Dataset Information

Metabolic remodelling in hiPSC-derived myofibres carrying the m.3243A>G mutation

ABSTRACT: 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 modelling or drug screening are very limited. We have therefore generated hiPSC-derived muscle fibres with variable heteroplasmic mtDNA mutation load without significantly affecting muscle differentiation potential. The cells are excitable and show physiological characteristics of muscle fibres and show well organised myofibrillar structure. In cells carrying the m.3243A>G, 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 turns influences key nutrient-sensing pathways in the myofibres. This model enables 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.

INSTRUMENT(S): Orbitrap Eclipse

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: riccardo zenezini chiozzi

LAB HEAD: Konstantinos Thalassinos

PROVIDER: PXD058785 | Pride | 2025-05-26

REPOSITORIES: Pride

ACCESS DATA

Dataset's files

Source:

			Action	DRS
	20241210_114942_STP0071_myoblast_proteome.sne	Other
	20241210_141055_STP0071_myofibre_proteome.sne	Other
	20241211_112719_STP0071_myoblast_phos.sne	Other
	20241211_131637_STP0071_myofibre_phos.sne	Other
	30082023_RZC_STP0071_Myoblast_condition0_01.raw	Raw

Items per page:

1 - 5 of 48

Publications

Metabolic remodeling in hiPSC-derived myofibers carrying the m.3243A>G mutation.

Valdebenito Gabriel E GE Chacko Anitta R AR Chung Chih-Yao CY Sheshadri Preethi P Chi Haoyu H O'Callaghan Benjamin B Madej Monika J MJ Houlden Henry H Rouse Hannah H Morales Valle V Bianchi Katiuscia K Tedesco Francesco Saverio FS Pitceathly Robert D S RDS Duchen Michael R MR

Stem cell reports 20250313 4

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 mus ...[more]

PMID: 40086445

Similar Datasets

Project description:Pyruvate oxidase encoded by spxB is a major virulence factor in the human respiratory pathogen Streptococcus pneumoniae. During aerobic growth, SpxB synthesizes large amounts of H2O2 and acetyl phosphate, which can serve as a phosphoryl group donor to response regulators and be converted to ATP. SpxB is the main source of the millimolar concentrations of H2O2 produced and tolerated by pneumococcus, despite its lack of a catalase. We report here the first cis- and trans-acting regulatory elements for spxB transcription. These elements were identified in a genetic screen, similar to those used previously for phase variants, for spontaneous mutations that caused colonies of virulent serotype 2 strain D39 to change from a transparent to an opaque appearance. Six of the seven opaque colonies recovered (frequency of 3 x 10-5) were impaired for SpxB function. Modeling suggested that two mutations changed amino acids in SpxB required for FAD cofactor or subunit binding. One mutation deleted a cis-acting adjacent direct repeat required for optimal spxB transcription. The other three independent mutations created the same frameshift near the start of a trans-acting regulatory gene designated as spxR. The SpxR protein contains helix-turn-helix, CBS, and HotDog domains implicated in DNA, adenosine, and CoA compound binding, respectively, consistent with the idea that SpxR positively regulates spxB transcript amount in response to energy and metabolic state rather than oxidative state. Finally, microarray analyses of a null spxB or a spxR mutant revealed the presence of a new oxidative stress response in pneumococcus and unexpectedly demonstrated that SpxR strongly positively regulates the transcript amount of the strH exoglycosidase gene, which like spxB, has been implicated in host colonization. Keywords: genetic modification Bacterial strains were grown exponentially in rich (BHI) media at 37C and an atmosphere of 5% CO2, and were processed as described in the related Sample records. Samples were collected from three independent biological replicates and included one dye swap. Data were normalized using the Lowess (subgrid) method without background subtraction.

Dataset Information

Metabolic remodelling in hiPSC-derived myofibres carrying the m.3243A>G mutation

Dataset's files

Publications

Metabolic remodeling in hiPSC-derived myofibers carrying the m.3243A>G mutation.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets

Dataset Information

Metabolic remodelling in hiPSC-derived myofibres carrying the m.3243A>G mutation

Dataset's files

Publications

Metabolic remodeling in hiPSC-derived myofibers carrying the m.3243A&gt;G mutation.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets

Metabolic remodeling in hiPSC-derived myofibers carrying the m.3243A>G mutation.