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A thermolabile aldolase A mutant causes fever-induced recurrent rhabdomyolysis without hemolytic anemia.

Mamoune A, Bahuau M, Hamel Y, Serre V, Pelosi M, Habarou F, Nguyen Morel MA, Boisson B, Vergnaud S, Viou MT, Nonnenmacher L, Piraud M, Nusbaum P, Vamecq J, Romero N, Ottolenghi C, Casanova JL, de Lonlay P - PLoS Genet. (2014)

Bottom Line: Myoglobinuria was always triggered by febrile illnesses.Lipid droplets accumulated in patient myoblasts relative to control and this was increased by cytokines, and reduced by dexamethasone.We also propose a treatment for this severe disease.

View Article: PubMed Central - PubMed

Affiliation: INSERM U781, Institut Imagine des Maladies Génétiques, Université Paris Descartes et Centre de Référence des Maladies Héréditaires du Métabolisme, Hôpital Necker, AP-HP, Paris, France.

ABSTRACT
Aldolase A deficiency has been reported as a rare cause of hemolytic anemia occasionally associated with myopathy. We identified a deleterious homozygous mutation in the ALDOA gene in 3 siblings with episodic rhabdomyolysis without hemolytic anemia. Myoglobinuria was always triggered by febrile illnesses. We show that the underlying mechanism involves an exacerbation of aldolase A deficiency at high temperatures that affected myoblasts but not erythrocytes. The aldolase A deficiency was rescued by arginine supplementation in vitro but not by glycerol, betaine or benzylhydantoin, three other known chaperones, suggesting that arginine-mediated rescue operated by a mechanism other than protein chaperoning. Lipid droplets accumulated in patient myoblasts relative to control and this was increased by cytokines, and reduced by dexamethasone. Our results expand the clinical spectrum of aldolase A deficiency to isolated temperature-dependent rhabdomyolysis, and suggest that thermolability may be tissue specific. We also propose a treatment for this severe disease.

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Related in: MedlinePlus

ALDOA expression and activity.3A:ALDOA mRNA expression in control myoblasts (C, white bars) and the patient myoblasts (P, grey bars) under basal conditions, with TNFα+Ilβ treatment (left) or at a high temperature (right, 40°C); Aldolase A protein levels (lower panel) under basal conditions, with TNFα+Ilβ treatment or at a high temperature. 3B: Aldolase A activity in control and the patients' myoblasts under the same conditions: basal conditions, TNFα+Ilβ treatment and at different temperatures. The results are shown as the mean value ±SD from 3 independent experiments. 3C: Aldolase A activity in control and patients erythrocytes under basal conditions and at different temperatures. The results are shown as the mean value of two independent experiments. 3D: Aldolase A activity (upper) and protein level (below) in the patient myoblasts under basal condition and after arginine (Arg) treatment.*: p<0,05).
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pgen-1004711-g003: ALDOA expression and activity.3A:ALDOA mRNA expression in control myoblasts (C, white bars) and the patient myoblasts (P, grey bars) under basal conditions, with TNFα+Ilβ treatment (left) or at a high temperature (right, 40°C); Aldolase A protein levels (lower panel) under basal conditions, with TNFα+Ilβ treatment or at a high temperature. 3B: Aldolase A activity in control and the patients' myoblasts under the same conditions: basal conditions, TNFα+Ilβ treatment and at different temperatures. The results are shown as the mean value ±SD from 3 independent experiments. 3C: Aldolase A activity in control and patients erythrocytes under basal conditions and at different temperatures. The results are shown as the mean value of two independent experiments. 3D: Aldolase A activity (upper) and protein level (below) in the patient myoblasts under basal condition and after arginine (Arg) treatment.*: p<0,05).

Mentions: We studied whether temperature or pro-inflammatory cytokines affected ALDOA expression. To this end, the patient and the control myoblasts were cultured at 37 or 40°C, under basal conditions or with the combination of TNFα+IL-1β. Myoblasts from the patient and the controls responded to pro-inflammatory stress by significant secretion of IL6, peaking at approximately 24 hours of TNFα+IL-1ß stimulation (20-fold). The level of ALDOA mRNA was unchanged in patient myoblasts after exposure to TNFα+IL-1β or 40°C (Figure 3A upper panel), whereas the corresponding protein level was reduced in patient myoblasts in basal conditions (0.6±0.09; control: 1.3±0.17) and was further abated at 40°C (barely detectable) compared to control (0.8±0.16) (Figure 3A, lower panel). In contrast, TNFα+IL-1β treatment did not affect the protein level (1.2±0.11 to 0.97±0.11 in control and 0.7±0.09 to 0.8±0.13 in patient myoblasts). Accordingly, aldolase activity in the patient myoblasts dramatically decreased after incubation from 25°C, 37°C through 40°C (residual activity 10% and 5% respectively), and to a lesser degree in the control myoblasts (residual activity 61% at 37°C and 43% at 40°C) (Figure 3B). TNFα+IL-1β treatment did not change the level of activity in the patient or the control myoblasts (Figure 3B). Interestingly, aldolase A activity in erythrocytes from the three patients was not modified by high temperature (Figure 3C). These results suggest that the mutant enzyme might be differentially destabilized in distinct tissues, i.e., only in myoblasts and not in erythrocytes.


A thermolabile aldolase A mutant causes fever-induced recurrent rhabdomyolysis without hemolytic anemia.

Mamoune A, Bahuau M, Hamel Y, Serre V, Pelosi M, Habarou F, Nguyen Morel MA, Boisson B, Vergnaud S, Viou MT, Nonnenmacher L, Piraud M, Nusbaum P, Vamecq J, Romero N, Ottolenghi C, Casanova JL, de Lonlay P - PLoS Genet. (2014)

ALDOA expression and activity.3A:ALDOA mRNA expression in control myoblasts (C, white bars) and the patient myoblasts (P, grey bars) under basal conditions, with TNFα+Ilβ treatment (left) or at a high temperature (right, 40°C); Aldolase A protein levels (lower panel) under basal conditions, with TNFα+Ilβ treatment or at a high temperature. 3B: Aldolase A activity in control and the patients' myoblasts under the same conditions: basal conditions, TNFα+Ilβ treatment and at different temperatures. The results are shown as the mean value ±SD from 3 independent experiments. 3C: Aldolase A activity in control and patients erythrocytes under basal conditions and at different temperatures. The results are shown as the mean value of two independent experiments. 3D: Aldolase A activity (upper) and protein level (below) in the patient myoblasts under basal condition and after arginine (Arg) treatment.*: p<0,05).
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4230727&req=5

pgen-1004711-g003: ALDOA expression and activity.3A:ALDOA mRNA expression in control myoblasts (C, white bars) and the patient myoblasts (P, grey bars) under basal conditions, with TNFα+Ilβ treatment (left) or at a high temperature (right, 40°C); Aldolase A protein levels (lower panel) under basal conditions, with TNFα+Ilβ treatment or at a high temperature. 3B: Aldolase A activity in control and the patients' myoblasts under the same conditions: basal conditions, TNFα+Ilβ treatment and at different temperatures. The results are shown as the mean value ±SD from 3 independent experiments. 3C: Aldolase A activity in control and patients erythrocytes under basal conditions and at different temperatures. The results are shown as the mean value of two independent experiments. 3D: Aldolase A activity (upper) and protein level (below) in the patient myoblasts under basal condition and after arginine (Arg) treatment.*: p<0,05).
Mentions: We studied whether temperature or pro-inflammatory cytokines affected ALDOA expression. To this end, the patient and the control myoblasts were cultured at 37 or 40°C, under basal conditions or with the combination of TNFα+IL-1β. Myoblasts from the patient and the controls responded to pro-inflammatory stress by significant secretion of IL6, peaking at approximately 24 hours of TNFα+IL-1ß stimulation (20-fold). The level of ALDOA mRNA was unchanged in patient myoblasts after exposure to TNFα+IL-1β or 40°C (Figure 3A upper panel), whereas the corresponding protein level was reduced in patient myoblasts in basal conditions (0.6±0.09; control: 1.3±0.17) and was further abated at 40°C (barely detectable) compared to control (0.8±0.16) (Figure 3A, lower panel). In contrast, TNFα+IL-1β treatment did not affect the protein level (1.2±0.11 to 0.97±0.11 in control and 0.7±0.09 to 0.8±0.13 in patient myoblasts). Accordingly, aldolase activity in the patient myoblasts dramatically decreased after incubation from 25°C, 37°C through 40°C (residual activity 10% and 5% respectively), and to a lesser degree in the control myoblasts (residual activity 61% at 37°C and 43% at 40°C) (Figure 3B). TNFα+IL-1β treatment did not change the level of activity in the patient or the control myoblasts (Figure 3B). Interestingly, aldolase A activity in erythrocytes from the three patients was not modified by high temperature (Figure 3C). These results suggest that the mutant enzyme might be differentially destabilized in distinct tissues, i.e., only in myoblasts and not in erythrocytes.

Bottom Line: Myoglobinuria was always triggered by febrile illnesses.Lipid droplets accumulated in patient myoblasts relative to control and this was increased by cytokines, and reduced by dexamethasone.We also propose a treatment for this severe disease.

View Article: PubMed Central - PubMed

Affiliation: INSERM U781, Institut Imagine des Maladies Génétiques, Université Paris Descartes et Centre de Référence des Maladies Héréditaires du Métabolisme, Hôpital Necker, AP-HP, Paris, France.

ABSTRACT
Aldolase A deficiency has been reported as a rare cause of hemolytic anemia occasionally associated with myopathy. We identified a deleterious homozygous mutation in the ALDOA gene in 3 siblings with episodic rhabdomyolysis without hemolytic anemia. Myoglobinuria was always triggered by febrile illnesses. We show that the underlying mechanism involves an exacerbation of aldolase A deficiency at high temperatures that affected myoblasts but not erythrocytes. The aldolase A deficiency was rescued by arginine supplementation in vitro but not by glycerol, betaine or benzylhydantoin, three other known chaperones, suggesting that arginine-mediated rescue operated by a mechanism other than protein chaperoning. Lipid droplets accumulated in patient myoblasts relative to control and this was increased by cytokines, and reduced by dexamethasone. Our results expand the clinical spectrum of aldolase A deficiency to isolated temperature-dependent rhabdomyolysis, and suggest that thermolability may be tissue specific. We also propose a treatment for this severe disease.

Show MeSH
Related in: MedlinePlus