Metabolic remodeling in hiPSC-derived myofibers carrying the m.3243A>G mutation
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Gabriel E Valdebenito Anitta R Chacko Chih-Yao Chung Preethi Sheshadri Haoyu Chi Benjamin O'Callaghan Monika J Madej Henry Houlden Hannah Rouse Valle Morales Katiuscia Bianchi Francesco Saverio Tedesco Robert DS Pitceathly Michael R DuchenAbstract
Mutations in mitochondrial DNA cause severe multisystem disease frequently associated with muscle weakness. The m.3243A>G mutation is the major cause of mitochondrial encephalomyopathy lactic acidosis and stroke-like episodes (MELAS). Experimental models that recapitulate the disease phenotype in vitro for disease modeling or drug screening are very limited. We have therefore generated hiPSC-derived muscle fibers with variable heteroplasmic mtDNA mutation load without significantly affecting muscle differentiation potential. The cells exhibit physiological characteristics of muscle fibers and show a well-organized myofibrillar structure. In cells carrying the m.3243A>G mutation, the mitochondrial membrane potential and oxygen consumption were reduced in relation to the mutant load. We have shown through proteomic, phosphoproteomic, and metabolomic analyses that the m.3243A>G mutation variably affects the cell phenotype in relation to the mutant load. This variation is reflected by an increase in the NADH/NAD+ ratio, which in turn influences key nutrient-sensing pathways in the myofibers. This model enables a detailed study of the impact of the mutation on cellular bioenergetics and on muscle physiology with the potential to provide a platform for drug screening.
Journal details
Journal
Stem Cell Reports
Volume
20
Issue number
4
Pages
102448
Available online
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10.1016/j.stemcr.2025.102448
Europe PubMed Central
40086445
Pubmed
40086445
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The publication was previously a preprint.
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