Towards Clinical Application of Pronuclear Transfer to Prevent Mitochondrial DNA Disease

Nature. 2016 Jun 16;534(7607):383-6. doi: 10.1038/nature18303. Epub 2016 Jun 8.

Abstract

Mitochondrial DNA (mtDNA) mutations are maternally inherited and are associated with a broad range of debilitating and fatal diseases. Reproductive technologies designed to uncouple the inheritance of mtDNA from nuclear DNA may enable affected women to have a genetically related child with a greatly reduced risk of mtDNA disease. Here we report the first preclinical studies on pronuclear transplantation (PNT). Surprisingly, techniques used in proof-of-concept studies involving abnormally fertilized human zygotes were not well tolerated by normally fertilized zygotes. We have therefore developed an alternative approach based on transplanting pronuclei shortly after completion of meiosis rather than shortly before the first mitotic division. This promotes efficient development to the blastocyst stage with no detectable effect on aneuploidy or gene expression. After optimization, mtDNA carryover was reduced to <2% in the majority (79%) of PNT blastocysts. The importance of reducing carryover to the lowest possible levels is highlighted by a progressive increase in heteroplasmy in a stem cell line derived from a PNT blastocyst with 4% mtDNA carryover. We conclude that PNT has the potential to reduce the risk of mtDNA disease, but it may not guarantee prevention.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Blastocyst / cytology
  • Blastocyst / metabolism
  • Cell Nucleus / genetics
  • Cell Nucleus / metabolism
  • Cytoplasm / genetics
  • Cytoplasm / metabolism
  • DNA, Mitochondrial / analysis
  • DNA, Mitochondrial / genetics*
  • Female
  • Gene Expression Profiling
  • Humans
  • Male
  • Meiosis
  • Mitochondria / genetics
  • Mitochondria / metabolism
  • Mitochondrial Diseases / genetics*
  • Mitochondrial Diseases / pathology
  • Mitochondrial Diseases / prevention & control*
  • Mitochondrial Replacement Therapy / methods*
  • Nuclear Transfer Techniques*
  • Stem Cells / cytology
  • Stem Cells / metabolism
  • Translational Medical Research
  • Young Adult
  • Zygote / cytology
  • Zygote / metabolism

Substances

  • DNA, Mitochondrial