The Caveolin-3 P104L mutation of LGMD-1C leads to disordered glucose metabolism in muscle cells

Deng, Yu-Feng; Huang, Yi Yuan; Lu, Wensheng; Huang, Yuan Heng; Xian, Jing; Wei, Hong; Huang, Qin

doi:10.1016/j.bbrc.2017.02.072

Cited by 14 publications

(23 citation statements)

References 29 publications

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“…This restraint is likely to be alleviated in muscle expressing the Cav3 P104L mutant owing to the substantial reduction in Cav3 that would result in a concomitant increase in myostatin signalling . Interestingly, very recent work has also linked expression of Cav3 P104L in C2C12 myotubes to reduced activation of Akt (a key insulin signalling intermediate) and a diminution in glucose uptake and glycogen synthesis . These latter observations raise the possibility that disturbances in fuel and energy metabolism may also be contributing factors to the pathology of LGMD1C.…”

Section: Introductionmentioning

confidence: 99%

Caveolin‐3 deficiency associated with the dystrophy P104L mutation impairs skeletal muscle mitochondrial form and function

Shah

Nisr

Stretton

et al. 2020

J cachexia sarcopenia muscle

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Background Caveolin‐3 (Cav3) is the principal structural component of caveolae in skeletal muscle. Dominant pathogenic mutations in the Cav3 gene, such as the Limb Girdle Muscular Dystrophy‐1C (LGMD1C) P104L mutation, result in substantial loss of Cav3 and myopathic changes characterized by muscle weakness and wasting. We hypothesize such myopathy may also be associated with disturbances in mitochondrial biology. Herein, we report studies assessing the effects of Cav3 deficiency on mitochondrial form and function in skeletal muscle cells. Methods L6 myoblasts were stably transfected with Cav3P104L or expression of native Cav3 repressed by shRNA or CRISPR/Cas9 genome editing prior to performing fixed/live cell imaging of mitochondrial morphology, subcellular fractionation and immunoblotting, or analysis of real time mitochondrial respiration. Skeletal muscle from wild‐type and Cav3−/− mice was processed for analysis of mitochondrial proteins by immunoblotting. Results Caveolin‐3 was detected in mitochondrial‐enriched membranes isolated from mouse gastrocnemius muscle and L6 myoblasts. Expression of Cav3P104L in L6 myoblasts led to its targeting to the Golgi and loss of native Cav3 (>95%), including that associated with mitochondrial membranes. Cav3P104L reduced mitochondrial mass and induced fragmentation of the mitochondrial network that was associated with significant loss of proteins involved in mitochondrial biogenesis, respiration, morphology, and redox function [i.e. PGC1α, succinate dehyrdogenase (SDHA), ANT1, MFN2, OPA1, and MnSOD). Furthermore, Cav3P104L myoblasts exhibited increased mitochondrial cholesterol and loss of cardiolipin. Consistent with these changes, Cav3P104L expression reduced mitochondrial respiratory capacity and increased myocellular superoxide production. These morphological, biochemical, and functional mitochondrial changes were phenocopied in myoblasts in which Cav3 had been silenced/knocked‐out using shRNA or CRISPR. Reduced mitochondrial mass, PGC1α, SDHA, ANT1, and MnSOD were also demonstrable in Cav3−/− mouse gastrocnemius. Strikingly, Cav3 re‐expression in Cav3KO myoblasts restored its mitochondrial association and facilitated reformation of a tubular mitochondrial network. Significantly, re‐expression also mitigated changes in mitochondrial superoxide, cholesterol, and cardiolipin content and recovered cellular respiratory capacity. Conclusions Our results identify Cav3 as an important regulator of mitochondrial homeostasis and reveal that Cav3 deficiency in muscle cells associated with the Cav3P104L mutation invokes major disturbances in mitochondrial respiration and energy status that may contribute to the pathology of LGMD1C.

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Section: Introductionmentioning

confidence: 99%

Caveolin‐3 deficiency associated with the dystrophy P104L mutation impairs skeletal muscle mitochondrial form and function

Shah

Nisr

Stretton

et al. 2020

J cachexia sarcopenia muscle

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Section: P104l Changed Non-insulin-stimulation Glucose Metabolism In mentioning

confidence: 47%

“…Thus, CAV3‐P104L cannot be appropriately localized in the cell membrane, and the total CAV3 content is also reduced. In our previous study, under insulin stimulation, the expression of CAV3 protein was reduced by approximately 90% in cells transfected with the P104L mutant gene compared with the wild‐type gene (Deng et al, ). In this study, the CAV3 level was reduced by approximately 82.7% without insulin stimulation.…”

Section: Discussionmentioning

confidence: 93%

“…Previous studies have shown that the P104L mutation leads to decreased glucose uptake and glycogen synthesis, and glucose metabolism disruption in skeletal muscle cells treated with insulin (Deng et al, ). In this study, normal C2C12 cells were cultured without insulin stimulation and transfected with the P104L mutant gene, which induced abnormal glucose metabolism, such as reduced glucose uptake and decreased glycogen synthesis, and the change was primarily non‐insulin‐stimulation.…”

Section: Discussionmentioning

confidence: 99%

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The caveolin‐3 P104L mutation in LGMD‐1C patients inhibits non‐insulin‐stimulated glucose metabolism and growth but promotes myocyte proliferation

Shang

Chen

Xian

et al. 2019

Cell Biology International

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The caveolin‐3 (CAV3) protein is known to be specifically expressed in various myocytes, and skeletal muscle consumes most of the blood glucose as an energy source to maintain normal cell metabolism and function. The P104L mutation in the coding sequence of the human CAV3 gene leads to autosomal dominant disease limb‐girdle muscular dystrophy type 1C (LGMD‐1C). We previously reported that C2C12 cells transiently transfected with the P104L CAV3 mutant exhibited decreased glucose uptake and glycogen synthesis after insulin stimulation. The present study aimed to examine whether the P104L mutation affects C2C12 cell glucose metabolism, growth, and proliferation without insulin stimulation. C2C12 cells stably transfected with CAV3‐P104L were established, and biochemical assays, western blot analysis and confocal microscopy were used to observe glucose metabolism as well as cell growth and proliferation and to determine the effect of the P104L mutation on the PI3K/Akt signaling pathway. Without insulin stimulation, C2C12 cells stably transfected with the P104L CAV3 mutant exhibited decreased glucose uptake and glycogen synthesis, decreased CAV3 expression and reduced localization of CAV3 and GLUT4 on the cell membrane. The P104L mutant significantly reduced the cell diameters, but accelerated cell proliferation. Akt phosphorylation was inhibited, and protein expression of GLUT4, p‐GSK3β, and p‐p70s6K, which are molecules downstream of Akt, was significantly decreased. The CAV3‐P104L mutation inhibits glycometabolism and cell growth but accelerates C2C12 cell proliferation by reducing CAV3 protein expression and cell membrane localization, which may contribute to the pathogenesis of LGMD‐1C.

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“…Mice that lack caveolin-3 (cav3), a skeletal muscle-specific scaffolding protein critical in the formation of caveolae on the plasma membrane, exhibit skeletal muscle insulin resistance due to plasma membrane-specific effects on the IR (41, 42). Overexpression of dominant-negative cav3 leads to decreased glucose uptake and glycogen synthesis in C2C12 cells, which is attributed to decreased Akt phosphorylation (43–45). Conversely, an increase in wildtype cav3 expression is sufficient to enhance Akt phosphorylation and glucose uptake (46).…”

Section: Resultsmentioning

confidence: 99%

The Lands cycle modulates plasma membrane lipid organization and insulin sensitivity in skeletal muscle

Ferrara

Rong²,

Maschek

et al. 2019

Preprint

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39 Aberrant lipid metabolism promotes the development of skeletal muscle insulin resistance, but 40 the exact identity of lipid-mediated mechanisms relevant to human obesity remains unclear. A 41 comprehensive lipidomic analyses of primary myocytes from lean insulin-sensitive (LN) and 42 obese insulin-resistant (OB) individuals revealed several species of lysophospholipids (lyso-PL) 43 that were differentially-abundant. These changes coincided with greater expression of 44 lysophosphatidylcholine acyltransferase 3 (LPCAT3), an enzyme involved in phospholipid 45 transacylation (Lands cycle). Strikingly, mice with skeletal muscle-specific knockout of LPCAT3 46 (LPCAT3-MKO) exhibited greater muscle lyso-PC/PC, concomitant with greater insulin 47 sensitivity in vivo and insulin-stimulated skeletal muscle glucose uptake ex vivo. Absence of 48 LPCAT3 reduced phospholipid packing of the cellular membranes and increased plasma 49membrane lipid clustering, suggesting that LPCAT3 affects insulin receptor phosphorylation by 50 modulating plasma membrane lipid organization. In conclusion, obesity accelerates the skeletal 51 muscle Lands cycle, whose consequence might induce the disruption of plasma membrane 52 organization that suppresses muscle insulin action. 53Notably, the increase occurred at the level of the insulin receptor (IR), a node that is localized in 131 the phospholipid-rich plasma membrane. Consequently, LPCAT3 deletion enhanced insulin-132 stimulated glycogen synthesis ( Figure 2H), suggesting that this intervention increases skeletal 133 muscle insulin sensitivity in vitro (due to low GLUT4:GLUT1 stoichiometry, insulin-stimulated 134 glucose uptake is not an ideal surrogate for insulin sensitivity in C2C12 myotubes). LPCAT3 135 knockdown also enhanced insulin signaling in HSkMC from obese subjects ( Figure 2G). 136 137The organization and clustering of plasma membrane microdomains is linked to the induction of 138 tyrosine-kinase signaling events, such as IR signaling (29-31). Because LPCAT3 deletion 139

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The Caveolin-3 P104L mutation of LGMD-1C leads to disordered glucose metabolism in muscle cells

Cited by 14 publications

References 29 publications

Caveolin‐3 deficiency associated with the dystrophy P104L mutation impairs skeletal muscle mitochondrial form and function

Caveolin‐3 deficiency associated with the dystrophy P104L mutation impairs skeletal muscle mitochondrial form and function

The caveolin‐3 P104L mutation in LGMD‐1C patients inhibits non‐insulin‐stimulated glucose metabolism and growth but promotes myocyte proliferation

The Lands cycle modulates plasma membrane lipid organization and insulin sensitivity in skeletal muscle

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