RESUMO
Advances in the knowledge of the molecular genetics of Gaucher disease has made diagnosis more certain. Carrier detection in kindreds in which the responsible mutation has been defined is completely reliable now. Coupled with enzymatic assays, the diagnostic capabilities are greater than before. Use of these methods provides important information to individuals at risk and allows them to make critical decisions. The new, simplified methods reviewed in this article permit the molecular diagnosis of the disease and carrier stage of large numbers of samples within 1 week.
Assuntos
Doença de Gaucher/diagnóstico , Doença de Gaucher/metabolismo , Análise de Sequência de DNA/métodos , Sequência de Bases , Análise Mutacional de DNA/métodos , Eletroforese em Gel de Poliacrilamida , Aconselhamento Genético , Testes Genéticos , Genótipo , Glucosilceramidas/genética , Humanos , Biologia Molecular , Dados de Sequência Molecular , Fenótipo , Reação em Cadeia da PolimeraseRESUMO
A new test for the diagnosis of Gaucher disease is described. The test is designed to screen large numbers of clinical specimens from high-risk populations. It consists of duplex PCR amplification of genomic DNA followed by hybridization to alkaline phosphatase-conjugated allele-specific oligonucleotide probes (ASOs). High melting temperature PCR primers were used to increase specificity and eliminate the need for a separate annealing step. All hybridization and washing steps were performed at one temperature. Chemiluminescent detection of signals is fast, and results are easily interpreted directly from x-ray films. Currently, the test is being used in our laboratories to screen Ashkenazi Jewish populations in whom Gaucher disease is common.
Assuntos
Doença de Gaucher/diagnóstico , Reação em Cadeia da Polimerase , Doença de Gaucher/sangue , Doença de Gaucher/genética , Triagem de Portadores Genéticos , Testes Genéticos , Humanos , Judeus/genética , Mutação , Sondas de OligonucleotídeosRESUMO
This report summarizes the results on 39 patients with Gaucher disease who have been genotyped, evaluated, and/or followed at this center. Mutation analysis for 4 common mutations; N370S, L444P, 84gg and IVS2 (+1), was performed for all patients. Mutation analysis identified both mutant alleles in 69% and at least one mutant allele in 90% of all chromosomes. This study group of 39 patients included 32 type 1, four type 2 and three type 3 patients. We include the details of the clinical course of two patients with Gaucher disease treated with enzyme replacement therapy (ERT). One patient with chronic neuronopathic Gaucher disease has been treated with enzyme replacement therapy (ERT) at a dose of 60 U/kg every 2 weeks since 2.5 years of age and has shown no progression of neurologic involvement. A second patient with non-neuronopathic Gaucher disease has demonstrated an unusually delayed response to ERT. No clinical response was noted following 17 months of treatment at 60 U/kg every 2 weeks. Only after the dose was increased to 60 U/kg every week was a clinical response evident. Response to treatment at 15 U/kg every 2 weeks was variable in the four type 1 patients treated at the lower dose. In two of these patients with identical genotypes, one patient demonstrated a positive clinical response to low dose treatment while the other patient did not.
Assuntos
Doença de Gaucher/tratamento farmacológico , Doença de Gaucher/genética , Glucosilceramidase/genética , Glucosilceramidase/uso terapêutico , Adolescente , Adulto , Idoso , Sistema Nervoso Central/fisiopatologia , Criança , Pré-Escolar , Relação Dose-Resposta a Droga , Feminino , Doença de Gaucher/epidemiologia , Humanos , Lactente , Judeus , Masculino , Pessoa de Meia-Idade , Mutação , Baço/patologia , População BrancaRESUMO
The inherited deficiency of glucocerebrosidase results in a group of sphingolipid storage disorders referred to collectively as Gaucher disease. Study of the biochemistry and cell biology of glucocerebrosidase has made possible an effective enzyme replacement therapy for the disease. Definition of the molecular genetics of glucocerebrosidase has improved diagnostic capabilities and presents the exciting possibility of a cure through gene therapy.
Assuntos
Doença de Gaucher/terapia , Terapia Genética , Glucosilceramidase/genética , Animais , Animais Geneticamente Modificados , Sequência de Carboidratos , Análise Mutacional de DNA , Doença de Gaucher/diagnóstico , Doença de Gaucher/enzimologia , Técnicas de Transferência de Genes , Glucosilceramidase/biossíntese , Glucosilceramidase/química , Humanos , Dados de Sequência Molecular , Oligossacarídeos/química , Reação em Cadeia da PolimeraseRESUMO
Transfer of the gene coding for glucocerebrosidase (GC) via a retroviral vector (MFG-GC) to haematopoietic progenitors results in engraftment and life-long expression of the human protein at high levels in transplanted mice. Studies of human CD34 cells were carried out to evaluate their potential use in a gene therapy approach to Gaucher's disease. High transduction efficiency and correction of the enzyme deficiency was possible in CD34 cells obtained from patients with Gaucher's disease. Based on these results, a clinical trial of gene therapy was designed and initiated. Preliminary results of this study indicate the persistence or engraftment of genetically corrected cells in the transplanted patients.