RESUMO
DNA-microarrays allow the analysis of almost the complete gene expression program of tumor samples and normal control samples in a single experiment. This allows the processing of a large number of samples in a reasonable short time. Tumor specific gene expression profiles can be used for molecular tumor classification and as a new diagnostic tool. In addition, the identification of tumor specific genes can help to understand the biology of tumor cells and identified genes can be used for the development of new therapeutic strategies. However, the huge amount of data generated by DNA-microarrays creates new challenges for data analysis. In addition, accuracy and reproducibility of the available techniques require complementary methods for verification of DNA-microarray data.
Assuntos
Perfilação da Expressão Gênica , Proteínas de Neoplasias/genética , Neoplasias/genética , Análise de Sequência com Séries de Oligonucleotídeos , Criança , Regulação Neoplásica da Expressão Gênica/fisiologia , Terapia Genética , Humanos , Imunoterapia , Neoplasias/diagnóstico , Neoplasias/terapia , Valor Preditivo dos TestesRESUMO
Deficiency of the granulocyte-macrophage colony-stimulating factor (GM-CSF)/interleukin-3 (IL-3)/IL-5 receptors common beta chain (betac) is a cause of fatal respiratory failure. betac deficiency manifests as pulmonary alveolar proteinosis (PAP). PAP has heterogenous etiologies that may be genetic or aquired. Some cases of PAP have been reported to be associated with hematologic malignancies such as acute myeloid leukemia (AML). In mice, the PAP phenotype was generated by targeted deletion of the gene for betac and can be treated by transplantation of wild-type bone marrow into betac -/- mice. Thus, our findings in betac -/- mice provide evidence for a causal relationship between the lung disease and the hematopoietic system. We describe here expression defects of betac or betac plus GM-CSF receptor alpha chain (GM-CSFR alpha) in 3 pediatric patients with AML and PAP symptoms. All of the patients' leukemic cells failed to express normal levels of betac. The leukemic cells of patients no. 2 and 3 additionally lacked the expression of GM-CSFR alpha, as shown by flow cytometry. Strikingly reduced or absent function of betac was demonstrated in clonogenic progenitor assays with absent colony-forming unit (CFU) growth after GM-CSF or IL-3 stimulation. The response to growth factors acting via a growth factor receptor distinct from the GM-CSF/IL-3/IL-5 system (recombinant human granulocyte colony-stimulating factor [rhG-CSF]) was normal. After antileukemic treatment, the pulmonary symptoms resolved and betac or betac plus GM-CSFR alpha expression was normal. Our findings provide evidence that a defect in the expression of betac or betac plus GM-CSFR alpha on AML blasts can be associated with respiratory failure in patients with AML.
Assuntos
Regulação Leucêmica da Expressão Gênica , Leucemia Mieloide/complicações , Proteinose Alveolar Pulmonar/etiologia , Receptores de Fator Estimulador das Colônias de Granulócitos e Macrófagos/deficiência , Receptores de Interleucina-3/deficiência , Receptores de Interleucina/deficiência , Insuficiência Respiratória/etiologia , Doença Aguda , Adulto , Animais , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Transplante de Medula Óssea , Líquido da Lavagem Broncoalveolar/química , Líquido da Lavagem Broncoalveolar/citologia , Linhagem da Célula , Criança , Evolução Fatal , Feminino , Humanos , Lactente , Leucemia Monocítica Aguda/complicações , Leucemia Monocítica Aguda/genética , Leucemia Monocítica Aguda/terapia , Leucemia Mieloide/tratamento farmacológico , Leucemia Mieloide/genética , Leucemia Mielomonocítica Aguda/complicações , Leucemia Mielomonocítica Aguda/tratamento farmacológico , Leucemia Mielomonocítica Aguda/genética , Macrófagos Alveolares/patologia , Masculino , Camundongos , Camundongos Knockout , Proteinose Alveolar Pulmonar/diagnóstico , Proteinose Alveolar Pulmonar/genética , Surfactantes Pulmonares/análise , Receptores de Fator Estimulador das Colônias de Granulócitos e Macrófagos/biossíntese , Receptores de Fator Estimulador das Colônias de Granulócitos e Macrófagos/química , Receptores de Fator Estimulador das Colônias de Granulócitos e Macrófagos/genética , Receptores de Interleucina/biossíntese , Receptores de Interleucina/química , Receptores de Interleucina/genética , Receptores de Interleucina-3/biossíntese , Receptores de Interleucina-3/química , Receptores de Interleucina-3/genética , Receptores de Interleucina-5RESUMO
Pulmonary alveolar proteinosis (PAP) is a heterogeneous disorder of genetic or acquired etiologies. In some cases congenital PAP is associated with hereditary surfactant protein (SP)-B deficiency. To date, the molecular defect in the majority of patients with PAP has not been identified. In mice, PAP has been generated by targeted deletion of the genes for either the GM-CSF/IL-3/IL-5 receptor common beta chain (beta c) or GM-CSF. Here, we describe an expression defect of beta c in three of seven pediatric patients with PAP and in one patient with severe lung disease suspected to be PAP. The patients failed to express normal levels of beta c as shown by flow cytometry. Strikingly reduced or absent function of beta c was demonstrated by ligand binding studies and progenitor clonogenic assays. Analysis of beta c DNA revealed a point mutation from proline to threonine at codon 602 in one patient. Our findings provide evidence that a defect in the expression of a hematopoietic cytokine receptor is associated with human PAP.
Assuntos
Proteinose Alveolar Pulmonar/metabolismo , Receptores de Fator Estimulador das Colônias de Granulócitos e Macrófagos/deficiência , Receptores de Interleucina-3/deficiência , Receptores de Interleucina/deficiência , Adulto , Substituição de Aminoácidos , Pré-Escolar , Fator Estimulador de Colônias de Granulócitos e Macrófagos/metabolismo , Hematopoese , Humanos , Lactente , Cinética , Mutação Puntual , Proteolipídeos/metabolismo , Surfactantes Pulmonares/metabolismo , Receptores de Interleucina-5RESUMO
We report the DNA sequence of a 34,038 bp segment of Saccharomyces cerevisiae chromosome XV. Subsequent analysis revealed 20 open reading frames (ORFs) longer than 300 bp and two tRNA genes. Five ORFs correspond to genes previously identified in S. cerevisiae, including RPLA2, PRE6, MSE1, IFM1 and SCM2 (TAT2, TAP2, LTG3). Two putative proteins share considerable homology with other proteins in the current data libraries. ORF O2145 shows 41.2% identity with the glycophospholipid-anchored surface glycoprotein Gas1p of S. cerevisiae and ORF O2197 has 53.2% identity to chromosome segregation protein Dis3p of Schizosaccharomyces pombe.