeah K, Wachter A, Phillips S, Cohen B, Al-Hathal N, Zini A. Sperm retrieval outcomes with microdissection testicular sperm extraction (micro-TESE) in men with cryptozoospermia. Andrology. 2015;3(3):462–6.

    Article  CAS  PubMed  Google Scholar 

  36. Bryson CF, Ramasamy R, Sheehan M, Palermo GD, Rosenwaks Z, Schlegel PN. Severe testicular atrophy does not affect the success of microdissection testicular sperm extraction. J Urol. 2014;191(1):175–8.

    Article  PubMed  Google Scholar 

  37. Chen XL, Wei YA, Ren XH, Zhang X, Li GY, Lu ZW, et al. Predictive factors for successful sperm retrieval by microdissection testicular sperm extraction in men with nonobstructive azoospermia and a history of cryptorchidism. Asian J Androl. 2022;24(5):503–8.

    Article  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

We would like to thank all the individuals who participated in and supported this research.

Funding

This study was supported by the National Natural Science Foundation of China (No. 82101681, 82071705, and 82101944) and the China Postdoctoral Science Foundation (2023M730019).

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Authors and Affiliations

  1. Reproductive Medicine Center, Department of Obstetrics and Gynecology, the First Affiliated Hospital of Anhui Medical University, No 218 Jixi Road, Hefei, 230022, Anhui, China

    Lewen Ruan, Meng Gu, Hao Geng, Zongliu Duan, Mingrong Lv & Dongdong Tang

  2. NHC Key Laboratory of Study On Abnormal Gametes and Reproductive Tract (Anhui Medical University), No 81 Meishan Road, Hefei, 230032, Anhui, China

    Lewen Ruan, Meng Gu, Hao Geng, Hui Yu, Zhongmei Shao, Kuokuo Li, Mingrong Lv & Dongdong Tang

  3. Key Laboratory of Population Health Across Life Cycle (Anhui Medical University), Ministry of Education of the People’s Republic of China, No 81 Meishan Road, Hefei, 230032, Anhui, China

    Lewen Ruan, Meng Gu, Hao Geng, Hui Yu, Zhongmei Shao, Kuokuo Li, Mingrong Lv & Dongdong Tang

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  1. Lewen Ruan
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Contributions

DT, KL, and ML designed the study. ZD and HY collected the samples and data. ZS and KL performed the data analysis. MG, HY, and ZS were responsible for in vitro experimentations. DT and HG were responsible for surgical operations. LR and HG wrote the paper. All authors have read and approved the final manuscript.

Corresponding authors

Correspondence to Kuokuo Li, Mingrong Lv or Dongdong Tang.

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The author declare no competing interests.

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Lewen Ruan, Meng Gu, and Hao Geng contributed equally to this work and shared first authorship.

Supplementary Information

Below is the link to the electronic supplementary material.

Supplementary file1 (DOCX 11 KB) Online Source 1 Primer information and expected size of PCR products.

Supplementary file2 (XLSX 49 KB) Online Source 2 Genetic information of candidate causal genes.

10815_2024_3070_MOESM3_ESM.pdf

Supplementary file3 (PDF 6 KB) Online Source 3 Results of homozygosity mapping. We have employed Automap (https://automap.iob.ch/) for homology mapping of the patient. The blue vertical lines represent the homozygous regions, while the black lines indicate the boundaries of a homozygous region: chr13-30764037-45465272. The red line indicates the location of the homozygous variant c.1376-12A>G identified in this study.

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Ruan, L., Gu, M., Geng, H. et al. Achieving an optimal pregnancy outcome through the combined utilization of micro-TESE and ICSI in cryptorchidism associated with a non-canonical splicing variant in RXFP2. J Assist Reprod Genet (2024). https://doi.org/10.1007/s10815-024-03070-4

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