RESUMO
Short-chain acyl-CoA dehydrogenase (SCAD) is a mitochondrial enzyme involved in the beta-oxidation of fatty acids. Genetic defect of SCAD was documented to cause clinical symptoms such as progressive psychomotor retardation, muscle hypotonia, and myopathy in early reports. However, clinical significance of SCAD deficiency (SCADD) has been getting ambiguous, for some variants in the ACADS gene, which encodes the SCAD protein, has turned out to be widely prevailed among general populations. Accordingly, the pathophysiology of SCADD has not been clarified thus far. The present report focuses on two suspected cases of SCADD detected through the screening of newborns by tandem mass spectrometry. In both subjects, compound heterozygous mutations in ACADS were detected. The mutated genes were expressed in a transient gene expression system, and the enzymatic activities of the obtained mutant SCAD proteins were measured. The activities of the mutant SCAD proteins were significantly lower than that of the wild-type enzyme, confirming the mechanism underlying the diagnosis of SCADD in both subjects. Moreover, the mutant SCAD proteins gave rise to mitochondrial fragmentation and autophagy, both of which were proportional to the decrease in SCAD activities. The association of autophagy with programmed cell death suggests that the mutant SCAD proteins are toxic to mitochondria and to the cells in which they are expressed. The expression of recombinant ACADS-encoded mutant proteins offers a technique to evaluate both the nature of the defective SCAD proteins and their toxicity. Moreover, our results provide insight into possible molecular pathophysiology of SCADD.
Assuntos
Butiril-CoA Desidrogenase/deficiência , Butiril-CoA Desidrogenase/genética , Genes , Transtornos do Metabolismo dos Lipídeos/genética , Mutação , Erros Inatos do Metabolismo dos Aminoácidos/genética , Erros Inatos do Metabolismo dos Aminoácidos/metabolismo , Sequência de Bases , Butiril-CoA Desidrogenase/metabolismo , Genótipo , Heterozigoto , Humanos , Recém-Nascido , Transtornos do Metabolismo dos Lipídeos/metabolismo , Mitocôndrias/genética , Mitocôndrias/metabolismo , Dados de Sequência Molecular , Oxirredução , Estrutura Secundária de Proteína/genética , Proteínas Recombinantes/metabolismoRESUMO
Short-chain acylCoA dehydrogenase (SCAD) deficiency is a rare mitochondrial disorder involving the beta-oxidation of fatty acylCoA compounds in chains of 4-6 carbons. Unlike other mitochondrial disorders, cases involving autoimmune diseases have not been described. We report a 15-year-old boy with SCAD deficiency who suffered from pernicious anaemia, vitiligo, scleroatrophic lichen and autoimmune thyroiditis. As has been reported in other mitochondrial disorders, we hypothesised that autoimmune diseases are also present in SCAD deficiency. Furthermore, we discuss the possible pathogenetic relationship between these diseases.
Assuntos
Anemia Perniciosa/complicações , Doenças Autoimunes do Sistema Nervoso/genética , Butiril-CoA Desidrogenase/deficiência , Líquen Escleroso e Atrófico/complicações , Tireoidite Autoimune/complicações , Vitiligo/complicações , Adolescente , Anemia Perniciosa/genética , Humanos , Líquen Escleroso e Atrófico/genética , Masculino , Tireoidite Autoimune/genética , Vitiligo/genéticaRESUMO
Short-chain acyl-CoA dehydrogenase deficiency (SCADD) is a rare inherited disorder of the mitochondrial beta-oxidation of fatty acids. Patients with SCADD present mainly with symptoms of neuromuscular character. In order to investigate factors involved in the pathogenesis, we studied a disease-associated variant of the SCAD protein (p.Arg83Cys, c.319C>T), which is known to compromise SCAD protein folding. We investigated the consequences of overexpressing the misfolded mitochondrial protein, and thus determined whether the misfolded p.Arg83Cys SCAD proteins can elicit a toxic reaction. Human astrocytes were transiently transfected with either wild-type or p.Arg83Cys encoding cDNA, and analyzed for insoluble proteins/aggregate-formation, alterations in mitochondrial morphology, and for the presence of reactive oxygen species (ROS) in the mitochondria. The majority of cells overexpressing the p.Arg83Cys SCAD variant protein presented with an altered mitochondrial morphology of a grain-like structure, whereas the majority of the cells overexpressing wild-type SCAD presented with a normal thread-like mitochondrial reticulum. We found this grain-like structure to be associated with an increased amount of ROS. The mitochondrial morphology change was partly alleviated by addition of the mitochondrial targeted antioxidant MitoQ, indicating a ROS-induced mitochondrial fission. We therefore propose that SCAD misfolding leads to production of ROS, which in turn leads to fission and a grain-like structure of the mitochondrial reticulum. This finding indicates a toxic response elicited by misfolded p.Arg83Cys SCAD proteins.
Assuntos
Butiril-CoA Desidrogenase/química , Erros Inatos do Metabolismo Lipídico/genética , Mitocôndrias/genética , Estresse Oxidativo/genética , Dobramento de Proteína , Deficiências na Proteostase/genética , Astrócitos/enzimologia , Butiril-CoA Desidrogenase/deficiência , Butiril-CoA Desidrogenase/genética , Linhagem Celular , Humanos , Mitocôndrias/metabolismo , Mitocôndrias/ultraestrutura , Espécies Reativas de Oxigênio/metabolismo , TransfecçãoRESUMO
Short-chain acyl-CoA dehydrogenase deficiency (SCADD) is an inborn error, biochemically characterized by increased plasma butyrylcarnitine (C4-C) concentration and increased ethylmalonic acid (EMA) excretion and caused by rare mutations and/or common gene variants in the SCAD encoding gene. Although its clinical relevance is not clear, SCADD is included in most US newborn screening programs. Riboflavin, the precursor of flavin adenine dinucleotide (FAD, cofactor), might be effective for treating SCADD. We assessed the FAD status and evaluated the effects of riboflavin treatment in a prospective open-label cohort study involving 16 patients with SCADD, subdivided into mutation/mutation (mut/mut), mutation/variant (mut/var), and variant/variant (var/var) genotype groups. Blood FAD levels were normal in all patients before therapy, but significantly lower in the mut/var and var/var groups compared with the mut/mut group. Riboflavin treatment resulted in a decrease in EMA excretion in the mut/var group and in a subjective clinical improvement in four patients from this group. However, this improvement persisted after stopping treatment. These results indicate that high-dose riboflavin treatment may improve the biochemical features of SCADD, at least in patients with a mut/var genotype and low FAD levels. As our study could not demonstrate a clinically relevant effect of riboflavin, general use of riboflavin cannot be recommended.
Assuntos
Butiril-CoA Desidrogenase/deficiência , Flavina-Adenina Dinucleotídeo/sangue , Erros Inatos do Metabolismo Lipídico/tratamento farmacológico , Riboflavina/administração & dosagem , Complexo Vitamínico B/administração & dosagem , Adolescente , Biomarcadores/sangue , Biomarcadores/urina , Butiril-CoA Desidrogenase/genética , Carnitina/análogos & derivados , Carnitina/sangue , Criança , Pré-Escolar , Feminino , Flavina-Adenina Dinucleotídeo/urina , Predisposição Genética para Doença , Humanos , Lactente , Erros Inatos do Metabolismo Lipídico/sangue , Erros Inatos do Metabolismo Lipídico/enzimologia , Erros Inatos do Metabolismo Lipídico/genética , Erros Inatos do Metabolismo Lipídico/urina , Masculino , Malonatos/urina , Mutação , Fenótipo , Estudos Prospectivos , Resultado do TratamentoRESUMO
High concentrations of ethylmalonic acid are found in tissues and biological fluids of patients affected by ethylmalonic encephalopathy, deficiency of short-chain acyl-CoA dehydrogenase activity and other illnesses characterized by developmental delay and neuromuscular symptoms. The pathophysiological mechanisms responsible for the brain damage in these patients are virtually unknown. Therefore, in the present work we investigated the in vitro effect of EMA on oxidative stress parameters in rat cerebral cortex. EMA significantly increased chemiluminescence and thiobarbituric acid-reactive species levels (lipoperoxidation), as well as carbonyl content and oxidation of sulfhydryl groups (protein oxidative damage) and DCFH. EMA also significantly decreased the levels of reduced glutathione (non-enzymatic antioxidant defenses). In contrast, nitrate and nitrite levels were not affected by this short organic acid. It is therefore presumed that oxidative stress may represent a pathomechanism involved in the pathophysiology of the neurologic symptoms manifested by patients affected by disorders in which EMA accumulates.
Assuntos
Córtex Cerebral/efeitos dos fármacos , Malonatos/toxicidade , Estresse Oxidativo/efeitos dos fármacos , Animais , Antioxidantes/farmacologia , Butiril-CoA Desidrogenase/deficiência , Córtex Cerebral/metabolismo , Cromanos/farmacologia , Fluoresceínas/metabolismo , Glutationa/metabolismo , Peroxidação de Lipídeos/efeitos dos fármacos , Masculino , Oxirredução , Carbonilação Proteica/efeitos dos fármacos , Ratos , Ratos Wistar , Compostos de Sulfidrila/metabolismo , Substâncias Reativas com Ácido Tiobarbitúrico/metabolismoRESUMO
OBJECTIVE: To gain insight into the pathophysiological and clinical consequences of short-chain acyl-coenzyme A dehydrogenase deficiency (SCADD). STUDY DESIGN: A retrospective study of 15 fasting and 6 fat-loading tests in 15 Dutch patients with SCADD, divided into 3 genotype groups. Metabolic and endocrinologic measurements and the biochemical characteristics of SCADD, ethylmalonic acid (EMA), and C4-carnitine were studied. RESULTS: Three patients had development of hypoglycemia during fasting; all of these had originally presented with hypoglycemia. Metabolic and endocrinologic measurements remained normal during all tests. The EMA excretion increased in response to fasting and fat loading, and plasma C4-carnitine remained stable. Test results did not differ between the 3 genotype groups. CONCLUSIONS: The metabolic profiles of the 3 patients with development of hypoglycemia resemble idiopathic ketotic hypoglycemia. Because hypoglycemia generally requires a metabolic work-up and because SCADD is relatively prevalent, SCADD may well be diagnosed coincidently, thus being causally unrelated to the hypoglycemia. If SCADD has any other pathologic consequences, the accumulation of potentially toxic metabolites such as EMA is most likely involved. However, the results of our study indicate that there is no clear pathophysiological significance, irrespective of genotype, supporting the claim that SCADD is not suited for inclusion in newborn screening programs.
Assuntos
Butiril-CoA Desidrogenase/deficiência , Erros Inatos do Metabolismo/fisiopatologia , Adolescente , Criança , Pré-Escolar , Gorduras Insaturadas na Dieta/administração & dosagem , Jejum , Feminino , Humanos , Hipoglicemia/enzimologia , Lactente , Recém-Nascido , Masculino , Triagem Neonatal , Óleos de Plantas/administração & dosagem , Estudos Retrospectivos , Óleo de GirassolRESUMO
Mitochondrial fatty acid oxidation defects have been recognized since the early 1970s. The discovery rate has been rather constant, with 3-4 'new' disorders identified every decade and with the most recent example, ACAD9 deficiency, reported in 2007. In this presentation we will focus on three of the 'old' defects: medium-chain acyl-CoA dehydrogenase (MCAD) deficiency, riboflavin responsive multiple acyl-CoA dehydrogenation (RR-MAD) deficiency, and short-chain acyl-CoA dehydrogenase (SCAD) deficiency. These disorders have been discussed in many publications and at countless conference presentations, and many questions relating to them have been answered. However, continuing clinical and pathophysiological research has raised many further questions, and new ideas and methodologies may be required to answer these. We will discuss these challenges. For MCAD deficiency the key question is why 80% of symptomatic patients are homozygous for the prevalent ACADM gene variation c.985A > G whereas this is found in only approximately 50% of newborns with a positive screen. For RR-MAD deficiency, the challenge is to find the connection between variations in the ETFDH gene and the observed deficiency of a number of different mitochondrial dehydrogenases as well as deficiency of FAD and coenzyme Q(10). With SCAD deficiency, the challenge is to elucidate whether ACADS gene variations are disease-associated, especially when combined with other genetic/cellular/environmental factors, which may act synergistically.
Assuntos
Ácidos Graxos/metabolismo , Erros Inatos do Metabolismo/genética , Erros Inatos do Metabolismo/metabolismo , Doenças Mitocondriais/genética , Doenças Mitocondriais/metabolismo , Acil-CoA Desidrogenase/deficiência , Acil-CoA Desidrogenase/genética , Butiril-CoA Desidrogenase/deficiência , Butiril-CoA Desidrogenase/genética , Humanos , Modelos Biológicos , Deficiência Múltipla de Acil Coenzima A Desidrogenase/genética , Deficiência Múltipla de Acil Coenzima A Desidrogenase/metabolismo , Mutação de Sentido Incorreto , Oxirredução , Dobramento de ProteínaAssuntos
Butiril-CoA Desidrogenase/genética , Erros Inatos do Metabolismo Lipídico/enzimologia , Erros Inatos do Metabolismo Lipídico/genética , Triagem Neonatal , Avaliação de Resultados em Cuidados de Saúde , Butiril-CoA Desidrogenase/deficiência , Criança , Feminino , Testes Genéticos , Humanos , Recém-Nascido , Erros Inatos do Metabolismo Lipídico/diagnóstico , Masculino , MutaçãoRESUMO
An 8-month-old girl was admitted to an outpatient clinic with significant hypotonia and weakness. Organic acid analysis in urine revealed a significant increase in ethylmalonic acid. A deoxyribonucleic analysis revealed the presence of a 625G>A (G-to-A substitution at nucleotide 625) variant short-chain acyl-coenzyme A dehydrogenase gene polymorphism. With the clinical, biochemical and molecular findings, short-chain acyl-coenzyme A dehydrogenase deficiency was suspected. Because 625G>A and 511C>T (C-to-T substitution at nucleotide 511) genetic variations are also present in 14% of the general population, these are considered to be genetic sensitivity variations rather than causing a disease themselves and to result in possible short-chain acyl-coenzyme A dehydrogenase deficiency in the presence of environmental factors such as fever and hunger as well as cellular, biochemical, and other genetic factors. It was stressed that severe infantile hypotonia could also be the only manifestation of ethylmalonic aciduria spectrum disorders.
Assuntos
Alelos , Butiril-CoA Desidrogenase/deficiência , Butiril-CoA Desidrogenase/genética , Análise Mutacional de DNA , Erros Inatos do Metabolismo Lipídico/genética , Malonatos/urina , Hipotonia Muscular/genética , Debilidade Muscular/genética , Deficiências do Desenvolvimento/diagnóstico , Deficiências do Desenvolvimento/genética , Deficiências do Desenvolvimento/urina , Diagnóstico Diferencial , Feminino , Genótipo , Humanos , Lactente , Erros Inatos do Metabolismo Lipídico/diagnóstico , Erros Inatos do Metabolismo Lipídico/urina , Hipotonia Muscular/diagnóstico , Hipotonia Muscular/urina , Debilidade Muscular/diagnóstico , Debilidade Muscular/urina , Exame Neurológico , Fenótipo , Polimorfismo Genético/genéticaRESUMO
Recombinant adeno-associated viral vectors pseudotyped with serotype 5 and 8 capsids (AAV5 and AAV8) have been shown to be efficient gene transfer reagents for the liver. We have produced AAV5 and AAV8 vectors that express mouse short-chain acyl-CoA dehydrogenase (mSCAD) cDNA under the transcriptional control of the cytomegalovirus-chicken beta-actin hybrid promoter. We hypothesized that these vectors would produce sufficient hepatocyte transduction (after administration via the portal vein) and thus sufficient SCAD enzyme to correct the phenotype observed in the SCAD-deficient (BALB/cByJ) mouse, which includes elevated blood butyrylcarnitine and hepatic steatosis. Ten weeks after portal vein injection into 8-week-old mice, AAV8-treated livers contained acyl-CoA dehydrogenase activity (14.3 mU/mg) toward butyryl-CoA, compared with 7.6 mU/mg in mice that received phosphate-buffered saline. Immunohistochemistry showed expression of mSCAD within rAAV8-mSCAD-transduced hepatocytes, as seen by light microscopy. A significant reduction of circulating butyrylcarnitine was seen in AAV5-mSCAD- and AAV8-mSCAD-injected mice. Magnetic resonance spectroscopy of fasted mice demonstrated a significant reduction in relative lipid content within the livers of AAV8-mSCAD-treated mice. These results demonstrate biochemical correction of SCAD deficiency after AAV8-mediated SCAD gene delivery.
Assuntos
Butiril-CoA Desidrogenase/deficiência , Butiril-CoA Desidrogenase/genética , Dependovirus , Terapia Genética , Fígado , Erros Inatos do Metabolismo/terapia , Actinas/genética , Animais , Carnitina/análogos & derivados , Carnitina/sangue , Citomegalovirus/genética , Vetores Genéticos , Injeções Intravenosas , Fígado/irrigação sanguínea , Camundongos , Camundongos Mutantes , Veia Porta , Regiões Promotoras Genéticas , Transdução Genética/métodosRESUMO
The aim of the current study was to assess lipid metabolism in horses with atypical myopathy. Urine samples from 10 cases were subjected to analysis of organic acids, glycine conjugates, and acylcarnitines revealing increased mean excretion of lactic acid, ethylmalonic acid, 2-methylsuccinic acid, butyrylglycine, (iso)valerylglycine, hexanoylglycine, free carnitine, C2-, C3-, C4-, C5-, C6-, C8-, C8:1-, C10:1-, and C10:2-carnitine as compared with 15 control horses (12 healthy and three with acute myopathy due to other causes). Analysis of plasma revealed similar results for these predominantly short-chain acylcarnitines. Furthermore, measurement of dehydrogenase activities in lateral vastus muscle from one horse with atypical myopathy indeed showed deficiencies of short-chain acyl-CoA dehydrogenase (0.66 as compared with 2.27 and 2.48 in two controls), medium-chain acyl-CoA dehydrogenase (0.36 as compared with 4.31 and 4.82 in two controls) and isovaleryl-CoA dehydrogenase (0.74 as compared with 1.43 and 1.61 nmol min(-1) mg(-1) in two controls). A deficiency of several mitochondrial dehydrogenases that utilize flavin adenine dinucleotide as cofactor including the acyl-CoA dehydrogenases of fatty acid beta-oxidation, and enzymes that degrade the CoA-esters of glutaric acid, isovaleric acid, 2-methylbutyric acid, isobutyric acid, and sarcosine was suspected in 10 out of 10 cases as the possible etiology for a highly fatal and prevalent toxic equine muscle disease similar to the combined metabolic derangements seen in human multiple acyl-CoA dehydrogenase deficiency also known as glutaric acidemia type II.
Assuntos
Acil-CoA Desidrogenases/deficiência , Doenças dos Cavalos/metabolismo , Doenças Musculares/metabolismo , Acil-CoA Desidrogenase/deficiência , Acil-CoA Desidrogenase/metabolismo , Acil-CoA Desidrogenases/metabolismo , Animais , Ácido Butírico/sangue , Ácido Butírico/urina , Butiril-CoA Desidrogenase/deficiência , Butiril-CoA Desidrogenase/metabolismo , Carnitina/análogos & derivados , Carnitina/sangue , Carnitina/urina , Cromatografia Líquida de Alta Pressão , Feminino , Cromatografia Gasosa-Espectrometria de Massas , Glutaratos/sangue , Glutaratos/urina , Doenças dos Cavalos/enzimologia , Doenças dos Cavalos/patologia , Cavalos , Isovaleril-CoA Desidrogenase/deficiência , Isovaleril-CoA Desidrogenase/metabolismo , Ácido Láctico/sangue , Ácido Láctico/urina , Masculino , Microscopia Eletrônica , Microscopia de Fluorescência , Músculos/patologia , Músculos/ultraestrutura , Doenças Musculares/enzimologia , Doenças Musculares/patologia , Riboflavina/sangueRESUMO
2-Methylbutyryl-CoA dehydrogenase (MBD; coded by the ACADSB gene) catalyzes the step in isoleucine metabolism that corresponds to the isovaleryl-CoA dehydrogenase reaction in the degradation of leucine. Deficiencies of both enzymes may be detected by expanded neonatal screening with tandem-mass spectrometry due to elevated pentanoylcarnitine (C5 acylcarnitine) in blood, but little information is available on the clinical relevance of MBD deficiency. We biochemically and genetically characterize six individuals with MBD deficiency from four families of different ethnic backgrounds. None of the six individuals showed clinical symptoms attributable to MBD deficiency although the defect in isoleucine catabolism was demonstrated both in vivo and in vitro. Several mutations in the ACADSB gene were identified, including a novel one. MBD deficiency may be a harmless metabolic variant although significant impairment of valproic acid metabolism cannot be excluded and further study is required to assess the long-term outcome of individuals with this condition. The relatively high prevalence of ACADSB gene mutations in control subjects suggests that MBD deficiency may be more common than previously thought but is not detected because of its usually benign nature.
Assuntos
Erros Inatos do Metabolismo dos Aminoácidos/diagnóstico , Erros Inatos do Metabolismo dos Aminoácidos/genética , Butiril-CoA Desidrogenase/deficiência , Isoleucina/metabolismo , Erros Inatos do Metabolismo dos Aminoácidos/metabolismo , Erros Inatos do Metabolismo dos Aminoácidos/urina , Butiril-CoA Desidrogenase/genética , Células Cultivadas , Criança , Pré-Escolar , Feminino , Genótipo , Glicina/análogos & derivados , Glicina/urina , Humanos , Lactente , Isoleucina/administração & dosagem , Masculino , Metabolismo , Valeratos/urinaRESUMO
We report 10 children (7 male, 3 female), 3 homozygous for c.319C>T mutation and 7 heterozygous for c.319C>T on one allele and c.625G>A variant on the other in the short-chain acyl-CoA dehydrogenase (SCAD) gene (ACADS). All were of Ashkenazi Jewish origin in which group we found a c.319C>T heterozygote frequency of 1:15 suggesting the presence of a founder mutation or selective advantage. Phenotype was variable with onset from birth to early childhood. Features included hypotonia (8/10), developmental delay (8/10), myopathy (4/10) with multicore changes in two and lipid storage in one, facial weakness (3/10), lethargy (5/10), feeding difficulties (4/10) and congenital abnormalities (3/7). One female with multiminicore myopathy had progressive external ophthalmoplegia, ptosis and cardiomyopathy with pneumonia and respiratory failure. Two brothers presented with psychosis, pyramidal signs, and multifocal white matter abnormalities on MRI brain suggesting additional genetic factors. Two other infants also had white matter changes. Elevated butyrylcarnitine (4/8), ethylmalonic aciduria (9/9), methylsuccinic aciduria (6/7), decreased butyrate oxidation in lymphoblasts (2/4) and decreased SCAD activity in fibroblasts or muscle (3/3) were shown. Expression studies of c.319C>T in mouse liver mitochondria showed it to be inactivating. c.625G>A is a common variant in ACADS that may confer disease susceptibility. Five healthy parents were heterozygous for c.319C>T and c.625G>A, suggesting reduced penetrance or broad clinical spectrum. We conclude that the c.319C>T mutation can lead to wide clinical and biochemical phenotypic variability, suggesting a complex multifactorial/polygenic condition. This should be screened for in individuals with multicore myopathy, particularly among the Ashkenazim.
Assuntos
Butiril-CoA Desidrogenase/deficiência , Butiril-CoA Desidrogenase/genética , Efeito Fundador , Judeus/genética , Mutação Puntual , Anormalidades Múltiplas/enzimologia , Anormalidades Múltiplas/genética , Adolescente , Adulto , Alelos , Animais , Sequência de Bases , Criança , Pré-Escolar , Primers do DNA/genética , Feminino , Heterozigoto , Homozigoto , Humanos , Lactente , Recém-Nascido , Masculino , Erros Inatos do Metabolismo/enzimologia , Erros Inatos do Metabolismo/genética , Camundongos , Doenças Musculares/enzimologia , Doenças Musculares/genética , Fenótipo , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismoRESUMO
This report presents a case of short-chain acyl-coenzyme A (CoA) dehydrogenase deficiency with a previously unreported presentation with brain malformations and infantile spasms. This female infant developed repeated tonic clonic seizures at the age of 3(1/2) months. She subsequently developed West syndrome at the age of 4 months. Her electroencephalogram disclosed hypsarrhythmia, and video-electroencephalographic monitoring confirmed the presence of infantile spasms. Magnetic resonance imaging revealed a small midline frontal meningocele, abnormal cortical gyration, and partial agenesis of the corpus callosum consistent with neuronal migrational disorder. Metabolic evaluation indicated ethylmalonic acidemia. Muscle biopsy with enzymatic assay of short-chain acyl-coenzyme A revealed low enzymatic activity confirming the diagnosis of short-chain acyl-coenzyme A dehydrogenase deficiency. To our knowledge, this is the first report of the coexistence of short-chain acyl-coenzyme A dehydrogenase deficiency, infantile spasms, and brain malformation. We conclude that short-chain acyl-coenzyme A dehydrogenase deficiency should be considered in the differential diagnosis of gyral abnormality, corpus callosal hypoplasia, and infantile spasms.
Assuntos
Agenesia do Corpo Caloso , Butiril-CoA Desidrogenase/deficiência , Erros Inatos do Metabolismo/complicações , Espasmos Infantis/etiologia , Espasmos Infantis/patologia , Córtex Cerebral/anormalidades , Pré-Escolar , Deficiências do Desenvolvimento/etiologia , Deficiências do Desenvolvimento/patologia , Epilepsia Tônico-Clônica/etiologia , Epilepsia Tônico-Clônica/patologia , Feminino , Humanos , Lactente , Imageamento por Ressonância MagnéticaRESUMO
Short-chain-acyl-CoA-dehydrogenase (SCAD) deficiency is an inborn error of mitochondrial fatty acid metabolism caused by rare mutations as well as common susceptibility variations in the SCAD gene. We describe the case of a 23-year-old male patient who had growth and mental retardation, recurrent vomiting, fever and seizures since infancy. Urinary gas chromatography and (1)H-nuclear magnetic resonance showed elevated levels of ethylmalonic acid. Serum concentrations of acylcarnitine, especially butyrylcarnitine (C4), were abnormally high. A homozygous variant allele of the SCAD gene, 625G>A, was detected. The patient broadens the clinical phenotype of SCAD deficiency and underlines the difficulty of diagnosis. The limited number of patients described may be the result of underdiagnosis.
Assuntos
Butiril-CoA Desidrogenase/deficiência , Espectroscopia de Ressonância Magnética/métodos , Erros Inatos do Metabolismo/sangue , Adulto , Butiril-CoA Desidrogenase/genética , Carnitina/análogos & derivados , Carnitina/sangue , Cromatografia Gasosa/métodos , Humanos , Masculino , Erros Inatos do Metabolismo/diagnóstico , Erros Inatos do Metabolismo/genéticaRESUMO
High concentrations of ethylmalonic acid (EMA) occur in tissues and biological fluids of patients affected by deficiency of short-chain acyl-CoA dehydrogenase activity, as well as in other illnesses characterized by neurological and muscular symptoms. Considering that the pathophysiological mechanisms responsible for the clinical manifestations of these diseases are virtually unknown, in the present work we developed a chemical in vivo model of ethylmalonic acidemia in young Wistar rats for neurochemical and behavioral studies through subcutaneous administration of EMA to young rats. The doses of EMA administered subcutaneously varied according to the age of the animals, being injected 3, 4, and 6 micromol g(-1) of body weight in rats of 7, 14, and 21 days, respectively. The concentrations of the acid were measured in blood and brain at regular intervals after a single injection (30-120 min) and reached the highest concentrations (3.0 mM and 0.5 micromol g(-1), approximately 0.5 mM), respectively, after 30 and 60 min of EMA injection. Next, we investigated the effects of acute EMA administration on the activities of complexes I-III, II, II-III, and IV of the respiratory chain in cerebral cortex and skeletal muscle, as well as on the activity of creatine kinase in cerebral cortex, striatum, skeletal muscle, and cardiac muscle of rats of 14 days of life. Control rats were treated with saline in the same volumes. We verified EMA administration did not change these enzymatic activities in all tissues studied. Although transient high concentrations of EMA did not alter important parameters of energy metabolism, it cannot be ruled out that chronic administration of this organic acid would disrupt energy metabolism.
Assuntos
Córtex Cerebral/efeitos dos fármacos , Córtex Cerebral/enzimologia , Metabolismo Energético/efeitos dos fármacos , Malonatos/farmacocinética , Acidose/metabolismo , Fatores Etários , Animais , Butiril-CoA Desidrogenase/deficiência , Complexo I de Transporte de Elétrons/metabolismo , Complexo II de Transporte de Elétrons/metabolismo , Complexo III da Cadeia de Transporte de Elétrons/metabolismo , Complexo IV da Cadeia de Transporte de Elétrons/metabolismo , Metabolismo Energético/fisiologia , Ativação Enzimática/efeitos dos fármacos , Ativação Enzimática/fisiologia , Feminino , Injeções Subcutâneas , Masculino , Malonatos/sangue , Músculo Esquelético/efeitos dos fármacos , Músculo Esquelético/enzimologia , Miocárdio/enzimologia , Gravidez , Ratos , Ratos WistarRESUMO
BACKGROUND: Hyperglycemic hyperosmolar nonketotic syndrome (HHNS) is usually associated with type 2 diabetes mellitus and is rare in children. However, a fatal malignant hyperthermia-like syndrome (MHLS) with rhabdomyolysis associated with new-onset diabetes mellitus and HHNS in adolescents has been described. DESIGN/METHODS: Case series. RESULTS: A 16-yr-old obese male (case A) and a 10-yr-old mid-pubertal nonobese female (case B) presented within a 6-month period with emesis, altered mental status, blood glucose >1600 mg/dL, and laboratory evidence of rhabdomyolysis. Case A developed fever after initiation of insulin therapy, along with refractory hypotension and multiorgan failure. He died 14 hrs after admission. Case B developed fever before insulin therapy, was treated with dantrolene, and made a full recovery. Metabolic workup showed evidence of short-chain acyl-CoA dehydrogenase (SCAD) deficiency. CONCLUSIONS: We report two cases of malignant hyperthermia-like syndrome associated with HHNS in adolescents. Their respective fluid management and clinical courses are described. Dantrolene therapy should be initiated immediately after this syndrome is recognized. We believe it is unlikely insulin is the sole trigger for MHLS. Case B is unique in that there was evidence of SCAD deficiency, a metabolic defect that we propose could lead to MHLS. We recommend that all patients with HHNS and MHLS be evaluated for an underlying metabolic disorder.
Assuntos
Diabetes Mellitus Tipo 2/complicações , Coma Hiperglicêmico Hiperosmolar não Cetótico/terapia , Hipertermia Maligna/etiologia , Hipertermia Maligna/terapia , Adolescente , Negro ou Afro-Americano , Butiril-CoA Desidrogenase/deficiência , Criança , Dantroleno/uso terapêutico , Evolução Fatal , Feminino , Hidratação , Humanos , Coma Hiperglicêmico Hiperosmolar não Cetótico/etiologia , Masculino , Obesidade/complicações , Rabdomiólise/etiologia , Síndrome , Resultado do TratamentoRESUMO
Mitochondrial beta-oxidation of fatty acids is required to meet physiologic energy requirements during illness and periods of fasting or physiologic stress, and is most active in liver and striated muscle. Acyl-CoA dehydrogenases of varying chain-length specificities represent the first step in the mitochondria for each round of beta-oxidation, each of which removes two-carbon units as acetyl-CoA for entry into the tricarboxylic acid cycle. We have used recombinant adeno-associated virus (rAAV) vectors expressing short-chain acyl-CoA dehydrogenase (SCAD) to correct the accumulation of fatty acyl-CoA intermediates in deficient cell lines. The rAAV-SCAD vector was then packaged into either rAAV serotype 1 or 2 capsids and injected intramuscularly into SCAD-deficient mice. A systemic effect was observed as judged by restoration of circulating butyryl- carnitine levels to normal. Total lipid content at the injection site was also decreased as demonstrated by noninvasive magnetic resonance spectroscopy (MRS). SCAD enzyme activity in the injected muscle was found at necropsy to be above the normal control mouse level. This study is the first to demonstrate the systemic correction of a fatty acid oxidation disorder with rAAV and the utility of MRS as a noninvasive method to monitor SCAD correction after in vivo gene therapy.
Assuntos
Dependovirus/fisiologia , Ácidos Graxos/metabolismo , Terapia Genética/métodos , Vetores Genéticos/fisiologia , Erros Inatos do Metabolismo Lipídico/terapia , Animais , Butiril-CoA Desidrogenase/deficiência , Butiril-CoA Desidrogenase/genética , Butiril-CoA Desidrogenase/metabolismo , Carnitina/análogos & derivados , Carnitina/análise , Carnitina/sangue , Linhagem Celular , DNA Recombinante , Dependovirus/enzimologia , Ácidos Graxos/análise , Feminino , Fibroblastos/metabolismo , Humanos , Injeções Intramusculares , Espectrometria de Massas/métodos , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Mitocôndrias/metabolismo , Músculos/química , Músculos/enzimologia , Oxirredução , Reprodutibilidade dos Testes , Transdução GenéticaRESUMO
Short/branched-chain acyl-CoA dehydrogenase deficiency (SBCADD) is an autosomal recessive disorder of L: -isoleucine catabolism. Little is known about the clinical presentation associated with this enzyme defect, as it has been reported in only a limited number of patients. Because the presence of C5-carnitine in blood may indicate SBCADD, the disorder may be detected by MS/MS-based routine newborn screening. It is, therefore, important to gain more knowledge about the clinical presentation and the mutational spectrum of SBCADD. In the present study, we have studied two unrelated families with SBCADD, both with seizures and psychomotor delay as the main clinical features. One family illustrates the fact that affected individuals may also remain asymptomatic. In addition, the normal level of newborn blood spot C5-acylcarnitine in one patient underscores the fact that newborn screening by MS/MS currently lacks sensitivity in detecting SBCADD. Until now, seven mutations in the SBCAD gene have been reported, but only three have been tested experimentally. Here, we identify and characterize an IVS3+3A>G mutation (c.303+3A>G) in the SBCAD gene, and provide evidence that this mutation is disease-causing in both families. Using a minigene approach, we show that the IVS3+3A>G mutation causes exon 3 skipping, despite the fact that it does not appear to disrupt the consensus sequence of the 5' splice site. Based on these results and numerous literature examples, we suggest that this type of mutation (IVS+3A>G) induces missplicing only when in the context of non-consensus (weak) 5' splice sites. Statistical analysis of the sequences shows that the wild-type versions of 5' splice sites in which +3A>G mutations cause exon skipping and disease are weaker on average than a random set of 5' splice sites. This finding is relevant to the interpretation of the functional consequences of this type of mutation in other disease genes.
Assuntos
Processamento Alternativo/genética , Butiril-CoA Desidrogenase/deficiência , Butiril-CoA Desidrogenase/genética , Isoleucina/metabolismo , Mutação , Células Cultivadas , Análise Mutacional de DNA , HumanosRESUMO
We have used mice with inborn errors of mitochondrial fatty acid beta-oxidation to test the concept of synergistic heterozygosity. We postulated that clinical disease can result from heterozygous mutations in more than one gene in single or related metabolic pathways. Mice with combinations of mutations in mitochondrial fatty acid beta-oxidation genes were cold challenged to test their ability to maintain normal body temperature, a sensitive indicator of overall beta-oxidation function. This included mice of the following genotypes: triple heterozygosity for mutations in very-long-chain acyl CoA dehydrogenase, long-chain acyl CoA dehydrogenase, and short-chain acyl CoA dehydrogenase genes (VLCAD+/-//LCAD+/-//SCAD+/-); double heterozygosity for mutations in VLCAD and LCAD genes (VLCAD+/-//LCAD+/-); double heterozygosity for mutations in LCAD and SCAD genes (LCAD+/-//SCAD+/-); single heterozygous mice (VLCAD+/-, LCAD+/-, SCAD+/-) and wild-type. We found that approximately 33% of mice with any of the combined mutant genotypes tested became hypothermic during a cold challenge. All wild-type and single heterozygous mice maintained normal body temperature throughout a cold challenge. Despite development of hypothermia in some double heterozygous mice, blood glucose concentrations remained normal. Biochemical screening by acylcarnitine and fatty acid analyses demonstrated results that varied by genotype. Thus, physiologic reduction of the beta-oxidation pathway, characterized as cold intolerance, occurred in mice with double or triple heterozygosity; however, the derangement was milder than in mice homozygous for any of these mutations. These results substantiate the concept of synergistic heterozygosity and illustrate the potential complexity involved in diagnosis and characterization of inborn errors of fatty acid metabolism in humans.