Loss of tafazzin results in decreased myoblast differentiation in C2C12 cells: A myoblast model of Barth syndrome and cardiolipin deficiency

Lou, Wenjia; Reynolds, Christian A.; Li, Yiran; Liu, Jenney; Hüttemann, Maik; Schlame, Michael; Stevenson, David; Strathdee, Douglas; Greenberg, Miriam L.

doi:10.1016/j.bbalip.2018.04.015

Cited by 37 publications

(48 citation statements)

References 57 publications

(44 reference statements)

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“…Another common feature observed in multiple cellular models of BTHS is increased oxidative stress . TAZ‐KO in mouse C2C12 myoblasts displayed mitochondrial defects consistent with other models of BTHS and was associated with impairment of myocyte differentiation to myotubes, which may explain the skeletal myopathy observed in BTHS patients .…”

Section: Bths Modelssupporting

confidence: 56%

“…Taz1 deficient yeast cells do not exhibit enhanced sensitivity to various oxidative insults, suggesting that the likely cause for oxidative damage is increased ROS production and not decreased ROS quenching mechanisms . A similar increase in mitochondrial ROS levels was also observed in mammalian cellular BTHS models . Thus, increased ROS is one of the biochemical hallmarks of BTHS.…”

Section: Mitochondrial Dysfunctions In Bthsmentioning

confidence: 72%

“…Many mammalian cellular models are available for BTHS including patient‐derived lymphoblasts , fibroblasts , and induced‐pluripotent stem cells (iPSCs) . Recently, a CRISPR‐mediated stable TAZ‐KO mouse C2C12 myoblast cell line has also been generated . Abnormal mitochondrial morphology is observed in patient‐derived fibroblasts and BTHS iPSC cardiomyocytes .…”

Section: Bths Modelsmentioning

confidence: 99%

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Mitochondrial dysfunctions in barth syndrome

et al. 2019

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Barth syndrome (BTHS) is a rare multisystemic genetic disorder caused by mutations in the TAZ gene. TAZ encodes a mitochondrial enzyme that remodels the acyl chain composition of newly synthesized cardiolipin, a phospholipid unique to mitochondrial membranes. The clinical abnormalities observed in BTHS patients are caused by perturbations in various mitochondrial functions that rely on remodeled cardiolipin. However, the contribution of different cardiolipin‐dependent mitochondrial functions to the pathology of BTHS is not fully understood. In this review, we will discuss recent findings from different genetic models of BTHS, including the yeast model of cardiolipin deficiency that has uncovered the specific in vivo roles of cardiolipin in mitochondrial respiratory chain biogenesis, bioenergetics, intermediary metabolism, mitochondrial dynamics, and quality control. We will also describe findings from higher eukaryotic models of BTHS that highlight a link between cardiolipin‐dependent mitochondrial function and its impact on tissue and organ function. © 2019 IUBMB Life, 9999(9999):1–11, 2019

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Section: Bths Modelssupporting

confidence: 56%

Section: Mitochondrial Dysfunctions In Bthsmentioning

confidence: 72%

Section: Bths Modelsmentioning

confidence: 99%

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Mitochondrial dysfunctions in barth syndrome

et al. 2019

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“…The TAZ-KO cell line used in this study was generated previously using CRISPR/Cas9 targeted against exon 3 of the tafazzin (Taz) gene in C2C12 mouse myoblasts (46). The top 10 predicted off-target sites were identified using the CCTop CRISPR/Cas9 target predictor tool (https://crispr.cos.uniheidelberg.de/) 9 based on the Mus musculus GRCm38 refer-ence genome and the gRNA sequence (5Ј-TCCTAAAACTCC-GCCACATC-3Ј) utilized to create the TAZ-KO cell line (46,63). Next-generation whole-genome sequencing was performed on WT and TAZ-KO cells, and a differential variant analysis was run to identify deviations between the WT and TAZ-KO genomes (GENEWIZ Biotechnology, South Plainfield, NJ).…”

Section: Crispr/cas9 Off-target Analysis Of Taz-ko Cellsmentioning

confidence: 99%

“…To gain insight into the role of CL in energy metabolism, we analyzed the metabolic flux of [U- 13 C]glucose in a tafazzin knockout mouse cell line, TAZ-KO (46). In this study, we demonstrate that the TCA cycle is perturbed in tafazzin-deficient cells, which exhibit decreased flux of glucose carbon to acetyl-CoA, TCA cycle metabolites, and related amino acids.…”

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confidence: 94%

Cardiolipin-induced activation of pyruvate dehydrogenase links mitochondrial lipid biosynthesis to TCA cycle function

Lou

Raja

et al. 2019

Journal of Biological Chemistry

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Edited by George M. Carman Cardiolipin (CL) is the signature phospholipid of mitochondrial membranes. Although it has long been known that CL plays an important role in mitochondrial bioenergetics, recent evidence in the yeast model indicates that CL is also essential for intermediary metabolism. To gain insight into the function of CL in energy metabolism in mammalian cells, here we analyzed the metabolic flux of [U-13 C]glucose in a mouse C2C12 myoblast cell line, TAZ-KO, which is CL-deficient because of CRISPR/Cas9-mediated knockout of the CL-remodeling enzyme tafazzin (TAZ). TAZ-KO cells exhibited decreased flux of [U-13 C]glucose to [ 13 C]acetyl-CoA and M2 and M4 isotopomers of tricarboxylic acid (TCA) cycle intermediates. The activity of pyruvate carboxylase, the predominant enzyme for anaplerotic replenishing of the TCA cycle, was elevated in TAZ-KO cells, which also exhibited increased sensitivity to the pyruvate carboxylase inhibitor phenylacetate. We attributed a decreased carbon flux from glucose to acetyl-CoA in the TAZ-KO cells to a ϳ50% decrease in pyruvate dehydrogenase (PDH) activity, which was observed in both TAZ-KO cells and cardiac tissue from TAZ-KO mice. Protein-lipid overlay experiments revealed that PDH binds to CL, and supplementing digitonin-solubilized TAZ-KO mitochondria with CL restored PDH activity to WT levels. Mitochondria from TAZ-KO cells exhibited an increase in phosphorylated PDH, levels of which were reduced in the presence of supplemented CL. These findings indicate that CL is required for optimal PDH activation, generation of acetyl-CoA, and TCA cycle function, findings that link the key mitochondrial lipid CL to TCA cycle function and energy metabolism. The phospholipid cardiolipin (CL) 8 is the signature lipid of mitochondrial membranes. It interacts with a wide variety of mitochondrial proteins (1) and has multiple roles in mitochondrial biogenesis as well as other cellular functions (2-4). Loss of CL leads to deficiencies in respiratory function, mitochondrial membrane potential, and ATP synthesis (1, 5-7). CL that is synthesized de novo has predominantly saturated fatty acids (8). However, the lipid is then remodeled via phospholipase-mediated deacylation to form monolysocardiolipin (MLCL), which is subsequently acylated by the enzyme tafazzin to synthesize CL with predominantly unsaturated fatty acids (9, 10). Abnormal CL composition is associated with several human disorders, including diabetes, Alzheimer's disease, and Parkinson's disease (11-13). However, Barth syndrome (BTHS) is the only disorder identified to date that is caused by altered CL metabolism (14, 15). BTHS is a severe X-linked disorder characterized by dilated cardiomyopathy, skeletal myopathy, neutropenia, exercise intolerance, lactic acidosis, and sudden death from arrhythmia (16-18). The underlying cause of BTHS is mutations in the tafazzin (TAZ) gene, which codes for the transacylase that remodels CL (10, 19). As a result, BTHS patients exhibit an aberrant CL profile characterized by decreased tot...

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Cardiolipin deficiency in Barth syndrome is not associated with increased superoxide/H₂O₂ production in heart and skeletal muscle mitochondria

Gonçalves¹,

Schlame

Bartelt³

et al. 2020

FEBS Letters

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Barth syndrome (BTHS) is a rare X-linked genetic disorder caused by mutations in the gene encoding the transacylase tafazzin and characterized by loss of cardiolipin and severe cardiomyopathy. Mitochondrial oxidants have been implicated in the cardiomyopathy in BTHS. Eleven mitochondrial sites produce superoxide/hydrogen peroxide (H 2 O 2) at significant rates. Which of these sites generate oxidants at excessive rates in BTHS is unknown. Here, we measured the maximum capacity of superoxide/H 2 O 2 production from each site and the ex vivo rate of superoxide/H 2 O 2 production in the heart and skeletal muscle mitochondria of the tafazzin knockdown mice (tazkd) from 3 to 12 months of age. Despite reduced oxidative capacity, superoxide/H 2 O 2 production was indistinguishable between tazkd mice and wild-type littermates. These observations raise questions about the involvement of mitochondrial oxidants in BTHS pathology.

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Loss of tafazzin results in decreased myoblast differentiation in C2C12 cells: A myoblast model of Barth syndrome and cardiolipin deficiency

Cited by 37 publications

References 57 publications

Mitochondrial dysfunctions in barth syndrome

Mitochondrial dysfunctions in barth syndrome

Cardiolipin-induced activation of pyruvate dehydrogenase links mitochondrial lipid biosynthesis to TCA cycle function

Cardiolipin deficiency in Barth syndrome is not associated with increased superoxide/H₂O₂ production in heart and skeletal muscle mitochondria

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Loss of tafazzin results in decreased myoblast differentiation in C2C12 cells: A myoblast model of Barth syndrome and cardiolipin deficiency

Cited by 37 publications

References 57 publications

Mitochondrial dysfunctions in barth syndrome

Mitochondrial dysfunctions in barth syndrome

Cardiolipin-induced activation of pyruvate dehydrogenase links mitochondrial lipid biosynthesis to TCA cycle function

Cardiolipin deficiency in Barth syndrome is not associated with increased superoxide/H2O2 production in heart and skeletal muscle mitochondria

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Cardiolipin deficiency in Barth syndrome is not associated with increased superoxide/H₂O₂ production in heart and skeletal muscle mitochondria