title: Natural underlying mtDNA heteroplasmy as a potential source of intra-person hiPSC variability.
abstract: Functional variability among human clones of induced pluripotent stem cells (hiPSCs) remains a limitation in assembling high-quality biorepositories. Beyond inter-person variability, the root cause of intra-person variability remains unknown. Mitochondria guide the required transition from oxidative to glycolytic metabolism in nuclear reprogramming. Moreover, mitochondria have their own genome (mitochondrial DNA [mtDNA]). Herein, we performed mtDNA next-generation sequencing (NGS) on 84 hiPSC clones derived from a cohort of 19 individuals, including mitochondrial and non-mitochondrial patients. The analysis of mtDNA variants showed that low levels of potentially pathogenic mutations in the original fibroblasts are revealed through nuclear reprogramming, generating mutant hiPSCs with a detrimental effect in their differentiated progeny. Specifically, hiPSC-derived cardiomyocytes with expanded mtDNA mutations non-related with any described human disease, showed impaired mitochondrial respiration, being a potential cause of intra-person hiPSC variability. We propose mtDNA NGS as a new selection criterion to ensure hiPSC quality for drug discovery and regenerative medicine.
authors.affiliation: Departments of Medicine, Molecular Pharmacology and Experimental Therapeutics, and Medical Genetics, Division of Cardiovascular Diseases, Mayo Clinic Center for Regenerative Medicine, Rochester, MN, USA.
authors.forename: Ester
authors.initials: E
authors.lastname: Perales-Clemente
authors.affiliation: Departments of Cardiovascular Diseases, Molecular Pharmacology and Experimental Therapeutics, Division of General Internal Medicine, Division of Pediatric Cardiology, and Transplant Center, Mayo Clinic Center for Regenerative Medicine, Rochester, MN, USA.
authors.forename: Alexandra N
authors.initials: AN
authors.lastname: Cook
authors.affiliation: Division of Biomedical Statistics and Informatics, Department of Health Sciences Research, Mayo Clinic, Rochester, MN, USA.
authors.forename: Jared M
authors.initials: JM
authors.lastname: Evans
authors.affiliation: Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, MN, USA.
authors.forename: Samantha
authors.initials: S
authors.lastname: Roellinger
authors.affiliation: Departments of Cardiovascular Diseases, Molecular Pharmacology and Experimental Therapeutics, Division of General Internal Medicine, Division of Pediatric Cardiology, and Transplant Center, Mayo Clinic Center for Regenerative Medicine, Rochester, MN, USA.
authors.forename: Frank
authors.initials: F
authors.lastname: Secreto
authors.affiliation: Department of Clinical and Experimental Medicine, University of Messina, Messina, Italy.
authors.forename: Valentina
authors.initials: V
authors.lastname: Emmanuele
authors.affiliation: Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, MN, USA.
authors.forename: Devin
authors.initials: D
authors.lastname: Oglesbee
authors.affiliation: Department of Molecular Biology, Howard Hughes Medical Institute Massachusetts General Hospital, Boston, MA, USA.
authors.forename: Vamsi K
authors.initials: VK
authors.lastname: Mootha
authors.affiliation: Department of Neurology, Columbia University Medical Center, New York, NY, USA.
authors.forename: Michio
authors.initials: M
authors.lastname: Hirano
authors.affiliation: Department of Neurology, Columbia University Medical Center, New York, NY, USA Department of Genetics and Development, Columbia University Medical Center, New York, NY, USA.
authors.forename: Eric A
authors.initials: EA
authors.lastname: Schon
authors.affiliation: Departments of Medicine, Molecular Pharmacology and Experimental Therapeutics, and Medical Genetics, Division of Cardiovascular Diseases, Mayo Clinic Center for Regenerative Medicine, Rochester, MN, USA.
authors.forename: Andre
authors.initials: A
authors.lastname: Terzic
authors.affiliation: Departments of Cardiovascular Diseases, Molecular Pharmacology and Experimental Therapeutics, Division of General Internal Medicine, Division of Pediatric Cardiology, and Transplant Center, Mayo Clinic Center for Regenerative Medicine, Rochester, MN, USA nelson.timothy@mayo.edu.
authors.forename: Timothy J
authors.identifier: 0000-0002-3862-7023
authors.initials: TJ
authors.lastname: Nelson
mesh_terms: D002454:Cell Differentiation; D019069:Cell Respiration; D004272:DNA, Mitochondrial / Q000737:chemistry / Q000235:genetics*; D014644:Genetic Variation; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000502:physiology*; D009154:Mutation; D032383:Myocytes, Cardiac / Q000502:physiology; D010641:Phenotype
chemical_list: D004272:DNA, Mitochondrial
keywords: global private mutation; human iPSC; intra‐person variability; mitochondrial DNA; quality control; universal heteroplasmy