Induced pluripotent stem cells derived from patients carrying mitochondrial mutations exhibit altered bioenergetics and aberrant differentiation potential.
Autor: | Meshrkey F; Department of Biological Sciences, J. William Fulbright College of Arts and Sciences, University of Arkansas, Science and Engineering 601, Fayetteville, AR, 72701, USA.; Cell and Molecular Biology Program, University of Arkansas, Fayetteville, AR, USA.; Department of Histology and Cell Biology, Faculty of Medicine, Alexandria University, Alexandria, Egypt., Scheulin KM; Regenerative Bioscience Center, University of Georgia, Athens, GA, USA.; Department of Animal and Dairy Science, University of Georgia, Athens, GA, USA.; Neuroscience Program, Biomedical and Health Sciences Institute, University of Georgia, Athens, GA, USA., Littlejohn CM; Regenerative Bioscience Center, University of Georgia, Athens, GA, USA.; Department of Animal and Dairy Science, University of Georgia, Athens, GA, USA., Stabach J; Department of Biological Sciences, J. William Fulbright College of Arts and Sciences, University of Arkansas, Science and Engineering 601, Fayetteville, AR, 72701, USA., Saikia B; Department of Biological Sciences, J. William Fulbright College of Arts and Sciences, University of Arkansas, Science and Engineering 601, Fayetteville, AR, 72701, USA., Thorat V; Department of Genetics and Genome Sciences, Case Western Reserve University School of Medicine, Cleveland, OH, USA., Huang Y; Department of Genetics and Genome Sciences, Case Western Reserve University School of Medicine, Cleveland, OH, USA., LaFramboise T; Department of Genetics and Genome Sciences, Case Western Reserve University School of Medicine, Cleveland, OH, USA., Lesnefsky EJ; Department of Physiology and Biophysics, Virginia Commonwealth University, Richmond, VA, USA.; Cardiology Section Medical Service, McGuire Veterans Affairs Medical Center, Richmond, VA, USA.; Department of Biochemistry and Molecular Biology, Virginia Commonwealth University, Richmond, VA, USA.; Division of Cardiology, Department of Internal Medicine, Pauley Heart Center, Virginia Commonwealth University, Richmond, VA, USA., Rao RR; Department of Biomedical Engineering, College of Engineering, University of Arkansas, Fayetteville, AR, USA., West FD; Regenerative Bioscience Center, University of Georgia, Athens, GA, USA.; Department of Animal and Dairy Science, University of Georgia, Athens, GA, USA.; Neuroscience Program, Biomedical and Health Sciences Institute, University of Georgia, Athens, GA, USA., Iyer S; Department of Biological Sciences, J. William Fulbright College of Arts and Sciences, University of Arkansas, Science and Engineering 601, Fayetteville, AR, 72701, USA. si012@uark.edu.; Cell and Molecular Biology Program, University of Arkansas, Fayetteville, AR, USA. si012@uark.edu. |
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Jazyk: | angličtina |
Zdroj: | Stem cell research & therapy [Stem Cell Res Ther] 2023 Nov 07; Vol. 14 (1), pp. 320. Date of Electronic Publication: 2023 Nov 07. |
DOI: | 10.1186/s13287-023-03546-7 |
Abstrakt: | Background: Human mitochondrial DNA mutations are associated with common to rare mitochondrial disorders, which are multisystemic with complex clinical pathologies. The pathologies of these diseases are poorly understood and have no FDA-approved treatments leading to symptom management. Leigh syndrome (LS) is a pediatric mitochondrial disorder that affects the central nervous system during early development and causes death in infancy. Since there are no adequate models for understanding the rapid fatality associated with LS, human-induced pluripotent stem cell (hiPSC) technology has been recognized as a useful approach to generate patient-specific stem cells for disease modeling and understanding the origins of the phenotype. Methods: hiPSCs were generated from control BJ and four disease fibroblast lines using a cocktail of non-modified reprogramming and immune evasion mRNAs and microRNAs. Expression of hiPSC-associated intracellular and cell surface markers was identified by immunofluorescence and flow cytometry. Karyotyping of hiPSCs was performed with cytogenetic analysis. Sanger and next-generation sequencing were used to detect and quantify the mutation in all hiPSCs. The mitochondrial respiration ability and glycolytic function were measured by the Seahorse Bioscience XFe96 extracellular flux analyzer. Results: Reprogrammed hiPSCs expressed pluripotent stem cell markers including transcription factors POU5F1, NANOG and SOX2 and cell surface markers SSEA4, TRA-1-60 and TRA-1-81 at the protein level. Sanger sequencing analysis confirmed the presence of mutations in all reprogrammed hiPSCs. Next-generation sequencing demonstrated the variable presence of mutant mtDNA in reprogrammed hiPSCs. Cytogenetic analyses confirmed the presence of normal karyotype in all reprogrammed hiPSCs. Patient-derived hiPSCs demonstrated decreased maximal mitochondrial respiration, while mitochondrial ATP production was not significantly different between the control and disease hiPSCs. In line with low maximal respiration, the spare respiratory capacity was lower in all the disease hiPSCs. The hiPSCs also demonstrated neural and cardiac differentiation potential. Conclusion: Overall, the hiPSCs exhibited variable mitochondrial dysfunction that may alter their differentiation potential and provide key insights into clinically relevant developmental perturbations. (© 2023. The Author(s).) |
Databáze: | MEDLINE |
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