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Pronuclear transfer in human embryos to prevent transmission of mitochondrial DNA disease

Nature volume 465, pages 82–85 (2010)Cite this article

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Abstract

Mutations in mitochondrial DNA (mtDNA) are a common cause of genetic disease. Pathogenic mutations in mtDNA are detected in approximately 1 in 250 live births1,2,3 and at least 1 in 10,000 adults in the UK are affected by mtDNA disease4. Treatment options for patients with mtDNA disease are extremely limited and are predominantly supportive in nature. Mitochondrial DNA is transmitted maternally and it has been proposed that nuclear transfer techniques may be an approach for the prevention of transmission of human mtDNA disease5,6. Here we show that transfer of pronuclei between abnormally fertilized human zygotes results in minimal carry-over of donor zygote mtDNA and is compatible with onward development to the blastocyst stage in vitro. By optimizing the procedure we found the average level of carry-over after transfer of two pronuclei is less than 2.0%, with many of the embryos containing no detectable donor mtDNA. We believe that pronuclear transfer between zygotes, as well as the recently described metaphase II spindle transfer, has the potential to prevent the transmission of mtDNA disease in humans.

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Figure 1: Pronuclear transfer using abnormally fertilized human zygotes.
Figure 2: Mitochondrial DNA analysis of pronuclear-transfer embryos.
Figure 3: Mitochondrial DNA analysis of individual blastomeres disaggregated from pronuclear-transfer embryos.

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References

  1. Elliott, H. R., Samuels, D. C., Eden, J. A., Relton, C. L. & Chinnery, P. F. Pathogenic mitochondrial DNA mutations are common in the general population. Am. J. Hum. Genet. 83, 254–260 (2008)

    Article  CAS  Google Scholar 

  2. Vandebona, H. et al. Prevalence of mitochondrial 1555A→G mutation in adults of European descent. N. Engl. J. Med. 360, 642–644 (2009)

    Article  CAS  Google Scholar 

  3. Bitner-Glindzicz, M. et al. Prevalence of mitochondrial 1555A→G mutation in European children. N. Engl. J. Med. 360, 640–642 (2009)

    Article  Google Scholar 

  4. Schaefer, A. M. et al. Prevalence of mitochondrial DNA disease in adults. Ann. Neurol. 63, 35–39 (2008)

    Article  CAS  Google Scholar 

  5. Sato, A. et al. Gene therapy for progeny of mito-mice carrying pathogenic mtDNA by nuclear transplantation. Proc. Natl Acad. Sci. USA 102, 16765–16770 (2005)

    Article  ADS  CAS  Google Scholar 

  6. Brown, D. T. et al. Transmission of mitochondrial DNA disorders: possibilities for the future. Lancet 368, 87–89 (2006)

    Article  CAS  Google Scholar 

  7. Wallace, D. C. Why do we still have a maternally inherited mitochondrial DNA? Insights from evolutionary medicine. Annu. Rev. Biochem. 76, 781–821 (2007)

    Article  CAS  Google Scholar 

  8. DiMauro, S. & Schon, E. A. Mitochondrial disorders in the nervous system. Annu. Rev. Neurosci. 31, 91–123 (2008)

    Article  CAS  Google Scholar 

  9. Man, P. Y. et al. The epidemiology of Leber hereditary optic neuropathy in the north east of England. Am. J. Hum. Genet. 72, 333–339 (2003)

    Article  CAS  Google Scholar 

  10. Taylor, R. W. & Turnbull, D. M. Mitochondrial DNA mutations in human disease. Nature Rev. Genet. 6, 389–402 (2005)

    Article  CAS  Google Scholar 

  11. Steffann, J. et al. Analysis of mtDNA variant segregation during early human embryonic development: a tool for successful NARP preimplantation diagnosis. J. Med. Genet. 43, 244–247 (2006)

    Article  CAS  Google Scholar 

  12. Piko, L. & Taylor, K. D. Amounts of mitochondrial DNA and abundance of some mitochondrial gene transcripts in early mouse embryos. Dev. Biol. 123, 364–374 (1987)

    Article  CAS  Google Scholar 

  13. Barritt, J. A., Kokot, M., Cohen, J., Steuerwald, N. & Brenner, C. A. Quantification of human ooplasmic mitochondria. Reprod. Biomed. Online 4, 243–247 (2002)

    Article  CAS  Google Scholar 

  14. Lin, D. P. et al. Comparison of mitochondrial DNA contents in human embryos with good or poor morphology at the 8-cell stage. Fertil. Steril. 81, 73–79 (2004)

    Article  CAS  Google Scholar 

  15. Tajima, H. et al. The development of novel quantification assay for mitochondrial DNA heteroplasmy aimed at preimplantation genetic diagnosis of Leigh encephalopathy. J. Assist. Reprod. Genet. 24, 227–232 (2007)

    Article  Google Scholar 

  16. Tachibana, M. et al. Mitochondrial gene replacement in primate offspring and embryonic stem cells. Nature 461, 367–372 (2009)

    Article  ADS  CAS  Google Scholar 

  17. Battaglia, D. E., Goodwin, P., Klein, N. A. & Soules, M. R. Fertilization and early embryology: influence of maternal age on meiotic spindle assembly oocytes from naturally cycling women. Hum. Reprod. 11, 2217–2222 (1996)

    Article  CAS  Google Scholar 

  18. Almeida, P. A. & Bolton, V. N. The effect of temperature fluctuations on the cytoskeletal organisation and chromosomal constitution of the human oocyte. Zygote 3, 357–365 (1995)

    Article  CAS  Google Scholar 

  19. Meirelles, F. V. & Smith, L. C. Mitochondrial genotype segregation in a mouse heteroplasmic lineage produced by embryonic karyoplast transplantation. Genetics 145, 445–451 (1997)

    CAS  PubMed  PubMed Central  Google Scholar 

  20. Feenan, K. & Herbert, M. Can ‘abnormally’ fertilized zygotes give rise to viable embryos? Hum. Fertil. (Camb.) 9, 157–169 (2006)

    Article  Google Scholar 

  21. Chinnery, P., Majamaa, K., Turnbull, D. & Thorburn, D. Treatment for mitochondrial disorders. Cochrane Database Syst. Rev. CD004426 (2006)

  22. McFarland, R. et al. Multiple neonatal deaths due to a homoplasmic mitochondrial DNA mutation. Nature Genet. 30, 145–146 (2002)

    Article  CAS  Google Scholar 

  23. Rahman, S., Poulton, J., Marchington, D. & Suomalainen, A. Decrease of 3243 A→G mtDNA mutation from blood in MELAS syndrome: a longitudinal study. Am. J. Hum. Genet. 68, 238–240 (2001)

    Article  CAS  Google Scholar 

  24. Reik, W. et al. Adult phenotype in the mouse can be affected by epigenetic events in the early embryo. Development 119, 933–942 (1993)

    CAS  PubMed  Google Scholar 

  25. Taylor, R. W., Taylor, G. A., Durham, S. E. & Turnbull, D. M. The determination of complete human mitochondrial DNA sequences in single cells: implications for the study of somatic mitochondrial DNA point mutations. Nucleic Acids Res. 29, e74 (2001)

    Article  CAS  Google Scholar 

  26. Andrews, R. M. et al. Reanalysis and revision of the Cambridge reference sequence for human mitochondrial DNA. Nature Genet. 23, 147 (1999)

    Article  CAS  Google Scholar 

  27. Taylor, R. W. et al. A homoplasmic mitochondrial transfer ribonucleic acid mutation as a cause of maternally inherited hypertrophic cardiomyopathy. J. Am. Coll. Cardiol. 41, 1786–1796 (2003)

    Article  CAS  Google Scholar 

  28. McFarland, R. et al. Familial myopathy: new insights into the T14709C mitochondrial tRNA mutation. Ann. Neurol. 55, 478–484 (2004)

    Article  CAS  Google Scholar 

  29. He, L. et al. Detection and quantification of mitochondrial DNA deletions in individual cells by real-time PCR. Nucleic Acids Res. 30, e68 (2002)

    Article  Google Scholar 

  30. Krishnan, K. J., Bender, A., Taylor, R. W. & Turnbull, D. M. A multiplex real-time PCR method to detect and quantify mitochondrial DNA deletions in individual cells. Anal. Biochem. 370, 127–129 (2007)

    Article  CAS  Google Scholar 

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Acknowledgements

We thank M. Nesbitt, L. Burgess and S. Byerley for help with embryo donation and collection, and V. Wilson and S. Abbs for help with the nuclear genotyping. We thank the patients and staff at Newcastle Fertility Centre, and J. Lawford-Davies, K. Stern, Sir John Burn and Lord Walton of Detchant for helping us to obtain a Human Fertilisation and Embryology Authority research licence and guidance with the legislation. This work was funded by the Muscular Dystrophy Campaign, the Wellcome Trust (074454/Z/04/Z), the Medical Research Council (G0601157, G0601943), One North East, the UK National Institute for Health Research Biomedical Research Centre for Ageing and Age-related Disease and the Newcastle University Centre for Brain Ageing and Vitality supported by the Biotechnology and Biological Sciences Research Council, the Engineering and Physical Sciences Research Council, the Economic and Social Research Council and the Medical Research Council (G0700718). P.F.C. is a Wellcome Trust Senior Fellow in Clinical Science.

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Authors and Affiliations

  1. Mitochondrial Research Group, Institute for Ageing and Health,,

    Lyndsey Craven, Helen A. Tuppen, Julie L. Murphy, Lynsey M. Cree, Patrick F. Chinnery, Robert W. Taylor, Robert N. Lightowlers & Douglass M. Turnbull

  2. Newcastle University Centre for Brain Ageing and Vitality, Institute for Ageing and Health, Newcastle University, Newcastle upon Tyne NE2 4HH, UK,

    Douglass M. Turnbull

  3. Newcastle Fertility Centre, International Centre for Life,,

    Gareth D. Greggains, Stephen J. Harbottle, Alison P. Murdoch & Mary Herbert

  4. Institute for Ageing and Health, International Centre for Life,,

    Gareth D. Greggains & Mary Herbert

  5. North East England Stem Cell Institute (NESCI), Bioscience Centre, International Centre for Life, Newcastle University, Newcastle upon Tyne NE1 4EP, UK,

    Alison P. Murdoch, Mary Herbert & Douglass M. Turnbull

Authors
  1. Lyndsey Craven
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  8. Patrick F. Chinnery
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  11. Mary Herbert
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Contributions

M.H., A.P.M., R.N.L. and D.M.T. conceived the project and designed the experiments. L.C., H.A.T., S.J.H., G.D.G., J.L.M., L.M.C., P.F.C. and R.W.T. performed experiments and analysed data. L.C., M.H., H.A.T. and D.M.T. wrote the manuscript.

Corresponding authors

Correspondence to Mary Herbert or Douglass M. Turnbull.

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The authors declare no competing financial interests.

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Craven, L., Tuppen, H., Greggains, G. et al. Pronuclear transfer in human embryos to prevent transmission of mitochondrial DNA disease. Nature 465, 82–85 (2010). https://doi.org/10.1038/nature08958

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