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 Amato; Don P. Wolf; Taosheng Huang; Andre Terzic; Louise C. Laurent; Juan Carlos Izpisua Belmonte; Shoukhrat Mitalipov

doi:10.1038/nature14546

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

Biological, Environmental Sciences and Engineering

Research output: Contribution to journal › Article › peer-review

152 Scopus citations

Abstract

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 language	English (US)
Pages (from-to)	234-238
Number of pages	5
Journal	Nature
Volume	524
Issue number	7564
DOIs	https://doi.org/10.1038/nature14546
State	Published - 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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General

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This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1038/nature14546

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Ma, H., Folmes, C. D. L., Wu, J., Morey, R., Mora-Castilla, S., Ocampo, A., Ma, L., Poulton, J., Wang, X., Ahmed, R., Kang, E., Lee, Y., Hayama, T., Li, Y., Van Dyken, C., Gutierrez, N. M., Tippner-Hedges, R., Koski, A., Mitalipov, N., ... Mitalipov, S. (2015). Metabolic rescue in pluripotent cells from patients with mtDNA disease. Nature, 524(7564), 234-238. https://doi.org/10.1038/nature14546

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title = "Metabolic rescue in pluripotent cells from patients with mtDNA disease",

abstract = "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.",

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

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{\textquoteright}s Health Research Unit staff at the Center for Women{\textquoteright}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: {\textcopyright} 2015 Macmillan Publishers Limited. All rights reserved.",

year = "2015",

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doi = "10.1038/nature14546",

language = "English (US)",

volume = "524",

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journal = "Nature",

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Ma, H, Folmes, CDL, Wu, J, Morey, R, Mora-Castilla, S, Ocampo, A, Ma, L, Poulton, J, Wang, X, Ahmed, R, Kang, E, Lee, Y, Hayama, T, Li, Y, Van Dyken, C, Gutierrez, NM, Tippner-Hedges, R, Koski, A, Mitalipov, N, Amato, P, Wolf, DP, Huang, T, Terzic, A, Laurent, LC, Belmonte, JCI & Mitalipov, S 2015, 'Metabolic rescue in pluripotent cells from patients with mtDNA disease', Nature, vol. 524, no. 7564, pp. 234-238. https://doi.org/10.1038/nature14546

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T1 - Metabolic rescue in pluripotent cells from patients with mtDNA disease

AU - Ma, Hong

AU - Folmes, Clifford D.L.

AU - Wu, Jun

AU - Morey, Robert

AU - Mora-Castilla, Sergio

AU - Ocampo, Alejandro

AU - Ma, Li

AU - Poulton, Joanna

AU - Wang, Xinjian

AU - Ahmed, Riffat

AU - Kang, Eunju

AU - Lee, Yeonmi

AU - Hayama, Tomonari

AU - Li, Ying

AU - Van Dyken, Crystal

AU - Gutierrez, Nuria Marti

AU - Tippner-Hedges, Rebecca

AU - Koski, Amy

AU - Mitalipov, Nargiz

AU - Amato, Paula

AU - Wolf, Don P.

AU - Huang, Taosheng

AU - Terzic, Andre

AU - Laurent, Louise C.

AU - Belmonte, Juan Carlos Izpisua

AU - Mitalipov, Shoukhrat

N1 - 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.

PY - 2015/8/13

Y1 - 2015/8/13

N2 - 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.

AB - 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.

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JO - Nature

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Metabolic rescue in pluripotent cells from patients with mtDNA disease

Abstract

Bibliographical note

ASJC Scopus subject areas

UN SDGs

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