Metabolic rescue in pluripotent cells from patients with mtDNA disease

Hong Ma, Clifford D.L. Folmes, Jun Wu, Robert Morey, Sergio Mora-Castilla, Alejandro Ocampo, Li Ma, Joanna Poulton, Xinjian Wang, Riffat Ahmed, Eunju Kang, Yeonmi Lee, Tomonari Hayama, Ying Li, Crystal Van Dyken, Nuria Marti Gutierrez, Rebecca Tippner-Hedges, Amy Koski, Nargiz Mitalipov, Paula AmatoDon P. Wolf, Taosheng Huang, Andre Terzic, Louise C. Laurent, Juan Carlos Izpisua Belmonte, Shoukhrat Mitalipov*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

152 Scopus citations


Mitochondria have a major role in energy production via oxidative phosphorylation, which is dependent on the expression of critical genes encoded by mitochondrial (mt)DNA. Mutations in mtDNA can cause fatal or severely debilitating disorders with limited treatment options. Clinical manifestations vary based on mutation type and heteroplasmy (that is, the relative levels of mutant and wild-type mtDNA within each cell). Here we generated genetically corrected pluripotent stem cells (PSCs) from patients with mtDNA disease. Multiple induced pluripotent stem (iPS) cell lines were derived from patients with common heteroplasmic mutations including 3243A>G, causing mitochondrial encephalomyopathy and stroke-like episodes (MELAS), and 8993T>G and 13513G>A, implicated in Leigh syndrome. Isogenic MELAS and Leigh syndrome iPS cell lines were generated containing exclusively wild-type or mutant mtDNA through spontaneous segregation of heteroplasmic mtDNA in proliferating fibroblasts. Furthermore, somatic cell nuclear transfer (SCNT) enabled replacement of mutant mtDNA from homoplasmic 8993T>G fibroblasts to generate corrected Leigh-NT1 PSCs. Although Leigh-NT1 PSCs contained donor oocyte wild-type mtDNA (human haplotype D4a) that differed from Leigh syndrome patient haplotype (F1a) at a total of 47 nucleotide sites, Leigh-NT1 cells displayed transcriptomic profiles similar to those in embryo-derived PSCs carrying wild-type mtDNA, indicative of normal nuclear-to-mitochondrial interactions. Moreover, genetically rescued patient PSCs displayed normal metabolic function compared to impaired oxygen consumption and ATP production observed in mutant cells. We conclude that both reprogramming approaches offer complementary strategies for derivation of PSCs containing exclusively wild-type mtDNA, through spontaneous segregation of heteroplasmic mtDNA in individual iPS cell lines or mitochondrial replacement by SCNT in homoplasmic mtDNA-based disease.

Original languageEnglish (US)
Pages (from-to)234-238
Number of pages5
Issue number7564
StatePublished - Aug 13 2015

Bibliographical note

Funding Information:
Acknowledgements The authors acknowledge the OHSU Embryonic Stem Cell Research Oversight Committee and the Institutional Review Board for providing oversight and guidance. We thank skin and oocyte donors and the Women’s Health Research Unit staff at the Center for Women’s Health, University Fertility Consultants and the Reproductive Endocrinology & Infertility Division in the Department of Obstetrics & Gynecology of Oregon Health & Science University for their support and procurement of gametes. We are grateful to M. Tachibana and A. Polat for help with derivation of PSCs and to M. Sparman for technical support. We are indebted to S. Gokhale for teratoma analysis and M. C. T. Penedo for microsatellite genotyping. We thank the staff at the Institute for Genomic Medicine Genomics Facility at UCSD for sequencing the RNA-seq libraries. Studies were supported by the Leducq Foundation, Mayo Clinic Center for Regenerative Medicine andOHSU and UCSD institutionalfunds. Work in the laboratory of J.C.I.B. was supported by the G. Harold and Leila Y. Mathers Charitable Foundation and the Leona M. and Harry B. Helmsley Charitable Trust (2012-PG-MED002).

Publisher Copyright:
© 2015 Macmillan Publishers Limited. All rights reserved.

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