Disease-causing mitochondrial Heteroplasmy segregated within induced pluripotent stem cell clones derived from a patient with MELAS

Clifford Folmes, Almudena Martinez-Fernandez, Ester Perales-Clemente, Xing Li, Amber McDonald, Devin Oglesbee, Sybil C. Hrstka, Carmen M Terzic, Andre Terzic, Timothy J Nelson

Research output: Contribution to journalArticle

73 Citations (Scopus)

Abstract

Mitochondrial diseases display pathological phenotypes according to the mixture of mutant versus wild-type mitochondrial DNA (mtDNA), known as heteroplasmy. We herein examined the impact of nuclear reprogramming and clonal isolation of induced pluripotent stem cells (iPSC) on mitochondrial heteroplasmy. Patient-derived dermal fibroblasts with a prototypical mitochondrial deficiency diagnosed as mitochondrial encephalomyopathy with lactic acidosis and stroke-like episodes (MELAS) demonstrated mitochondrial dysfunction with reduced oxidative reserve due to heteroplasmy at position G13513A in the ND5 subunit of complex I. Bioengineered iPSC clones acquired pluripotency with multilineage differentiation capacity and demonstrated reduction in mitochondrial density and oxygen consumption distinguishing them from the somatic source. Consistent with the cellular mosaicism of the original patient-derived fibroblasts, the MELAS-iPSC clones contained a similar range of mtDNA heteroplasmy of the disease-causing mutation with identical profiles in the remaining mtDNA. High-heteroplasmy iPSC clones were used to demonstrate that extended stem cell passaging was sufficient to purge mutant mtDNA, resulting in isogenic iPSC subclones with various degrees of disease-causing genotypes. On comparative differentiation of iPSC clones, improved cardiogenic yield was associated with iPSC clones containing lower heteroplasmy compared with isogenic clones with high heteroplasmy. Thus, mtDNA heteroplasmic segregation within patientderived stem cell lines enables direct comparison of genotype/phenotype relationships in progenitor cells and lineage-restricted progeny, and indicates that cell fate decisions are regulated as a function of mtDNA mutation load. The novel nuclear reprogramming-based model system introduces a disease-in-a-dish tool to examine the impact of mutant genotypes for MELAS patients in bioengineered tissues and a cellular probe for molecular features of individual mitochondrial diseases.

Original languageEnglish (US)
Pages (from-to)1298-1308
Number of pages11
JournalStem Cells
Volume31
Issue number7
DOIs
StatePublished - Jul 2013

Fingerprint

MELAS Syndrome
Induced Pluripotent Stem Cells
Mitochondrial Diseases
Mitochondrial DNA
Clone Cells
Stem Cells
Genotype
Fibroblasts
Phenotype
Molecular Probes
Mutation
Mosaicism
Cell Lineage
Oxygen Consumption
Cell Line
Skin

Keywords

  • Induced pluripotent stem cells
  • MELAS syndrome
  • Mitochondria
  • Mitochondrial DNA
  • Regenerative medicine

ASJC Scopus subject areas

  • Cell Biology
  • Developmental Biology
  • Molecular Medicine

Cite this

Disease-causing mitochondrial Heteroplasmy segregated within induced pluripotent stem cell clones derived from a patient with MELAS. / Folmes, Clifford; Martinez-Fernandez, Almudena; Perales-Clemente, Ester; Li, Xing; McDonald, Amber; Oglesbee, Devin; Hrstka, Sybil C.; Terzic, Carmen M; Terzic, Andre; Nelson, Timothy J.

In: Stem Cells, Vol. 31, No. 7, 07.2013, p. 1298-1308.

Research output: Contribution to journalArticle

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abstract = "Mitochondrial diseases display pathological phenotypes according to the mixture of mutant versus wild-type mitochondrial DNA (mtDNA), known as heteroplasmy. We herein examined the impact of nuclear reprogramming and clonal isolation of induced pluripotent stem cells (iPSC) on mitochondrial heteroplasmy. Patient-derived dermal fibroblasts with a prototypical mitochondrial deficiency diagnosed as mitochondrial encephalomyopathy with lactic acidosis and stroke-like episodes (MELAS) demonstrated mitochondrial dysfunction with reduced oxidative reserve due to heteroplasmy at position G13513A in the ND5 subunit of complex I. Bioengineered iPSC clones acquired pluripotency with multilineage differentiation capacity and demonstrated reduction in mitochondrial density and oxygen consumption distinguishing them from the somatic source. Consistent with the cellular mosaicism of the original patient-derived fibroblasts, the MELAS-iPSC clones contained a similar range of mtDNA heteroplasmy of the disease-causing mutation with identical profiles in the remaining mtDNA. High-heteroplasmy iPSC clones were used to demonstrate that extended stem cell passaging was sufficient to purge mutant mtDNA, resulting in isogenic iPSC subclones with various degrees of disease-causing genotypes. On comparative differentiation of iPSC clones, improved cardiogenic yield was associated with iPSC clones containing lower heteroplasmy compared with isogenic clones with high heteroplasmy. Thus, mtDNA heteroplasmic segregation within patientderived stem cell lines enables direct comparison of genotype/phenotype relationships in progenitor cells and lineage-restricted progeny, and indicates that cell fate decisions are regulated as a function of mtDNA mutation load. The novel nuclear reprogramming-based model system introduces a disease-in-a-dish tool to examine the impact of mutant genotypes for MELAS patients in bioengineered tissues and a cellular probe for molecular features of individual mitochondrial diseases.",
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AU - Folmes, Clifford

AU - Martinez-Fernandez, Almudena

AU - Perales-Clemente, Ester

AU - Li, Xing

AU - McDonald, Amber

AU - Oglesbee, Devin

AU - Hrstka, Sybil C.

AU - Terzic, Carmen M

AU - Terzic, Andre

AU - Nelson, Timothy J

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