Pathologic gene expression in Gaucher disease: up-regulation of cysteine proteinases including osteoclastic cathepsin K. (25/509)

Deficiency of lysosomal acid beta-glucosidase induces glycolipid storage in the macrophages of Gaucher disease but the pathways of multisystem tissue injury and destruction are unknown. To investigate the cognate molecular pathology of this inflammatory disorder, genes that were differentially expressed in spleen samples from a patient with Gaucher disease (Gaucher spleen) were isolated. Of 64 complementary DNA (cDNA) fragments sequenced from an enriched Gaucher cDNA library, 5 encode lysosomal proteins (cathepsins B, K, and S, alpha-fucosidase, and acid lipase), 10 encode other known proteins, and 2 represent novel sequences from human macrophage cell lines. Transcript abundance of the cathepsins, novel genes, pulmonary and activation-regulated chemokine (PARC), and NMB, a putative tumor suppressor gene, was greatly increased. Immunoblotting showed increased mature forms of all 3 cathepsins found in samples of Gaucher spleens. Immunofluorescence microscopy showed strong cathepsin B and K reactions in sinusoidal endothelium and Gaucher cells. The respective means, plus or minus SD, of cathepsin B, K, and S activities were 183 +/- 35, 97 +/- 39, and 91 +/- 45 nmol/min/mg protein in 4 Gaucher spleens, and 26 +/- 4, 10.5 +/- 2, and 4.0 +/- 2.1 nmol/min/mg protein in 3 control spleens. Plasma cathepsin B, K, and S activities were also elevated in Gaucher disease plasma (P <.001), but compared with control plasma samples, neither cathepsin B nor K activities were significantly elevated in 8 patients with nonglycosphingolipid lysosomal storage diseases or in 9 patients with other glycosphingolipidoses, which suggests disease specificity. All 3 cathepsin activities were increased 2-fold to 3-fold in Gaucher sera compared with control sera. In all 6 patients treated by enzyme replacement for 16-22 months, serum cathepsin activities decreased significantly (P <.01). Longitudinal studies confirmed the progressive reduction of proteinase activities during imiglucerase therapy but in 3 Gaucher patients with mild disease not so treated, serum cathepsin activities remained constant or increased during follow-up. Enhanced expression of cysteine proteinases may promote tissue destruction. Moreover, the first identification of aberrant cathepsin K expression in hematopoietic tissue other than osteoclasts implicates this protease in the breakdown of the matrix that characterizes lytic bone lesions in Gaucher disease. (Blood. 2000;96:1969-1978)  (+info)

Longterm follow-up of electroencephalographic and clinical findings of a case with Gaucher's disease type 3a. (26/509)

Among three recognised clinical phenotypes, type 3a Gaucher's disease is characterised by mild to severe systemic disease, neurological manifestations and myoclonic seizures. We report the long term clinical and electrophysiological follow-up of a 27-year old man with a diagnosis of type 3a Gaucher's disease, which was confirmed by bone marrow biopsy examination and leukocyte glucocerebrosidase level measurement. His neurological examination was normal throughout the follow-up period. EEG examination, recorded five days after the first seizure, revealed generalised nonrhythmic paroxysmal rapid spikes with occipital predominance increased by photic stimulation and normal background activity. The frequency of seizures increased from 3-4/year to 1-2/month within a follow-up period of 12 years and a repeat EEG examination on the eight year of diagnosis revealed additional background slowing. A giant potential was obtained in somatosensory evoked potential (SEP) examination. EEG findings of this case demonstrate a specific pattern with rapid spike activity, photosensitivity, eye closure sensitivity and gradual background slowing.  (+info)

Metabolism of glucosyl [13H]ceramide by human skin fibroblasts from normal and glucosylceramidotic subjects. (27/509)

Metabolic utilization of glucosyl [3H]ceramide (glucocerebroside) by human skin fibroblasts from normal and glucosylceramidotic subjects was examined in cell culture. Exogenous glucosyl [3H]ceramide in the culture medium did not influence activity of "acid" beta glucosidase in either cell type. Expectedly in the lipidotic cells, the enzymatic activity was markedly (similar to 20-fold) lower. Normal cells were found preferentially to utilize exogenous tritium-labeled glucosylceramide for a source of precursors for phopholipid biosynthesis. The fatty acid and the sphingosine components of sphingomyelin, and the fatty acid components of phosphatidylcholine and phosphatidylethanolamine, but not phosphatidylserine and phosphatidylinositol, were tritium-labeled. In contrast, glucosylceramidotic cells utilized labeled glucosylceramide far more for synthesis of lactosylceramide (lactocerebroside) and also higher neutral glycosphingolipid homologues. These experimental findings suggest that glucosyl [3H]ceramide is hydrolyzed in normal skin fibroblasts to [3H]ceramide and further to [3H]dihydrosphingosine and 3H-labeled fatty acids. These compounds are subsequently incorporated into cellular phospholipids. Flux of glucosyl [3H]ceramide through this catabolic sequence and reincorporation of its breakdown products into phospholipids predominates in normal skin fibroblasts. In contrast, it is greatly reduced in glucosylceramidotic skin fibroblasts. Consequently, a greater amount of glucosyl [3H]ceramide remains intact for the synthesis of more highly glycosylated glycosphingolipids in the latter cells.  (+info)

The facile detection of 1505G-->A in Gaucher patients with different phenotypes. (28/509)

In Gaucher disease patients, over 100 disease-causing mutations have been identified. For identification of the 1504C-->T (R463C) mutation it is common to use PCR-restriction fragmentation analysis using the restriction enzyme MspI. In the present study we investigated the reliability of this approach because accurate determination of genotypes is important in genotype-phenotype correlations. A simple modification, i.e. using the restriction enzyme HphI instead of MspI, revealed that type I and II Gaucher disease patients who had previously been identified as carrying the 1504C-->T mutation in fact carried the 1505G-->A (IVS10(-1)G-->A) mutation. Sequencing of the appropriate fragment confirmed this. The PCR method easily differentiates between these two mutations in Gaucher disease patients, thus circumventing the need for sequencing procedures. The phenotypes of the patients found to be carrying the 1505G-->A mutation are also described.  (+info)

Lung involvement and enzyme replacement therapy in Gaucher's disease. (29/509)

Symptomatic lung involvement in Gaucher's disease is relatively rare, being restricted to patients with other severe manifestations. We describe our experience in eight of 411 patients in our referral clinic, who presented with prominent pulmonary signs or symptoms. There were four adults and four children; all have been successfully treated with enzyme replacement therapy. Routine means of monitoring pulmonary status including clinical assessment, chest X-ray, pulmonary function tests, and high-resolution CT (HRCT) were used. Enzyme treatment resulted in decreased hepatosplenomegaly, improved haematological parameters, and increased well-being; There was decreased clubbing and decreased dyspnoea in some of the patients, although on radiology, lung pathology had not normalized. All four children showed improved respiratory compliance, with significant improvement of the radiological findings in one and unchanged disease in the others. Two adults showed improvement in oxygen saturation but worsening of pulmonary hypertension. On chest X-ray, both had increased interstitial markings; one had gradual progression of pulmonary artery accentuation and fine interstitial stable pattern on HRCT. The other two adults had no change in lung function or on chest X-ray, but on HRCT there was apparent improvement in one patient. There is great heterogeneity in presentation and response to enzyme therapy in patients with Gaucher's disease and symptomatic lung involvement. Clinically, some benefited significantly from enzyme therapy, but in contrast to the dramatic reduction in organomegaly, there was no normalization in pulmonary function or lung architecture.  (+info)

Initiation of enzyme replacement therapy for an adult patient with asymptomatic type 1 Gaucher's disease. (30/509)

A 27-year-old woman was admitted for further examination of thrombocytopenia. Symptoms were absent, but physical examination demonstrated hepatosplenomegaly without neurological abnormalities. Bone marrow examination revealed many Gaucher cells, and glucocerebrosidase activity from cultured skin fibroblasts was markedly reduced. A 1448C (L444P) mutation was detected on one allele of the glucocerebrosidase gene. Because magnetic resonance imaging (MRI) of the femora indicated severe infiltration of Gaucher cells into bone marrow, enzyme replacement therapy was initiated despite the absence of skeletal symptoms. Hematologic abnormalities, visceral and bone involvement have been improving. In cases of thrombocytopenia or hepatosplenomegaly, Gaucher's disease should be suspected.  (+info)

Chitotriosidase genotype and plasma activity in patients type 1 Gaucher's disease and their relatives (carriers and non carriers). (31/509)

BACKGROUND AND OBJECTIVES: Chitinases are enzymes that hydolyze chitin and have been found in a wide variety of nonvertebrate species; recently an human analogue of chitinases, chitotriosidase (CT) has been identified. Extreme elevations of plasma CT activity are observed in patients with Gaucher disease (GD), being Gaucher cells the source of the CT. It has been reported a 24 bp duplication in CT gene that results an inactive protein. The carrier prevalence is high as 30 to 40% and the CT activity is half that in wild individuals. However no systematic evaluation of plasma CT activity has been carried in GD patients taken into account status of allele defective for CT and dose in patients on enzyme replacement therapy (ERT). DESIGN AND METHODS: We had previously study 210 subjects from 99 unrelated Spanish GD families; 121 were non-affected carriers and 89 were non-carriers to establish carrier prevalence of CT genotypes. Plasma CTactivity and CTgenotypes by PCR and gel electrophoresis have been measured in 109 GD patients before treatment. We also evalued CT activity after ERT with alglucerase in 68 patients. RESULTS: Three patients had defective activities of CT. The carrier prevalence for 24 bp duplication was 35% and the allele frequency 0.20. No correlation between CT activity and GBA genotype was detected and between CT activity and visceral or skeletal disease in GD patients. Untreated affected patients, non-carriers for the duplication, had higher CT activity than carriers (p<.0001). CT activity decreased dramatically during the first 12 months of ERT; even after 3 years of therapy a persistent fall of CT activity was observed. How ever within 3 years of treatment, a significant difference in the mean decrease of CT activity was present among the groups of patients on varying alglucerase doses (p<.01). After 12 months of ERT the activity of plasma CT decline in the same percentage in both groups: heterozygous for the carriers of 24bp duplication and wild type, but thereafter CT activity decline more slowly in carriers than non carriers. INTERPRETATION AND CONCLUSIONS: The present data can be used as a reference to interpret the CT activity in GD patients with or without ERT, as well as to evaluate the doses response and can be used as a reference to interpret the CTactivity in carriers and non-carriers.  (+info)

Ionization and fragmentation of neutral and acidic glycosphingolipids with a Q-TOF mass spectrometer fitted with a MALDI ion source. (32/509)

This paper reports the use of a quadrupole time-of-flight (Q-TOF) mass spectrometer fitted with a matrix-assisted laser desorption/ionization (MALDI) ion source for the analysis of neutral and acidic glycosphingolipids. All compounds gave strong [M + Na]+ ions with 2,5-dihydroxybenzoic acid as the matrix, with no loss of sensitivity with increasing mass as was observed from the corresponding ions produced by electrospray. Neutral glycosphingolipids showed negligible in-source fragmentation but sialylated compounds fragmented by loss of sialic acid. However, these losses were not accompanied by unfocused post-source-decay ions as observed with MALDI-reflectron-TOF instruments. The MS/MS spectra were almost identical to those obtained by electrospray. Fragmentation of all compounds was mainly by glycosidic cleavage to give ions, both with and without the ceramide moiety, which defined the carbohydrate chain sequence. Weak ions which defined the sphingosine chain length and abundant ions, produced by loss of the acyl chain, were present when this chain contained a 2-hydroxy group. The technique was applied to the identification of ceramide-trihexosides present in tissues from mice genetically modified to model one of the glycolipid storage diseases (Fabry disease).  (+info)