Skip to main content
SpringerLink
Log in

Normalized Mitochondrial DNA Copy Number Can Optimize Pregnancy Outcome Prediction in IVF

  • Embryology: Original Article
  • Published:
Reproductive Sciences Aims and scope Submit manuscript

Abstract

The aim of this study is to explore the relationship between mitochondrial DNA (mtDNA) copy number and embryo implantation potential in in vitro fertilization (IVF). A retrospective study of 319 blastocysts from patients undergoing preimplantation genetic testing (PGT) at Reproductive Medicine Center in Tongji Hospital from January 2016 to February 2018 was conducted. We used multiple annealing- and looping-based amplification cycles (MALBAC) technology to amplify the genetic materials from the trophectoderm cells of blastocysts, and next-generation sequencing (NGS) technology to test mitochondrial DNA copy number. Box-Cox transformation was introduced to eliminate the skewness distribution of mtDNA copy number, and the transformed data were defined as adjusted mtDNA. Subsequently, associations between adjusted mtDNA and the clinical characteristics of patients were assessed by univariate analysis and multiple linear regression. In addition, Gaussian Naive Bayes classifier was also used to predict pregnancy outcomes. We observed that only antral follicle count (AFC) was significantly associated with adjusted mtDNA without the influence of multicollinearity. What’s more, the distribution of the adjusted mtDNA of blastocysts resulting in live birth was more concentrated than that of others. The area under the curve (AUC) of the prediction model that combined adjusted mtDNA with other clinical characteristics of patients was up to 0.81, higher than that excluded adjusted mtDNA. Among patient clinical characteristics, AFC was significantly associated with adjusted mtDNA. Mitochondrial DNA copy number may help to optimize the pregnancy outcome prediction in IVF.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  1. Ishihara O, Adamson GD, Dyer S, de Mouzon J, Nygren KG, Sullivan EA, et al. International committee for monitoring assisted reproductive technologies: world report on assisted reproductive technologies, 2007. Fertil Steril. 2015;103:402–13.e11.

    Article  Google Scholar 

  2. Alpha Scientists in Reproductive Medicine and ESHRE Special Interest Group of Embryology. The Istanbul consensus workshop on embryo assessment: proceedings of an expert meeting. Hum Reprod. 2011;26:1270–83.

    Article  Google Scholar 

  3. Forman EJ, Hong KH, Ferry KM, Tao X, Taylor D, Levy B, et al. In vitro fertilization with single euploid blastocyst transfer: a randomized controlled trial. Fertil Steril. 2013;100:100–7.e1.

    PubMed  Google Scholar 

  4. Rubio C, Bellver J, Rodrigo L, Castillón G, Guillén A, Vidal C, et al. In vitro fertilization with preimplantation genetic diagnosis for aneuploidies in advanced maternal age: a randomized, controlled study. Fertil Steril. 2017;107:1122–9.

    Article  Google Scholar 

  5. Scott RT, Upham KM, Forman EJ, Hong KH, Scott KL, Taylor D, et al. Blastocyst biopsy with comprehensive chromosome screening and fresh embryo transfer significantly increases in vitro fertilization implantation and delivery rates: a randomized controlled trial. Fertil Steril. 2013;100:697–703.

    Article  Google Scholar 

  6. Simon AL, Kiehl M, Fischer E, Proctor JG, Bush MR, Givens C, et al. Pregnancy outcomes from more than 1,800 in vitro fertilization cycles with the use of 24-chromosome single-nucleotide polymorphism-based preimplantation genetic testing for aneuploidy. Fertil Steril. 2018;110:113–21.

    Article  Google Scholar 

  7. Fragouli E, McCaffrey C, Ravichandran K, Spath K, Grifo JA, Munné S, et al. Clinical implications of mitochondrial DNA quantification on pregnancy outcomes: a blinded prospective non-selection study. Hum Reprod. 2017;32:2340–7.

    Article  CAS  Google Scholar 

  8. Fragouli E, Spath K, Alfarawati S, Kaper F, Craig A, Michel CE, et al. Altered levels of mitochondrial DNA are associated with female age, aneuploidy, and provide an independent measure of embryonic implantation potential. PLoS Genet. 2015;11:e1005241.

    Article  Google Scholar 

  9. May-Panloup P, Chrétien MF, Savagner F, Vasseur C, Jean M, Malthièry Y, et al. Increased sperm mitochondrial DNA content in male infertility. Hum Reprod. 2003;18:550–6.

    Article  CAS  Google Scholar 

  10. van Blerkom J, Davis PW, Lee J. ATP content of human oocytes and developmental potential and outcome after in-vitro fertilization and embryo transfer. Hum Reprod. 1995;10:415–24.

    Article  CAS  Google Scholar 

  11. van Blerkom J. Mitochondria in human oogenesis and preimplantation embryogenesis: engines of metabolism, ionic regulation and developmental competence. Reproduction. 2004;128:269–80.

    Article  Google Scholar 

  12. de Los Santos MJ, Diez Juan A, Mifsud A, Mercader A, Meseguer M, Rubio C, et al. Variables associated with mitochondrial copy number in human blastocysts: what can we learn from trophectoderm biopsies? Fertil Steril. 2018;109:110–7.

    Article  Google Scholar 

  13. Ho JR, Arrach N, Rhodes-Long K, Salem W, McGinnis LK, Chung K, et al. Blastulation timing is associated with differential mitochondrial content in euploid embryos. J Assist Reprod Genet. 2018;35:711–20.

    Article  Google Scholar 

  14. Tan Y, Yin X, Zhang S, Jiang H, Tan K, Li J, et al. Clinical outcome of preimplantation genetic diagnosis and screening using next generation sequencing. GigaScience. 2014;3:30.

    Article  Google Scholar 

  15. Victor AR, Brake AJ, Tyndall JC, Griffin DK, Zouves CG, Barnes FL, et al. Accurate quantitation of mitochondrial DNA reveals uniform levels in human blastocysts irrespective of ploidy, age, or implantation potential. Fertil Steril. 2017;107:34–42.e3.

    Article  Google Scholar 

  16. Diez-Juan A, Rubio C, Marin C, Martinez S, Al-Asmar N, Riboldi M, et al. Mitochondrial DNA content as a viability score in human euploid embryos: less is better. Fertil Steril. 2015;104:534–41.e1.

    Article  Google Scholar 

  17. Treff NR, Zhan Y, Tao X, Olcha M, Han M, Rajchel J, et al. Levels of trophectoderm mitochondrial DNA do not predict the reproductive potential of sibling embryos. Hum Reprod. 2017;32:954–62.

    CAS  PubMed  PubMed Central  Google Scholar 

  18. Humaidan P, Kristensen SG, Coetzee K. Mitochondrial DNA, a new biomarker of embryonic implantation potential: fact or fiction? Fertil Steril. 2018;109:61–2.

    Article  Google Scholar 

  19. Wells D. Mitochondrial DNA quantity as a biomarker for blastocyst implantation potential. Fertil Steril. 2017;108:742–7.

    Article  CAS  Google Scholar 

  20. Zegers-Hochschild F, Adamson GD, Dyer S, Racowsky C, de Mouzon J, Sokol R, et al. The International Glossary on Infertility and Fertility Care, 2017. Hum Reprod. 2017;32:1786–801.

    Article  Google Scholar 

  21. Zong C, Lu S, Chapman AR, Xie XS. Genome-wide detection of single-nucleotide and copy-number variations of a single human cell. Science. 2012;338:1622–6.

    Article  CAS  Google Scholar 

  22. Wells D, Ravichandran K, McCaffrey C, Grifo J, Morales A, Perloe M, et al. Reply: mitochondrial DNA quantification-the devil in the detail. Hum Reprod. 2017;32:2150–1.

    Article  CAS  Google Scholar 

  23. Lee Y, Chen C, Lin S, Lin Y, Tzeng C. Adjusted mitochondrial DNA quantification in human embryos may not be applicable as a biomarker of implantation potential. J Assist Reprod Genet. 2019;36(9):1855–65.

    Article  Google Scholar 

  24. Huang B, Qian K, Li Z, Yue J, Yang W, Zhu G, et al. Neonatal outcomes after early rescue intracytoplasmic sperm injection: an analysis of a 5-year period. Fertil Steril. 2015;103:1432–7.e1.

    PubMed  Google Scholar 

  25. Jin L, Wang M, Yue J, Zhu GJ, Zhang B. Association between TSH level and pregnancy outcomes in euthyroid women undergoing IVF/ICSI: a retrospective study and meta-analysis. Curr Med Sci. 2019;39:631–7.

    Article  CAS  Google Scholar 

  26. Hou Y, Fan W, Yan L, Li R, Lian Y, Huang J, et al. Genome analyses of single human oocytes. Cell. 2013;155:1492–506.

    Article  CAS  Google Scholar 

  27. Shang W, Zhang Y, Shu M, Wang W, Ren L, Chen F, et al. Comprehensive chromosomal and mitochondrial copy number profiling in human IVF embryos. Reprod BioMed Online. 2018;36:67–74.

    Article  CAS  Google Scholar 

  28. Yamaguchi Y, Maruo K, Partlett C, Riley RD. A random effects meta-analysis model with Box-Cox transformation. BMC Med Res Methodol. 2017;17:109.

    Article  Google Scholar 

  29. Joshi D, Mishra A, Anand S. A naïve Gaussian Bayes classifier for detection of mental activity in gait signature. Comput Methods Biomech Biomed Engin. 2012;15:411–6.

    Article  Google Scholar 

  30. Ravichandran K, McCaffrey C, Grifo J, Morales A, Perloe M, Munne S, et al. Mitochondrial DNA quantification as a tool for embryo viability assessment: retrospective analysis of data from single euploid blastocyst transfers. Hum Reprod. 2017;32:1282–92.

    Article  CAS  Google Scholar 

  31. Lu S, Zong C, Fan W, Yang M, Li J, Chapman AR, et al. Probing meiotic recombination and aneuploidy of single sperm cells by whole-genome sequencing. Science. 2012;338:1627–30.

    Article  CAS  Google Scholar 

  32. van Blerkom J. Mitochondrial function in the human oocyte and embryo and their role in developmental competence. Mitochondrion. 2011;11:797–813.

    Article  Google Scholar 

  33. Motta PM, Nottola SA, Makabe S, Heyn R. Mitochondrial morphology in human fetal and adult female germ cells. Hum Reprod. 2000;15:129–47.

    Article  Google Scholar 

  34. Sathananthan AH, Trounson AO. Mitochondrial morphology during preimplantational human embryogenesis. Hum Reprod. 2000;15:148–59.

    Article  Google Scholar 

  35. Lin DP, Huang CC, Wu HM, Cheng TC, Chen CI, Lee MS. Comparison of mitochondrial DNA contents in human embryos with good or poor morphology at the 8-cell stage. Fertil Steril. 2004;81:73–9.

    Article  CAS  Google Scholar 

  36. Houghton FD. Energy metabolism of the inner cell mass and trophectoderm of the mouse blastocyst. Differentiation. 2006;74:11–8.

    Article  CAS  Google Scholar 

Download references

Acknowledgments

We would like to express heartfelt gratitude to Yue Sun and his colleagues from Yikon Genomics for helping us process the data of the mitochondrial DNA during the MALBAC-NGS process. We also want to thank Yuqi Cui from Huazhong University of Science and Technology in China for helping in developing our machine learning algorithms.

Funding

This work was supported by the National Key Research and Development Project (2018YFC1002103) and the Chinese Medical Association (16020520668) as well as the Natural Science Foundation of Hubei Province (2017CFB752).

Author information

Authors and Affiliations

  1. Reproductive Medicine Center, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China

    Lixia Zhu, Jingjing Li, Meng Wang, Zishui Fang, Fangqin Zheng, Zhou Li & Lei Jin

Authors
  1. Lixia Zhu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jingjing Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Meng Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Zishui Fang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Fangqin Zheng
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Zhou Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Lei Jin
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

LJ and LZ conceived the study; JL and MW wrote the paper; ZF and ZL collected data; JL, MW, and FZ analyzed data.

Corresponding author

Correspondence to Lei Jin.

Ethics declarations

Competing Interests

The authors declare that they have no conflict of interest.

Ethics Approval and Consent to Participate

The study was approved by the Ethics of Committee of Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology. All the participants have signed the consent forms.

Additional information

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary Information

ESM 1

(DOCX 15 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Zhu, L., Li, J., Wang, M. et al. Normalized Mitochondrial DNA Copy Number Can Optimize Pregnancy Outcome Prediction in IVF. Reprod. Sci. 28, 1439–1446 (2021). https://doi.org/10.1007/s43032-020-00422-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s43032-020-00422-0

Keywords

Search

Navigation