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Journal of Assisted Reproduction and Genetics

, Volume 36, Issue 9, pp 1855–1865 | Cite as

Adjusted mitochondrial DNA quantification in human embryos may not be applicable as a biomarker of implantation potential

  • Yi-Xuan Lee
  • Chi-Huang Chen
  • Shyr-Yeu Lin
  • Yi-Hui Lin
  • Chii-Ruey TzengEmail author
Embryo Biology
  • 97 Downloads

Abstract

Objective

To evaluate the feasibility of adjusted mitochondrial DNA quantification in human embryos as a biomarker for implantation potential.

Design

Double-blind, observational, prospective analysis of an Asian population in a single university-affiliated in vitro fertilization center. A total of 1617 embryos derived from 380 infertile couples were collected. The DNA from blastomere biopsy (n = 99) or trophectoderm biopsy (n = 1518) were analyzed with next-generation sequencing.

Results

The adjusted mtDNA quantification followed a non-normal distribution in both types of the embryos. When stratified by ploidy status, the adjusted mtDNA quantification was significantly higher in aneuploid trophectoderm than in euploid cells, but not in blastomeres. The adjusted mtDNA quantification of embryos showed significant but very weak positive correlation in total trophectoderm cells with maternal age (Spearman’s correlation, r = 0.095, p = 0.0028) but neither in blastomeres nor stratified by ploidy status. The median adjusted mtDNA quantification was also significantly higher in aneuploid blastocysts than in euploid ones while corrected with embryo morphology. Viable embryos did not contain significantly different quantities of adjusted mtDNA compared with nonviable embryos (implanted n = 103, non-implanted n = 164; median 0.00097 vs. 0.00088, p = 0.21) in 267 transferred blastocysts.

Conclusion

Quantification of adjusted mitochondria DNA in human embryos was significantly lower in euploid blastocysts than in aneuploid blastocysts. However, no statistically significant differences regarding implantation outcome were evident. To our best knowledge, this study provides the largest scale and the first correlation data between mitochondria copy number and human embryo implantation potential in Asians.

Keywords

Mitochondrial DNA copy number Correction factors PGT-A IVF Biomarker 

Notes

Acknowledgments

The authors thank all the embryologists of the reproductive center of Taipei Medical University Hospital for embryo biopsy and sample preparation. We also thank the bioinformatics support provided by the Genomics Center for Clinical and Biotechnological Applications of the National Yang-Ming University VGH Genome Research Center. Genomics Center for Clinical and Biotechnological Applications is supported by the National Core Facility for Biopharmaceuticals, Ministry of Science and Technology. We also thank the Taipei Medical University Hospital for financial support. Finally, we thank the Genetics Generation Advancement Corporation for supporting this study in various forms, including sample preparation and interpretation, technical support, and extremely helpful discussions.

Funding

This study was partially supported by research grants NSC96-2314-B-038-019 and MOST-104-2314-B-038-063-MY2 from the Ministry of Science and Technology, Taiwan, Academia Sinica (BM10501010036, BM10601010024), National Healthy Research Institute (MG-1050SP-07, MG-106-SP-07), and Taipei Medical University Hospital (108TMUH-NE-01). The funders played no role in the conduct of the study or the writing of the manuscript.

Compliance with ethical standards

This study was reviewed and approved by the Taipei Medical University joint institutional review board (TMU-JIRB, approval number: N201707027). Patient informed consent for analysis of surplus DNA product was obtained with the approval of the ethical committee.

Conflict of interest

Dr. Yi-Hui Lin serves as a medical consultant for Genetics Generation Advancement Corporation. The other authors declare that they have no conflict of interest.

References

  1. 1.
    Chang J, Boulet SL, Jeng G, Flowers L, Kissin DM. Outcomes of in vitro fertilization with preimplantation genetic diagnosis: an analysis of the United States Assisted Reproductive Technology Surveillance Data, 2011–2012. Fertil Steril. 2016;105(2):394–400.CrossRefGoogle Scholar
  2. 2.
    Fragouli E, Alfarawati S, Spath K, Jaroudi S, Sarasa J, Enciso M, et al. The origin and impact of embryonic aneuploidy. Hum Genet. 2013;132(9):1001–13.CrossRefGoogle Scholar
  3. 3.
    Rosenwaks Z. Introduction: biomarkers of embryo viability: the search for the “holy grail” of embryo selection. Fertil Steril. 2017;108(5):719–21.CrossRefGoogle Scholar
  4. 4.
    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. Fertil Steril. 2017;108(3):393–406.CrossRefGoogle Scholar
  5. 5.
    Chan DC. Mitochondria: dynamic organelles in disease, aging, and development. Cell. 2006;125(7):1241–52.CrossRefGoogle Scholar
  6. 6.
    Wai T, Ao A, Zhang X, Cyr D, Dufort D, Shoubridge EA. The role of mitochondrial DNA copy number in mammalian fertility. Biol Reprod. 2010;83(1):52–62.CrossRefGoogle Scholar
  7. 7.
    May-Panloup P, Boucret L, Chao de la Barca JM, Desquiret-Dumas V, Ferré-L'Hotellier V, Morinière C, et al. Ovarian ageing: the role of mitochondria in oocytes and follicles. Hum Reprod Update. 2016;22(6):725–43.CrossRefGoogle Scholar
  8. 8.
    St John JC, et al. Mitochondrial DNA transmission, replication and inheritance: a journey from the gamete through the embryo and into offspring and embryonic stem cells. Hum Reprod Update. 2010;16(5):488–509.CrossRefGoogle Scholar
  9. 9.
    Hsieh R-H, Au HK, Yeh TS, Chang SJ, Cheng YF, Tzeng CR. Decreased expression of mitochondrial genes in human unfertilized oocytes and arrested embryos. Fertil Steril. 2004;81:912–8.CrossRefGoogle Scholar
  10. 10.
    Hsieh R-H, Tsai NM, Au HK, Chang SJ, Wei YH, Tzeng CR. Multiple rearrangements of mitochondrial DNA in unfertilized human oocytes. Fertil Steril. 2002;77(5):1012–7.CrossRefGoogle Scholar
  11. 11.
    Van Blerkom J. Mitochondria in human oogenesis and preimplantation embryogenesis: engines of metabolism, ionic regulation and developmental competence. Reproduction. 2004;128(3):269–80.CrossRefGoogle Scholar
  12. 12.
    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(6):e1005241.CrossRefGoogle Scholar
  13. 13.
    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(3):534–41 e1.CrossRefGoogle Scholar
  14. 14.
    Ravichandran K, et al. Mitochondrial DNA quantification as a tool for embryo viability assessment: retrospective analysis of data from single euploid blastocyst transfers. Hum Reprod. 2017:1–11.Google Scholar
  15. 15.
    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(11):2340–7.CrossRefGoogle Scholar
  16. 16.
    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(1):34–42 e3.CrossRefGoogle Scholar
  17. 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. 2017:1–9.Google Scholar
  18. 18.
    Fiorentino F, Bono S, Biricik A, Nuccitelli A, Cotroneo E, Cottone G, et al. Application of next-generation sequencing technology for comprehensive aneuploidy screening of blastocysts in clinical preimplantation genetic screening cycles. Hum Reprod. 2014;29(12):2802–13.CrossRefGoogle Scholar
  19. 19.
    Gardner DK, Lane M, Stevens J, Schlenker T, Schoolcraft WB. Blastocyst score affects implantation and pregnancy outcome: towards a single blastocyst transfer. Fertil Steril. 2000;73(6):1155–8.CrossRefGoogle Scholar
  20. 20.
    Nasiri N, Eftekhari-Yazdi P. An overview of the available methods for morphological scoring of pre-implantation embryos in in vitro fertilization. Cell J. 2015;16(4):392–405.Google Scholar
  21. 21.
    Scott RT, 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(3):697–703.CrossRefGoogle Scholar
  22. 22.
    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(5):1122–9.CrossRefGoogle Scholar
  23. 23.
    Practice Committees of the American Society for Reproductive M, et al. The use of preimplantation genetic testing for aneuploidy (PGT-A): a committee opinion. Fertil Steril. 2018;109(3):429–36.CrossRefGoogle Scholar
  24. 24.
    Capalbo A, Rienzi L, Cimadomo D, Maggiulli R, Elliott T, Wright G, et al. Correlation between standard blastocyst morphology, euploidy and implantation: an observational study in two centers involving 956 screened blastocysts. Hum Reprod. 2014;29(6):1173–81.CrossRefGoogle Scholar
  25. 25.
    Minasi MG, Colasante A, Riccio T, Ruberti A, Casciani V, Scarselli F, et al. Correlation between aneuploidy, standard morphology evaluation and morphokinetic development in 1730 biopsied blastocysts: a consecutive case series study. Hum Reprod. 2016;31(10):2245–54.CrossRefGoogle Scholar
  26. 26.
    de Los Santos MJ, et al. Variables associated with mitochondrial copy number in human blastocysts: what can we learn from trophectoderm biopsies? Fertil Steril. 2018;109(1):110–7.CrossRefGoogle Scholar
  27. 27.
    Wells D. Mitochondrial DNA quantity as a biomarker for blastocyst implantation potential. Fertil Steril. 2017;108(5):742–7.CrossRefGoogle Scholar
  28. 28.
    Humaidan P, Kristensen SG, Coetzee K. Mitochondrial DNA, a new biomarker of embryonic implantation potential: fact or fiction? Fertil Steril. 2018;109(1):61–2.CrossRefGoogle Scholar
  29. 29.
    Viotti M, Victor AR, Zouves CG, Barnes FL. Is mitochondrial DNA quantitation in blastocyst trophectoderm cells predictive of developmental competence and outcome in clinical IVF? J Assist Reprod Genet. 2017;34(12):1581–5.CrossRefGoogle Scholar
  30. 30.
    Scott RT Jr. Enhanced techniques to “power” embryonic mitochondria research. Fertil Steril. 2017;107(1):59–60.CrossRefGoogle Scholar
  31. 31.
    Barritt JA, K.M., Cohen J, Steuerwald N, Brenner CA, Quantification of human ooplasmic mitochondria. Reprod BioMed Online, 2002. 4: p. 243–247.Google Scholar
  32. 32.
    Lin D. Comparison of mitochondrial DNA contents in human embryos with good or poor morphology at the 8-cell stage. Fertil Steril. 2004;81(1):73–9.CrossRefGoogle Scholar
  33. 33.
    Eichenlaub-Ritter U, Wieczorek M, Lüke S, Seidel T. Age related changes in mitochondrial function and new approaches to study redox regulation in mammalian oocytes in response to age or maturation conditions. Mitochondrion. 2011;11(5):783–96.CrossRefGoogle Scholar
  34. 34.
    Tsai T, John JCS. The role of mitochondrial DNA copy number, variants, and haplotypes in farm animal developmental outcome. Domest Anim Endocrinol. 2016;56:S133–46.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Yi-Xuan Lee
    • 1
    • 2
  • Chi-Huang Chen
    • 2
    • 3
  • Shyr-Yeu Lin
    • 2
    • 3
  • Yi-Hui Lin
    • 4
    • 5
  • Chii-Ruey Tzeng
    • 2
    • 3
    • 6
    Email author
  1. 1.Graduate Institute of Clinical MedicineTaipei Medical UniversityTaipeiTaiwan
  2. 2.Division of Infertility, Department of Obstetrics and GynecologyTaipei Medical University HospitalTaipei CityTaiwan
  3. 3.Department of Obstetrics and Gynecology, School of Medicine, College of MedicineTaipei Medical UniversityTaipeiTaiwan
  4. 4.Department of Obstetrics and Gynecology, Wan Fang HospitalTaipei Medical UniversityTaipeiTaiwan
  5. 5.Genetics Generation Advancement Corporation (GGA Corp.)TaipeiTaiwan
  6. 6.Center for Reproductive MedicineTaipei Medical University HospitalTaipei CityTaiwan

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