RESUMEN
Several lines of evidence implicate dysfunction in brain energy production as a key component of bipolar disorder. In particular, elevated brain lactate levels observed in this condition suggest a shift from aerobic to anaerobic metabolism, possibly as a result of mitochondrial abnormalities. Most prior imaging studies of brain metabolites were performed in either euthymic or depressed bipolar patients or compared different populations in different mood states. We sought to measure brain metabolite concentrations in the same patients in both manic and euthymic states. Given the dramatic changes in clinical state of bipolar disorder patients, we hypothesized that previously observed abnormalities in lactate concentrations in bipolar disorder might show state dependent changes. In this study 15 patients (mean age 36.1 years) diagnosed with bipolar I disorder underwent proton magnetic resonance spectroscopy of the anterior cingulate cortex and parieto-occipital cortex during hospitalization for acute mania (mean Young Mania Rating Scale (YMRS) 22.1). Seven of these subjects returned (mean interval 21.16 months) to have imaging repeated while euthymic (mean YMRS 2.0). A group of age- and gender-matched control participants (N=6) were scanned as well. We report that during mania, bipolar disorder subjects had lactate levels comparable to healthy control subjects but during euthymia these levels were significantly reduced. No significant change was observed for other metabolites. These results implicate mood dependent alterations in energy metabolism in the biology of bipolar disorder. Additionally, this finding has potential use as a biomarker for both evaluating novel treatments as well as diagnostic clarification between mood disorders.
Asunto(s)
Trastorno Bipolar/patología , Encéfalo/patología , Espectroscopía de Resonancia Magnética/métodos , Adulto , Afecto , Trastorno Bipolar/metabolismo , Trastorno Bipolar/psicología , Encéfalo/metabolismo , Metabolismo Energético , Femenino , Giro del Cíngulo/metabolismo , Giro del Cíngulo/patología , Humanos , Procesamiento de Imagen Asistido por Computador/métodos , Ácido Láctico/metabolismo , Estudios Longitudinales , Masculino , Proyectos Piloto , Escalas de Valoración PsiquiátricaRESUMEN
Fabry disease (OMIM 301500) is a rare X-linked recessive disorder caused by mutations in the alpha-galactosidase gene (GLA). Loss of alpha-galactosidase (alpha-Gal) activity leads to the abnormal accumulation of glycosphingolipids in lysosomes predominantly of vascular endothelial cells. Clinically the disorder presents with angiokeratomas, clouding of the cornea, and renal, cardiac, and cerebrovascular complications. In addition, there is an increased incidence of neuropathic pain in Fabry patients. In this study, we investigated the implications of loss of alpha-galactosidase A activity on sensorimotor function and peripheral nervous system. Similar to the described in Fabry disease patients, the sensorimotor assessment of Fabry mice revealed diminished locomotor activity and warm hypoalgesia as assessed in the hot-plate. Moreover Fabry mice displayed alterations both in balance and co-ordination. By histological analysis, the cyto-architecture of Fabry mice sciatic nerves showed an increase in mean cross-sectional area accompanied by a decrease in the density of non-myelinated fibers as well as a trend for a decreased number of small myelinated fibers, a well established feature of Fabry disease. A relative preservation of large myelinated fibers and nerve conduction velocity measurements was observed. Our findings demonstrate for the first time that Fabry knockout mice have Gb3 accumulation in the peripheral nervous system, alterations in sensorimotor function, hypoalgesia and no impairment of motor nerve conduction.
Asunto(s)
Enfermedad de Fabry , Sistema Nervioso Periférico/patología , Sistema Nervioso Periférico/fisiopatología , alfa-Galactosidasa/genética , Animales , Ataxia/fisiopatología , Modelos Animales de Enfermedad , Enfermedad de Fabry/patología , Enfermedad de Fabry/fisiopatología , Enfermedad de Fabry/psicología , Femenino , Glicoesfingolípidos/metabolismo , Calor , Hipoestesia/fisiopatología , Masculino , Ratones , Ratones Noqueados , Actividad Motora , Fibras Nerviosas/patología , Fibras Nerviosas/fisiología , Fibras Nerviosas/ultraestructura , Conducción Nerviosa/fisiología , Dolor/fisiopatología , Sistema Nervioso Periférico/ultraestructura , Fenotipo , Equilibrio Postural , Nervio Ciático/patología , Nervio Ciático/fisiopatología , Nervio Ciático/ultraestructura , Sensación Térmica , Trihexosilceramidas/metabolismo , alfa-Galactosidasa/metabolismoRESUMEN
A deficiency in alpha-galactosidase A (alpha-gal A) activity causes Fabry disease. Virus-based delivery of genes can correct cells and establish a sustained supply of therapeutic proteins. Recombinant lentiviral vectors (LVs) show promise in this context. We first demonstrate LV-mediated marking of peripheral blood (PB) cells by transduction/transplantation of hematopoietic stem/progenitor cells. Stable enGFP expression was observed in PB for 37 weeks. Next, we transplanted Fabry mice with bone marrow mononuclear cells (BMMNCs) transduced a single time with a LV encoding the human alpha-gal A cDNA. Sustained expression of functional alpha-gal A in Fabry mice was observed over 24 weeks. Plasma alpha-gal A activity from treated Fabry mice was two-fold higher than wild-type controls. Increased alpha-gal A activity, often to supra-normal levels, and reduction of globotriaosylceramide, a glycolipid that accumulates in Fabry disease, was observed in all organs assessed. In secondary bone marrow transplantations, Fabry mice showed multilineage marking of PB, splenocytes and BMMNCs, along with therapeutic levels of alpha-gal A activity in plasma and organs over 20 weeks. Lastly, we transduced mobilized PB CD34(+) cells from a Fabry patient and observed corresponding enzymatic increases. Thus a single LV-mediated transduction of primitive hematopoietic cells can result in sustained correction for Fabry disease.
Asunto(s)
Enfermedad de Fabry/terapia , Terapia Genética/métodos , Células Madre Hematopoyéticas/enzimología , alfa-Galactosidasa/genética , Animales , Linfocitos B/enzimología , Enfermedad de Fabry/enzimología , Citometría de Flujo , Vectores Genéticos/administración & dosificación , Vectores Genéticos/genética , Trasplante de Células Madre Hematopoyéticas , Humanos , Lentivirus/genética , Ratones , Ratones Noqueados , Modelos Animales , Transducción Genética/métodos , alfa-Galactosidasa/metabolismoRESUMEN
OBJECTIVE: To determine if convection-enhanced delivery (CED) of glucocerebrosidase could be used to treat targeted sites of disease progression in the brain and brainstem of a patient with neuronopathic Gaucher disease while monitoring enzyme distribution using MRI. METHODS: A CED paradigm in rodents (n = 8) and primates (n = 5) that employs co-infusion of a surrogate MRI tracer (gadolinium diethylenetriamine penta-acetic acid [Gd-DTPA]) with glucocerebrosidase to permit real-time monitoring of distribution was developed. The safety and feasibility of this delivery and monitoring paradigm were evaluated in a patient with type 2 Gaucher disease. RESULTS: Animal studies revealed that real-time, T1-weighted, MRI of Gd-DTPA accurately tracked enzyme distribution during CED. Targeted perfusion of clinically affected anatomic sites in a patient with neuronopathic Gaucher disease (frontal lobe and brainstem) with glucocerebrosidase was successfully performed. Real-time MRI revealed progressive and complete filling of the targeted region with enzyme and Gd-DTPA infusate. The patient tolerated the infusions without evidence of toxicity. CONCLUSIONS: Convection-enhanced delivery can be used to safely perfuse large regions of the brain and brainstem with therapeutic levels of glucocerebrosidase. Co-infused imaging surrogate tracers can be used to monitor and control the distribution of therapeutic agents in vivo. Patients with neuronopathic Gaucher disease and other intrinsic CNS disorders may benefit from a similar treatment paradigm.
Asunto(s)
Convección , Enfermedad de Gaucher/tratamiento farmacológico , Enfermedad de Gaucher/cirugía , Glucosilceramidasa/administración & dosificación , Cirugía Asistida por Computador/métodos , Animales , Enfermedad de Gaucher/diagnóstico por imagen , Humanos , Lactante , Macaca mulatta , Masculino , Neuronas/efectos de los fármacos , Neuronas/patología , Radiografía , Ratas , Ratas Sprague-DawleyRESUMEN
Fabry disease, OMIM 301500, is a progressive multisystem storage disorder due to the deficiency of alpha-galactosidase A (GALA). Neurological and vascular manifestations of this disorder with regard to hearing loss have not been analysed quantitatively in large cohorts. We conducted a retrospective cross sectional analysis of hearing loss in 109 male and female patients with Fabry disease who were referred to and seen at the Clinical Center of the National Institutes of Health, Bethesda, MD, USA on natural history and enzyme replacement study protocols. There were 85 males aged 6-58 years (mean 31 years, SD 13) and 24 females aged 22-72 years (mean 42 years, SD 12). All patients underwent a comprehensive audiological evaluation. In addition, cerebral white matter lesions, peripheral neuropathy, and kidney function were quantitatively assessed. HL(95), defined as a hearing threshold above the 95th percentile for age and gender matched normal controls, was present in 56% [95% CI (42.2-67.2)] of the males. Prevalence of HL(95) was lower in the group of patients with residual GALA enzyme activity compared with those without detectable activity (33% versus 63%) HL(95) was present in the low-, mid- and high-frequency ranges for all ages. Male patients with HL(95) had a higher microvascular cerebral white matter lesion load [1.4, interquartile range (IQR) 0-30.1 +/- versus 0, IQR 0-0], more pronounced cold perception deficit [19.4 +/- 5.5 versus 13.5 +/- 5.5 of just noticeable difference (JND) units] and lower kidney function [creatinine: 1.6 +/- 1.2 versus 0.77 +/- 0.2 mg/dl; blood urea nitrogen (BUN): 20.1 +/- 14.1 versus 10.3 +/- 3.28 mg/dl] than those without HL(95) (P < 0.001). Of the females, 38% had HL(95). There was no significant association with cold perception deficit, creatinine or BUN in the females. Word recognition and acoustic reflexes analyses suggested a predominant cochlear involvement. We conclude that hearing loss involving all frequency regions significantly contributes to morbidity in patients with Fabry disease. Our quantitative analysis suggests a correlation of neuropathic and vascular damage with hearing loss in the males. Residual GALA activity appears to have a protective effect against hearing loss.
Asunto(s)
Enfermedad de Fabry/fisiopatología , Pérdida Auditiva/fisiopatología , Adolescente , Adulto , Anciano , Niño , Estudios Transversales , Oído Medio/fisiopatología , Enfermedad de Fabry/complicaciones , Femenino , Pérdida Auditiva/complicaciones , Humanos , Lenguaje , Masculino , Microcirculación , Persona de Mediana Edad , Enfermedades del Sistema Nervioso Periférico/complicaciones , Enfermedades del Sistema Nervioso Periférico/fisiopatología , Pruebas Psicológicas , Estudios Retrospectivos , Umbral Sensorial , Índice de Severidad de la Enfermedad , Factores Sexuales , Telencéfalo/irrigación sanguínea , Acúfeno/complicaciones , Acúfeno/fisiopatología , alfa-Galactosidasa/metabolismoRESUMEN
Gaucher disease, the most common sphingolipidosis, is caused by a decreased activity of glucosylceramide beta-glucosidase, resulting in the accumulation of glucosylceramide in macrophage-derived cells known as Gaucher cells. Much of the storage material is thought to originate from the turnover of cell membranes, such as phagocytosed red and white blood cells. In this study, an in vitro model of Gaucher disease was developed by treating the murine macrophage cell line J774 with a specific inhibitor of glucosylceramide beta-glucosidase, conduritol B-epoxide, and feeding red blood cell ghosts, in order to mimic the disease state. It was found in this model system that glucosylceramide beta-glucosidase activity could be reduced to about 11-15% of the normal control level before increased storage of glucosylceramide occurred. This in vitro system allows insight into the correlation between enzyme activity and lipid storage as predicted by the theory of residual enzyme activity that was proposed by Conzelmann and Sandhoff.
Asunto(s)
Enfermedad de Gaucher/diagnóstico , Enfermedad de Gaucher/metabolismo , Glucosilceramidasa/metabolismo , Inositol/análogos & derivados , Animales , Técnicas de Cultivo de Célula/métodos , Línea Celular , Cromatografía Líquida de Alta Presión , Relación Dosis-Respuesta a Droga , Inhibidores Enzimáticos/farmacología , Membrana Eritrocítica/metabolismo , Enfermedad de Gaucher/genética , Glucosilceramidasa/antagonistas & inhibidores , Glucosilceramidas/metabolismo , Heterocigoto , Inositol/farmacología , Metabolismo de los Lípidos , Macrófagos/metabolismo , Ratones , Microscopía ElectrónicaRESUMEN
Patients with Gaucher disease have been classified as type 1 nonneuronopathic, type 2 acute neuronopathic, and type 3 chronic neuronopathic phenotypes. Increased quantities of glucocerebroside and glucosylsphingosine (glucopsychosine) are present in the brain of type 2 and type 3 Gaucher patients. Galactosylsphingosine has previously been shown to be neurotoxic in globoid cell leukodystrophy (Krabbe disease). To determine whether glucosylsphingosine is also neurotoxic, we examined its effect on cultured cholinergic neuron-like LA-N-2 cells. When these cells were exposed to 1, 5, or 10 microM glucosylsphingosine for a period of 18 h, they became shriveled, neurite outgrowth was suppressed, and the activities of the lysosomal enzymes glucocerebrosidase, sphingomyelinase, and beta-galactosidase were reduced in a dose-dependent manner. Acetylcholine in cells exposed to glucosylsphingosine also declined. Cells switched to glucosylsphingosine-free medium partially recovered. The data suggest that accumulation of glucosylsphingosine contributes to neuronal dysfunction and destruction in patients with neuronopathic Gaucher disease.
Asunto(s)
Encéfalo/metabolismo , Enfermedad de Gaucher/metabolismo , Degeneración Nerviosa/metabolismo , Neuronas/metabolismo , Esfingosina/análogos & derivados , Esfingosina/metabolismo , Acetilcolina/metabolismo , Encéfalo/patología , Encéfalo/fisiopatología , Línea Celular , Fibras Colinérgicas/efectos de los fármacos , Fibras Colinérgicas/metabolismo , Fibras Colinérgicas/patología , Medios de Cultivo/farmacología , Relación Dosis-Respuesta a Droga , Enfermedad de Gaucher/patología , Enfermedad de Gaucher/fisiopatología , Glucosilceramidasa/metabolismo , Humanos , Modelos Biológicos , Degeneración Nerviosa/inducido químicamente , Degeneración Nerviosa/patología , Neuritas/efectos de los fármacos , Neuritas/metabolismo , Neuritas/patología , Neuronas/efectos de los fármacos , Neuronas/patología , Psicosina/análogos & derivados , Esfingomielina Fosfodiesterasa/metabolismo , Esfingosina/toxicidad , beta-Galactosidasa/metabolismoRESUMEN
UNLABELLED: Over the past 40 years there has been remarkable development in our understanding of the pathophysiology of lysosomal storage disorders. This review describes the research carried out on the sphingolipid storage disorders from the first demonstration of the underlying metabolic abnormality in Gaucher disease to the development of enzyme replacement therapy for Gaucher and Fabry diseases. Initial developments in gene therapy are also described. CONCLUSION: The introduction of enzyme replacement therapy has provided a lifeline for patients with Gaucher or Fabry disease. It is anticipated that future developments, including gene therapy, will provide additional therapeutic options.
Asunto(s)
Enfermedad de Fabry/fisiopatología , Enfermedad de Fabry/terapia , Enfermedad de Gaucher/fisiopatología , Enfermedad de Gaucher/terapia , Terapia Genética , Glucosilceramidasa/administración & dosificación , Glucosilceramidasa/metabolismo , Glucosilceramidasa/uso terapéutico , Humanos , Proteínas Estructurales Virales , alfa-Galactosidasa/administración & dosificación , alfa-Galactosidasa/metabolismo , alfa-Galactosidasa/uso terapéuticoRESUMEN
Enzyme replacement therapy has been shown to be particularly effective for patients with type 1 (non-neuronopathic) Gaucher disease. However, intravenously administered glucocerebrosidase does not reverse or halt the progression of brain damage in patients with type 2 (acute neuronopathic) Gaucher disease. A previous investigation revealed that intracerebral infusion of mannose-terminal glucocerebrosidase was safe in experimental animals. The enzyme had a comparatively long half-life in the brain. It was transported by convection from the site of infusion along white matter fiber tracts to the cerebral cortex where it was endocytosed by neurons. In anticipation of intracerebral administration of mannose-terminal glucocerebrosidase to patients with type 2 Gaucher disease, it was important to learn the mechanism involved in its cellular uptake. We therefore compared the endocytosis of this enzyme by J774 macrophage cells with that in two human neuronal cell lines and a human astrocyte cell line. Mannose-terminal glucocerebrosidase was taken up by cholinergic LA-N-2 cells, but to a much lower extent than by macrophages. Considerably less of the enzyme was endocytosed by dopaminergic SH-SY5Y cells. It was not taken up by NHA astrocytes. The findings provide encouragement for an exploration of intracerebral administration of glucocerebrosidase in patients with type 2 Gaucher disease.
Asunto(s)
Acetilcolina/metabolismo , Dopamina beta-Hidroxilasa/metabolismo , Glucosilceramidasa/metabolismo , Neuronas/enzimología , Transporte Biológico , Diferenciación Celular , Línea Celular , Células Cultivadas , Medios de Cultivo , Enfermedad de Gaucher/enzimología , Humanos , Cinética , Manosa , Neuronas/citologíaRESUMEN
Fabry disease is an X-linked lysosomal disorder characterized by deficient alpha-galactosidase A activity and intracellular accumulations of glycosphingolipids, mainly globotriaosylceramide (Gb3). Clinically, patients occasionally present CNS dysfunction. To examine the pathophysiology underlying brain dysfunction, we examined glucose utilization (CMR(glc)) and cerebral blood flow (CBF) globally and locally in 18 brain structures in the alpha-galactosidase A gene knockout mouse. Global CMR(glc) was statistically significantly reduced by 22% in Fabry mice (p < 0.01). All 18 structures showed decreases in local CMR(glc) ranging from 14% to 33%. The decreases in all structures of the diencephalon, caudate-putamen, brain stem, and cerebellar cortex were statistically significant (p < 0.05). Global cerebral blood flow (CBF) and local CBF measured in the same 18 structures were lower in Fabry mice than in control mice, but none statistically significantly. Histological examination of brain revealed no cerebral infarcts but abundant Gb3 deposits in the walls of the cerebral vessels with neuronal deposits localized to the medulla oblongata. These results indicate an impairment in cerebral energy metabolism in the Fabry mice, but one not necessarily due to circulatory insufficiency.
Asunto(s)
Encéfalo/metabolismo , Circulación Cerebrovascular , Enfermedad de Fabry , Enfermedad de Fabry/metabolismo , Glucosa/metabolismo , Proteínas del Tejido Nervioso/deficiencia , Animales , Encéfalo/patología , Metabolismo Energético , Enfermedad de Fabry/genética , Enfermedad de Fabry/patología , Glicoesfingolípidos/metabolismo , Riñón/metabolismo , Hígado/metabolismo , Masculino , Bulbo Raquídeo/patología , Ratones , Ratones Noqueados , Modelos Animales , Músculo Liso Vascular/metabolismo , Músculo Liso Vascular/patología , Proteínas del Tejido Nervioso/genética , Neuronas/metabolismo , Neuronas/patología , Trihexosilceramidas/metabolismo , alfa-Galactosidasa/genéticaAsunto(s)
Enfermedad de Tay-Sachs/historia , beta-N-Acetilhexosaminidasas/historia , Encéfalo/enzimología , Enfermedad de Gaucher/enzimología , Enfermedad de Gaucher/genética , Enfermedad de Gaucher/historia , Historia del Siglo XX , Humanos , Enfermedad de Tay-Sachs/diagnóstico , Enfermedad de Tay-Sachs/enzimología , beta-N-Acetilhexosaminidasas/análisis , beta-N-Acetilhexosaminidasas/metabolismo , beta-N-Acetilhexosaminidasas/uso terapéuticoRESUMEN
The beneficial effects of macrophage-targeted glucocerebrosidase (alglucerase, Ceredase) in patients with Gaucher disease are well established. A minority of recipients develop transient non-neutralizing antibodies to the exogenous enzyme. A 7-year-old patient with type 3 Gaucher disease, whose clinical course began to deteriorate while receiving alglucerase developed a progressively increasing titer of IgG antibody, that blocked the catalytic activity of alglucerase. We investigated the acid beta-glucosidase genotype in this patient. Direct sequencing of both cDNA and genomic PCR products was used to characterize the mutations underlying acid beta-glucosidase deficiency. The patient was shown to be a compound heterozygote for a previously reported missense mutation (G377S), and a novel single nucleotide deletion (g5255delT). The transcript originating from the latter allele was undetectable in RT-PCR experiments. We report the first characterization of a GBA genotype associated with the development of neutralizing antibody to alglucerase, in a patient affected with type 3 Gaucher disease. Our results may help to shed light on the mechanisms underlying this phenomenon which, in the rare instances where it occurs, hampers the efficacy of enzyme replacement therapy.
Asunto(s)
Enfermedad de Gaucher/enzimología , Enfermedad de Gaucher/genética , Glucosilceramidasa/efectos adversos , Glucosilceramidasa/inmunología , Mutación , beta-Glucosidasa/genética , Formación de Anticuerpos , Secuencia de Bases , Niño , ADN/genética , Análisis Mutacional de ADN , Enfermedad de Gaucher/tratamiento farmacológico , Enfermedad de Gaucher/inmunología , Glucosilceramidasa/uso terapéutico , Heterocigoto , Humanos , Masculino , Mutación Missense , Pruebas de Neutralización , Eliminación de SecuenciaRESUMEN
Members of the N-methyl-d-aspartate (NMDA) class of glutamate receptors (NMDARs) are critical for development, synaptic transmission, learning and memory; they are targets of pathological disorders in the central nervous system. NMDARs are phosphorylated by both serine/threonine and tyrosine kinases. Here, we demonstrate that cyclin dependent kinase-5 (Cdk5) associates with and phosphorylates NR2A subunits at Ser-1232 in vitro and in intact cells. Moreover, we show that roscovitine, a selective Cdk5 inhibitor, blocks both long-term potentiation induction and NMDA-evoked currents in rat CA1 hippocampal neurons. These results suggest that Cdk5 plays a key role in synaptic transmission and plasticity through its up-regulation of NMDARs.
Asunto(s)
Quinasas Ciclina-Dependientes/fisiología , Receptores de N-Metil-D-Aspartato/metabolismo , Animales , Células Cultivadas , Quinasa 5 Dependiente de la Ciclina , Hipocampo/fisiología , Potenciación a Largo Plazo , Masculino , Ratones , Ratones Noqueados , Fosforilación , Ratas , Ratas Sprague-Dawley , Receptores de N-Metil-D-Aspartato/análisis , Serina/metabolismo , Transmisión SinápticaRESUMEN
Disorders that arise as a result of lysosomal dysfunction represent some of the most challenging diagnostic problems in medicine. Not only are these disorders infrequently seen, but they may also present with signs and symptoms that mimic perinatal injury, food intolerance, or the sequellae of neonatal infection. Misidentification can lead to significant delay in diagnosis. Ironically, as the prevailing economic climate places increasing time constraints on practicing physicians, medical research is providing treatment strategies and management techniques that are most effective if applied early in the course of the disease. Most lysosomal storage disorders can now be definitively diagnosed once the signs are recognized. In many cases the benefits of early diagnosis, enlightened management, and appropriate referral are considerable. The aim of this paper is to demystify this elusive class of diseases, to promote clinical vigilance in their detection, and to provide a systematic approach to diagnosis when clinical suspicion is aroused.
Asunto(s)
Enfermedades por Almacenamiento Lisosomal/diagnóstico , Gangliósidos/metabolismo , Glicosaminoglicanos/metabolismo , Humanos , Enfermedades por Almacenamiento Lisosomal/genética , Enfermedades por Almacenamiento Lisosomal/metabolismo , Lisosomas/metabolismo , FenotipoRESUMEN
BACKGROUND: Fabry disease is an X-linked lysosomal deficiency of alpha-galactosidase A that results in cellular accumulation of galacto-conjugates such as globotriosylceramide, particularly in blood vessels. It is associated with early-onset stroke and kidney and heart failure. METHODS AND RESULTS: Using [(15)O] H(2)O and PET, we found increased resting regional cerebral blood flow in Fabry disease without evidence of occlusive vasculopathy or cerebral hypoperfusion. Because nitric oxide is known to play an important role in vascular tone and reactivity, we studied plasma nitrate, nitrite, and low-molecular-weight S-nitrosothiol levels by chemiluminescence. Skin biopsy specimens and archived brain tissue were also examined immunohistochemically for nitrotyrosine. Plasma nitrate, nitrite, and low-molecular-weight S-nitrosothiol were in the normal range; however, enhanced nitrotyrosine staining was observed in dermal and cerebral blood vessels. After a double-blind, placebo-controlled trial of alpha-galactosidase A therapy, the resting regional cerebral blood flow in the treated group was significantly reduced, with a notable decrease of nitrotyrosine staining in dermal blood vessels. CONCLUSIONS: These findings suggest a chronic alteration of the nitric oxide pathway in Fabry disease, with critical protein nitration that is reversible with enzyme replacement therapy.
Asunto(s)
Corteza Cerebral/irrigación sanguínea , Trastornos Cerebrovasculares/prevención & control , Enfermedad de Fabry/tratamiento farmacológico , Óxido Nítrico/metabolismo , alfa-Galactosidasa/uso terapéutico , Adulto , Trastornos Cerebrovasculares/etiología , Trastornos Cerebrovasculares/metabolismo , Trastornos Cerebrovasculares/fisiopatología , Método Doble Ciego , Enfermedad de Fabry/complicaciones , Enfermedad de Fabry/metabolismo , Humanos , Masculino , Persona de Mediana Edad , Flujo Sanguíneo RegionalRESUMEN
CONTEXT: Fabry disease is a metabolic disorder without a specific treatment, caused by a deficiency of the lysosomal enzyme alpha-galactosidase A (alpha-gal A). Most patients experience debilitating neuropathic pain and premature mortality because of renal failure, cardiovascular disease, or cerebrovascular disease. OBJECTIVE: To evaluate the safety and efficacy of intravenous alpha-gal A for Fabry disease. DESIGN AND SETTING: Double-blind placebo-controlled trial conducted from December 1998 to August 1999 at the Clinical Research Center of the National Institutes of Health. PATIENTS: Twenty-six hemizygous male patients, aged 18 years or older, with Fabry disease that was confirmed by alpha-gal A assay. INTERVENTION: A dosage of 0.2 mg/kg of alpha-gal A, administered intravenously every other week (12 doses total). MAIN OUTCOME MEASURE: Effect of therapy on neuropathic pain while without neuropathic pain medications measured by question 3 of the Brief Pain Inventory (BPI). RESULTS: Mean (SE) BPI neuropathic pain severity score declined from 6.2 (0.46) to 4.3 (0.73) in patients treated with alpha-gal A vs no significant change in the placebo group (P =.02). Pain-related quality of life declined from 3.2 (0.55) to 2.1 (0.56) for patients receiving alpha-gal A vs 4.8 (0.59) to 4.2 (0.74) for placebo (P =.05). In the kidney, glomeruli with mesangial widening decreased by a mean of 12.5% for patients receiving alpha-gal vs a 16.5% increase for placebo (P =.01). Mean inulin clearance decreased by 6.2 mL/min for patients receiving alpha-gal A vs 19.5 mL/min for placebo (P =.19). Mean creatinine clearance increased by 2.1 mL/min (0.4 mL/s) for patients receiving alpha-gal A vs a decrease of 16.1 mL/min (0.3 mL/s) for placebo (P =.02). In patients treated with alpha-gal A, there was an approximately 50% reduction in plasma glycosphingolipid levels, a significant improvement in cardiac conduction, and a significant increase in body weight. CONCLUSION: Intravenous infusions of alpha-gal A are safe and have widespread therapeutic efficacy in Fabry disease.
Asunto(s)
Enfermedad de Fabry/tratamiento farmacológico , alfa-Galactosidasa/uso terapéutico , Adulto , Análisis de Varianza , Arritmias Cardíacas , Peso Corporal , Método Doble Ciego , Esquema de Medicación , Enfermedad de Fabry/fisiopatología , Frecuencia Cardíaca , Humanos , Infusiones Intravenosas , Pruebas de Función Renal , Masculino , Dimensión del Dolor , Trihexosilceramidas/metabolismo , alfa-Galactosidasa/administración & dosificaciónRESUMEN
OBJECTIVE: To assess the long-term systemic and neurologic responses to enzyme replacement therapy (ERT) with macrophage-targeted glucocerebrosidase in patients with type 3 Gaucher's disease. STUDY DESIGN: Patients with type 3 Gaucher's disease (n = 21), aged 8 months to 35 years, were enrolled in a prospective study. Enzyme dose was adjusted to control systemic manifestations. Clinical and laboratory evaluations were performed at baseline and every 6 to 12 months thereafter. Patients were followed up for 2 to 8 years. RESULTS: Significant improvement in hemoglobin levels, platelet count, and acid phosphatase values occurred. Liver and spleen volume markedly decreased, and bone structure improved. Nineteen patients had asymptomatic interstitial lung disease unresponsive to ERT. Supranuclear gaze palsy remained stable in 19 patients, worsened in one patient, and improved in one. Cognitive function remained unchanged or improved over time in 13 patients but decreased in 8 patients, 3 of whom developed progressive myoclonic encephalopathy accompanied by cranial magnetic resonance imaging and electroencephalographic deterioration. CONCLUSIONS: At relatively high doses, ERT reverses almost all the systemic manifestations in patients with type 3 Gaucher's disease. Most treated patients do not deteriorate neurologically. Novel therapeutic strategies are required to reverse the pulmonary and neuronopathic aspects of the disease.
Asunto(s)
Enfermedad de Gaucher/tratamiento farmacológico , Glucosilceramidasa/uso terapéutico , Adolescente , Adulto , Determinación de la Edad por el Esqueleto , Niño , Preescolar , Electroencefalografía/métodos , Femenino , Enfermedad de Gaucher/sangre , Enfermedad de Gaucher/diagnóstico , Enfermedad de Gaucher/psicología , Glucosilceramidasa/administración & dosificación , Humanos , Pruebas de Inteligencia , Imagen por Resonancia Magnética , Masculino , Tomografía Computarizada por Rayos X , Resultado del TratamientoRESUMEN
Fabry disease is a lipid storage disorder resulting from mutations in the gene encoding the enzyme alpha-galactosidase A (alpha-gal A; EC ). We previously have demonstrated long-term alpha-gal A enzyme correction and lipid reduction mediated by therapeutic ex vivo transduction and transplantation of hematopoietic cells in a mouse model of Fabry disease. We now report marked improvement in the efficiency of this gene-therapy approach. For this study we used a novel bicistronic retroviral vector that engineers expression of both the therapeutic alpha-gal A gene and the human IL-2Ralpha chain (huCD25) gene as a selectable marker. Coexpression of huCD25 allowed selective immunoenrichment (preselection) of a variety of transduced human and murine cells, resulting in enhanced intracellular and secreted alpha-gal A enzyme activities. Of particular significance for clinical applicability, mobilized CD34(+) peripheral blood hematopoietic stem/progenitor cells from Fabry patients have low-background huCD25 expression and could be enriched effectively after ex vivo transduction, resulting in increased alpha-gal A activity. We evaluated effects of preselection in the mouse model of Fabry disease. Preselection of transduced Fabry mouse bone marrow cells elevated the level of multilineage gene-corrected hematopoietic cells in the circulation of transplanted animals and improved in vivo enzymatic activity levels in plasma and organs for more than 6 months after both primary and secondary transplantation. These studies demonstrate the potential of using a huCD25-based preselection strategy to enhance the clinical utility of ex vivo hematopoietic stem/progenitor cell gene therapy of Fabry disease and other disorders.
Asunto(s)
Enfermedad de Fabry/terapia , Terapia Genética/métodos , Receptores de Interleucina-2/genética , alfa-Galactosidasa/genética , Células 3T3 , Animales , Trasplante de Médula Ósea , Modelos Animales de Enfermedad , Enfermedad de Fabry/metabolismo , Enfermedad de Fabry/patología , Expresión Génica , Vectores Genéticos , Células HeLa , Humanos , Ratones , Ratones Endogámicos C57BL , Receptores de Interleucina-2/metabolismo , Retroviridae/genética , alfa-Galactosidasa/metabolismoRESUMEN
Cyclin-dependent kinase 5 (Cdk5) null mice exhibit a unique phenotype characterized by perinatal mortality, disrupted cerebral cortical layering attributable to abnormal neuronal migration, lack of cerebellar foliation, and chromatolytic changes of neurons in the brainstem and the spinal cord. Because Cdk5 is expressed in both neurons and astrocytes, it has been unclear whether this phenotype is primarily attributable to defects in neurons or in astrocytes. Herein we report reconstitution of Cdk5 expression in neurons in Cdk5 null mice and its effect on the null phenotype. Unlike the Cdk5 null mice, the reconstituted Cdk5 null mice that express the Cdk5 transgene under the p35 promoter (TgKO mice) were viable and fertile. Because Cdk5 expression is mainly limited to neurons in these mice and rescues the defects in the nervous system of the Cdk5 null phenotype, it clearly demonstrates that Cdk5 activity is necessary for normal development and survival of p35-expressing neurons.