RESUMEN
This work continues the series of the quantitative measurements of the proteins encoded by different chromosomes in the blood plasma of a healthy person. Selected Reaction Monitoring with Stable Isotope-labeled peptide Standards (SRM SIS) and a gene-centric approach, which is the basis for the implementation of the international Chromosome-centric Human Proteome Project (C-HPP), were applied for the quantitative measurement of proteins in human blood plasma. Analyses were carried out in the frame of C-HPP for each protein-coding gene of the four human chromosomes: 18, 13, Y, and mitochondrial. Concentrations of proteins encoded by 667 genes were measured in 54 blood plasma samples of the volunteers, whose health conditions were consistent with requirements for astronauts. The gene list included 276, 329, 47, and 15 genes of chromosomes 18, 13, Y, and the mitochondrial chromosome, respectively. This paper does not make claims about the detection of missing proteins. Only 205 proteins (30.7%) were detected in the samples. Of them, 84, 106, 10, and 5 belonged to chromosomes 18, 13, and Y and the mitochondrial chromosome, respectively. Each detected protein was found in at least one of the samples analyzed. The SRM SIS raw data are available in the ProteomeXchange repository (PXD004374, PASS01192).
Asunto(s)
Cromosomas Humanos/química , Plasma/química , Proteoma , Cromosomas Humanos/genética , Cromosomas Humanos Par 13/química , Cromosomas Humanos Par 18/química , Cromosomas Humanos Y/química , Bases de Datos de Proteínas , Voluntarios Sanos , Humanos , Mitocondrias/ultraestructura , Proteoma/genéticaRESUMEN
Extrachromosomal circular DNA (eccDNA) and ring chromosomes are genetic alterations found in humans with genetic disorders. However, there is a lack of genetic engineering tools to recapitulate and study the biogenesis of eccDNAs. Here, we created a dual-fluorescence biosensor cassette, which upon the delivery of pairs of CRISPR/Cas9 guide RNAs, CRISPR-C, allows us to study the biogenesis of a specific fluorophore expressing eccDNA in human cells. We show that CRISPR-C can generate functional eccDNA, using the novel eccDNA biosensor system. We further reveal that CRISPR-C also can generate eccDNAs from intergenic and genic loci in human embryonic kidney 293T cells and human mammary fibroblasts. EccDNAs mainly forms by end-joining mediated DNA-repair and we show that CRISPR-C is able to generate endogenous eccDNAs in sizes from a few hundred base pairs and ranging up to 207 kb. Even a 47.4 megabase-sized ring chromosome 18 can be created by CRISPR-C. Our study creates a new territory for CRISPR gene editing and highlights CRISPR-C as a useful tool for studying the cellular impact, persistence and function of eccDNAs.
Asunto(s)
Proteína 9 Asociada a CRISPR/genética , Sistemas CRISPR-Cas , Repeticiones Palindrómicas Cortas Agrupadas y Regularmente Espaciadas , ADN Circular/genética , Edición Génica/métodos , Secuencia de Bases , Técnicas Biosensibles , Proteína 9 Asociada a CRISPR/metabolismo , Línea Celular , Cromosomas Humanos Par 18/química , Cromosomas Humanos Par 18/metabolismo , Reparación del ADN por Unión de Extremidades , ADN Circular/metabolismo , Fibroblastos , Colorantes Fluorescentes/química , Colorantes Fluorescentes/metabolismo , Genes Reporteros , Sitios Genéticos , Vectores Genéticos/química , Vectores Genéticos/metabolismo , Genoma Humano , Proteínas Fluorescentes Verdes/genética , Proteínas Fluorescentes Verdes/metabolismo , Células HEK293 , Humanos , Proteínas Luminiscentes/genética , Proteínas Luminiscentes/metabolismo , ARN Guía de Kinetoplastida/genética , ARN Guía de Kinetoplastida/metabolismoRESUMEN
In the boundaries of the chromosome-centric Human Proteome Project (c-HPP) to obtain information about proteoforms coded by chromosome 18, several cell lines (HepG2, glioblastoma, LEH), normal liver, and plasma were analyzed. In our study, we have been using proteoform separation by two-dimensional electrophoresis (2DE) (a sectional analysis) and a semivirtual 2DE with following shotgun mass spectrometry using LC-ESI-MS/MS. Previously, we published a first draft of this research, where only HepG2 cells were tested. Here, we present the next step using more detailed analysis and more samples. Altogether, confident (2 significant sequences minimum) information about proteoforms of 117 isoforms coded by 104 genes of chromosome 18 was obtained. The 3D-graphs showing distribution of different proteoforms from the same gene in the 2D map were generated. Additionally, a semivirtual 2DE approach has allowed for detecting more proteoforms and estimating their pI more precisely. Data are available via ProteomeXchange with identifier PXD010142.
Asunto(s)
Cromosomas Humanos Par 18/química , Electroforesis en Gel Bidimensional/métodos , Isoformas de Proteínas/análisis , Proteoma/análisis , Línea Celular , Cromatografía Liquida , Simulación por Computador , Humanos , Proteómica/métodos , Espectrometría de Masas en TándemRESUMEN
Follicular lymphoma (FL) is generally considered an indolent disorder. With modern day treatments, long remissions are often achieved both in the front-line and relapsed setting. However, a subset of patients has a more aggressive course and a worse outcome. Their identification is the main purpose of modern day prognostic tools. In this review, we attempt to summarize the evidence concerning prognostic and predictive factors in FL, including (1) pre-treatment factors, from baseline clinical characteristics and imaging tests to histological grade, the microenvironment and genomic abnormalities; (2) post-treatment factors, i.e., depth of response, measured both by imaging tests and minimal residual disease; (3) factors at relapse and duration of response; and (4) prognostic factors in histological transformation. We conclude that, despite the existence of numerous tools, the availability of some of them is still limited; they generally suffer from notable downsides, and most have unproven predictive value, thus having scarce bearing on the choice of regimen at present. However, with the technological and scientific developments of the last few years, the potential for these prognostic factors is promising, particularly in combination, which will probably, in time, help guide therapeutic decisions.
Asunto(s)
Protocolos de Quimioterapia Combinada Antineoplásica , Linfoma Folicular/diagnóstico por imagen , Neoplasia Residual/diagnóstico por imagen , Translocación Genética , Proteína p53 Supresora de Tumor/genética , Médula Ósea/efectos de los fármacos , Médula Ósea/metabolismo , Médula Ósea/patología , Cromosomas Humanos Par 14/química , Cromosomas Humanos Par 18/química , Ensayos Clínicos como Asunto , Supervivencia sin Enfermedad , Humanos , Linfoma Folicular/tratamiento farmacológico , Linfoma Folicular/genética , Linfoma Folicular/mortalidad , Mutación , Clasificación del Tumor , Neoplasia Residual/tratamiento farmacológico , Neoplasia Residual/genética , Neoplasia Residual/mortalidad , Tomografía de Emisión de Positrones , Pronóstico , Recurrencia , Factores de Riesgo , Proteína p53 Supresora de Tumor/metabolismoRESUMEN
Using reverse transcription in conjunction with the quantitative real-time PCR or digital droplet PCR, the transcriptome profiling of human chromosome 18 has been carried out in liver hepatocytes and hepatoblastoma cells (HepG2 cell line) in terms of the absolute number of each transcript per cell. The transcript abundance varies within the range of 0.006 to 9635 and 0.011 to 4819 copies per cell for HepG2 cell line and hepatocytes, respectively. The expression profiles for genes of chromosome 18 in hepatocytes and HepG2 cells were found to significantly correlate: the Spearman's correlation coefficient was equal to 0.81. The distribution of frequency of transcripts over their abundance was bimodal for HepG2 cells and unimodal for liver hepatocytes. Bioinformatic analysis of the differential gene expression has revealed that genes of chromosome 18, overexpressed in HepG2 cells compared to hepatocytes, are associated with cell division and cell adhesion processes. It is assumed that the enhanced expression of those genes in HepG2 cells is related to the proliferation activity of cultured cells. The differences in transcriptome profiles have to be taken into account when modelling liver hepatocytes with cultured HepG2 cells.
Asunto(s)
Cromosomas Humanos Par 18/química , Dosificación de Gen , Hepatocitos/metabolismo , Hígado/metabolismo , ARN Mensajero/genética , Transcriptoma , Biología Computacional , Perfilación de la Expresión Génica , Ontología de Genes , Células Hep G2 , Hepatocitos/citología , Humanos , Hígado/citología , Anotación de Secuencia Molecular , Especificidad de Órganos , Cultivo Primario de Células , ARN Mensajero/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodosAsunto(s)
Ganglios Linfáticos/metabolismo , Linfoma Folicular/genética , Proteínas de Neoplasias/genética , Translocación Genética , Cromosomas Humanos Par 14/química , Cromosomas Humanos Par 18/química , Hibridación Genómica Comparativa , Expresión Génica , Humanos , Ganglios Linfáticos/patología , Linfoma Folicular/diagnóstico , Linfoma Folicular/patología , Proteínas de Neoplasias/metabolismo , Estadificación de Neoplasias , Análisis de Secuencia por Matrices de Oligonucleótidos , PronósticoRESUMEN
Pitt-Hopkins syndrome (PTHS), characterized by severe intellectual disability and typical facial gestalt, is part of the clinical spectrum of Rett-like syndromes. TCF4, encoding a basic helix-loop-helix (bHLH) transcription factor, was identified as the disease-causing gene with de novo molecular defects. While PTHS appears to be a recognizable clinical entity, it seems to remain underdiagnosed, especially when facial gestalt is less typical. With the aim to facilitate the diagnosis of PTHS and to increase its rate and specificity, we have investigated 33 novel patients and defined a Clinical Diagnosis Score. Analysis of 112 individuals (79 previously reported and 33 novel patients) allowed us to delineate the TCF4 mutational spectrum, with 40% point mutations, 30% small deletions/insertions, and 30% deletions. Most of these were private mutations and generated premature stop codons. Missense mutations were localized in the bHLH domain, which is a mutational hotspot. No obvious difference was observed between patients harboring truncating, missense mutations, or deletions, further supporting TCF4 haploinsufficiency as the molecular mechanism underlying PTHS. In this study, we have summarized the current knowledge of TCF4 molecular pathology, reported all the mutations in the TCF4 database (http://www.LOVD.nl/TCF4), and present a novel and comprehensive diagnostic strategy for PTHS.
Asunto(s)
Factores de Transcripción Básicos con Cremalleras de Leucinas y Motivos Hélice-Asa-Hélice/genética , Cromosomas Humanos Par 18/genética , Hiperventilación/diagnóstico , Discapacidad Intelectual/diagnóstico , Factores de Transcripción/genética , Adolescente , Niño , Preescolar , Cromosomas Humanos Par 18/química , Bases de Datos Genéticas , Facies , Femenino , Estudios de Asociación Genética , Variación Genética , Genotipo , Haploinsuficiencia , Haplotipos , Humanos , Hiperventilación/genética , Lactante , Discapacidad Intelectual/genética , Masculino , Mutación Missense , Fenotipo , Estructura Terciaria de Proteína , Eliminación de Secuencia , Inversión de Secuencia , Índice de Severidad de la Enfermedad , Factor de Transcripción 4Asunto(s)
Líquido Amniótico/metabolismo , Biomarcadores/metabolismo , Proteínas Fetales/genética , Regulación del Desarrollo de la Expresión Génica , Pruebas Genéticas/métodos , Polihidramnios/genética , Diagnóstico Prenatal/métodos , Amniocentesis , Líquido Amniótico/química , Antioxidantes/uso terapéutico , Cromosomas Humanos Par 18/química , Cromosomas Humanos Par 18/genética , Cromosomas Humanos Par 21/química , Cromosomas Humanos Par 21/genética , Femenino , Proteínas Fetales/metabolismo , Feto , Expresión Génica , Humanos , Masculino , Hibridación de Ácido Nucleico , Análisis de Secuencia por Matrices de Oligonucleótidos , Polihidramnios/diagnóstico , Polihidramnios/tratamiento farmacológico , Polihidramnios/metabolismo , Polihidramnios/patología , Medicina de Precisión , Embarazo , ARN Mensajero/análisis , ARN Mensajero/biosíntesis , Trisomía/diagnósticoAsunto(s)
Cadenas Pesadas de Inmunoglobulina/metabolismo , Ganglios Linfáticos/metabolismo , Linfoma Folicular , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Translocación Genética/genética , Anciano , Cromosomas Humanos Par 14/química , Cromosomas Humanos Par 14/genética , Cromosomas Humanos Par 18/química , Cromosomas Humanos Par 18/genética , Progresión de la Enfermedad , Evolución Molecular , Expresión Génica , Humanos , Cadenas Pesadas de Inmunoglobulina/genética , Hibridación Fluorescente in Situ , Ganglios Linfáticos/patología , Linfoma Folicular/genética , Linfoma Folicular/patología , Masculino , Estadificación de Neoplasias , Proteínas Proto-Oncogénicas c-bcl-2/genética , Análisis de Secuencia de ADNRESUMEN
Follicular lymphoma (FL) is a germinal centre-derived indolent B-cell lymphoma representing the second most common Non Hodgkin lymphoma in the Western world. This chapter focuses on the pathology of FL and summarizes the current knowledge about genetic and molecular features that are relevant for the pathogenesis of this neoplasm. The translocation t(14;18) is present in approximately 90% of FL leading to the upregulation of the anti-apoptotic protein BCL2, that may constitute a promising molecular target for therapeutic approaches. FL lacking the t(14;18) also exist, and B-cells carrying the t(14;18) can be detected in a subset of healthy individuals. In addition to the t(14;18), secondary genetic alterations are present in most FL and, more recently, deeper insights into the methylation and microRNA expression patterns in the tumour cells have been gained. The tumour microenvironment appears to be particularly important for the biology and the clinical course of FL.
Asunto(s)
Linfocitos B/patología , Cromosomas Humanos Par 14/genética , Cromosomas Humanos Par 18/genética , Centro Germinal/patología , Linfoma de Células B/genética , Linfoma Folicular/genética , Proteínas Proto-Oncogénicas c-bcl-2/genética , Antineoplásicos/uso terapéutico , Linfocitos B/metabolismo , Biomarcadores de Tumor/genética , Biomarcadores de Tumor/metabolismo , Cromosomas Humanos Par 14/química , Cromosomas Humanos Par 18/química , Metilación de ADN , Perfilación de la Expresión Génica , Centro Germinal/metabolismo , Humanos , Linfoma de Células B/tratamiento farmacológico , Linfoma de Células B/metabolismo , Linfoma de Células B/patología , Linfoma Folicular/tratamiento farmacológico , Linfoma Folicular/metabolismo , Linfoma Folicular/patología , MicroARNs/metabolismo , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Translocación Genética , Microambiente TumoralRESUMEN
Follicular lymphoma (FL) grade 3B (FL3B) is defined as FL with more than 15% centroblasts per high resolution field present as solid sheets. Coexistence with diffuse large B-cell lymphoma (DLBCL) is frequent. In contrast to other FL, FL3B frequently lack CD10 expression (approximately 50% of cases), show lower probability of BCL2 expression (69% positive) and increased TP53 expression (31% positive). The t(14;18) hallmark translocation of FL is present in only around 13% of FL3B. In contrast, translocations affecting the BCL6 locus in 3q27 are frequent (44%). Overall, FL3B in many features resembles DLBCL. The presence of a diffuse component in FL3B has been related to an unfavorable outcome except for pediatric FL3B that presents in 60% of the cases this DLBCL component. In this chapter we sought to review the present knowledge on morphological, cytogenetic and molecular features in FL3B.
Asunto(s)
Linfocitos B/patología , Proteínas de Unión al ADN/genética , Linfoma Folicular/genética , Linfoma de Células B Grandes Difuso/genética , Neprilisina/genética , Proteínas Proto-Oncogénicas c-bcl-2/genética , Proteína p53 Supresora de Tumor/genética , Adulto , Linfocitos B/metabolismo , Biomarcadores de Tumor/genética , Biomarcadores de Tumor/metabolismo , Niño , Cromosomas Humanos Par 14/química , Cromosomas Humanos Par 14/genética , Cromosomas Humanos Par 18/química , Cromosomas Humanos Par 18/genética , Comorbilidad , Proteínas de Unión al ADN/metabolismo , Expresión Génica , Humanos , Linfoma Folicular/epidemiología , Linfoma Folicular/metabolismo , Linfoma Folicular/patología , Linfoma de Células B Grandes Difuso/epidemiología , Linfoma de Células B Grandes Difuso/metabolismo , Linfoma de Células B Grandes Difuso/patología , Neprilisina/metabolismo , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Proteínas Proto-Oncogénicas c-bcl-6 , Índice de Severidad de la Enfermedad , Translocación Genética , Proteína p53 Supresora de Tumor/metabolismoRESUMEN
BACKGROUND: This study reports on a hitherto undescribed autosomal recessive syndrome characterised by dysmorphic features and multiple congenital anomalies together with severe neurological impairment, chorea and seizures leading to early death, and the identification of a gene involved in the pathogenesis of the disease. METHODS: Homozygosity mapping was performed using Affymetrix Human Mapping 250k NspI arrays. Sequencing of all coding exons of the candidate genes was performed with primer sets designed using the Primer3 program. Fluorescence activated cell sorting was performed using conjugated antibody to CD59. Staining, acquisition and analysis were performed on a FACSCalibur flow cytometer. RESULTS: Using homozygosity mapping, the study mapped the disease locus to 18q21.32-18q22.1 and identified the disease-causing mutation, c.2126GâA (p.Arg709Gln), in PIGN, which encodes glycosylphosphatidylinositol (GPI) ethanolamine phosphate transferase 1, a protein involved in GPI-anchor biosynthesis. Arginine at the position 709 is a highly evolutionarily conserved residue located in the PigN domain. The expression of GPI linked protein CD59 on fibroblasts from patients as compared to that in a control individual showed a 10-fold reduction in expression, confirming the pathogenic consequences of the mutation on GPI dependent protein expression. CONCLUSIONS: The abundant expression of PIGN in various tissues is compatible with the diverse phenotypic features observed in the patients and with the involvement of multiple body systems. The presence of developmental delay, hypotonia, and epilepsy combined with multiple congenital anomalies, especially anorectal anomalies, should lead a clinician to suspect a GPI deficiency related disorder.
Asunto(s)
Anomalías Múltiples/genética , Antígenos CD59/genética , Trastornos de los Cromosomas/genética , Cromosomas Humanos Par 18/química , Glicosilfosfatidilinositoles/metabolismo , Fosfotransferasas/genética , Transferasas/genética , Anomalías Múltiples/etnología , Árabes/etnología , Secuencia de Bases , Antígenos CD59/metabolismo , Preescolar , Trastornos de los Cromosomas/etnología , Mapeo Cromosómico , Consanguinidad , Exones , Femenino , Citometría de Flujo , Homocigoto , Humanos , Lactante , Israel/epidemiología , Pérdida de Heterocigocidad , Masculino , Datos de Secuencia Molecular , Mutación , Análisis de Secuencia por Matrices de Oligonucleótidos , Linaje , Alineación de Secuencia , SíndromeRESUMEN
The t(X;18)(p11;q11) translocation has been shown to be the specific alteration for synovial sarcomas. The translocation leads to production of chimeric protein SYT/SSX by fusion of SYT and SSX genes involved. The expression analysis of SYT/SSX1 and SYT/SSX2 chimeric transcripts was performed in formalin-fixed soft tissue tumour specimens and the diagnostic validity of immunohistochemistry, FISH and RT-PCR methods was compared. The chimeric transcripts were detected in 12 from 16 synovial sarcomas: 7 SYT/SSX1 and 5 SYT/SSX2 fusion variants; by fluorescence hybridization in situ (FISH) the translocation was found in 13 from 16 sarcoma samples. As synovial sarcoma represents a diagnostically challenging group, genetic analysis of translocations and chimeric transcripts is an extremely useful confirmatory diagnostic tool providing higher sensitivity than immunohistochemistry markers do.
Asunto(s)
Biomarcadores de Tumor/genética , Cromosomas Humanos Par 18/genética , Proteínas de Fusión Oncogénica/genética , Patología Molecular/métodos , Sarcoma Sinovial/genética , Neoplasias de los Tejidos Blandos/genética , Adolescente , Adulto , Anciano , Secuencia de Bases , Biomarcadores de Tumor/análisis , Cromosomas Humanos Par 18/química , Cartilla de ADN/química , Cartilla de ADN/genética , Femenino , Formaldehído , Expresión Génica , Fusión Génica , Humanos , Inmunohistoquímica , Hibridación Fluorescente in Situ , Masculino , Persona de Mediana Edad , Datos de Secuencia Molecular , Proteínas de Fusión Oncogénica/análisis , Adhesión en Parafina , ARN Mensajero/análisis , ARN Mensajero/biosíntesis , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Sarcoma Sinovial/diagnóstico , Sarcoma Sinovial/patología , Neoplasias de los Tejidos Blandos/diagnóstico , Neoplasias de los Tejidos Blandos/patología , Fijación del Tejido , Transcripción Genética , Translocación GenéticaRESUMEN
OBJECTIVE: To establish a multicolor primed in situ labeling (PRINS) protocol for chromosome detection in uncultured amniocytes. METHODS: Chromosomes 18, X and Y in uncultured amniocytes were simultaneously detected by using the non-ddNTP-blocking multicolor PRINS procedure. RESULTS: Within 7 h, the 3 chromosomes were simultaneously marked in the same uncultured amniocyte. The chromosome signals were successfully detected in 69 uncultured samples of amniotic fluid. The results were consistent with that obtained by chromosomes in cultured amniocytes. CONCLUSION: This multicolor protocol was high throughput, fast, simple, sensitive and reliable in diagnosing chromosome abnormalities in uncultured amniocytes.
Asunto(s)
Líquido Amniótico/química , Cromosomas Humanos Par 18/química , Cromosomas Humanos X/química , Cromosomas Humanos Y/química , Etiquetado in Situ Primed/métodos , Adolescente , Adulto , Líquido Amniótico/citología , Células Cultivadas , Cromosomas Humanos Par 18/genética , Cromosomas Humanos X/genética , Cromosomas Humanos Y/genética , Femenino , Humanos , Hibridación Fluorescente in Situ/métodos , Embarazo , Diagnóstico Prenatal/métodos , Adulto JovenRESUMEN
Follicular lymphoma (FL) is characterized by the t(14;18)(q32;q21) chromosomal translocation which can be detected by polymerase chain reaction (PCR) in approximately 70% of cases. The aim of our retrospective study was to evaluate the sensitivity and the reproducibility of both conventional qualitative and quantitative PCR assays for detection of the t(14;18)(q32;q21) chromosomal translocation in biopsy material. Fifty-seven formalin-fixed, paraffin-embedded tumor lymph node (LN) specimens from 50 patients with FL were included in the study. Qualitative PCR was performed with primer sets specific for the MBR, far3'-MBR and the mcr regions, respectively. Quantitative PCR was performed using the LightCycler instrument and the LightCycler - t(14;18) Quantification Kit (MBR). The overall detection rate of the t(14;18) in our study (52.6%) was in accordance with the literature. Of the t(14;18)-positive cases, 49.1% had breakpoints within the MBR and only 3.5% had breakpoints within the mcr. The most sensitive method was LightCycler-based PCR with a detection rate of 47.4%, followed by MBR1,2 assay (43.9%). We observed good agreement between qualitative MBR1,2 and quantitative LightCycler-based assay with a slightly higher detection rate of the quantitative method. The sensitivities of both methods were in accordance with results from other studies. Since LightCycler-based assay detects only breakpoints within the MBR, qualitative PCR should be employed in routine diagnostic settings for detection of breakpoints within the mcr and far3'-MBR regions.
Asunto(s)
Ganglios Linfáticos/patología , Linfoma Folicular/diagnóstico , Linfoma Folicular/patología , Reacción en Cadena de la Polimerasa/métodos , Translocación Genética , Biopsia , Cromosomas Humanos Par 14/química , Cromosomas Humanos Par 18/química , Femenino , Genes de las Cadenas Pesadas de las Inmunoglobulinas , Genes bcl-2 , Humanos , Masculino , Persona de Mediana Edad , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Manejo de EspecímenesRESUMEN
PURPOSE: Existing data suggest that conventional C(H)OP (cyclophosphamide, doxorubicin, vincristine, prednisone) regimen may not be intensive enough to achieve molecular response, as measured by polymerase chain reaction (PCR) evidence of translocation (14;18)(q32:q21) for follicular lymphoma. This study was undertaken to study the molecular response rate of follicular lymphoma to C(H)OP-based therapy and to analyze prognostic factors for molecular response. PATIENTS AND METHODS: Twenty patients with pretreatment PCR evidence of t(14;18)(q32; q21) and at least one posttreatment PCR analysis after the initiation of the treatment with C(H)OP with or without radiation therapy constituted the basis for this analysis. The random effects logistic model was used to analyze the data. The following factors were investigated for their relationship to molecular response: gender, age, beta2-microglobulin, use of radiation therapy, Ann Arbor stage, and international Prognostic Index for malignant lymphoma. RESULTS: Median follow-up was 56 months (range, 23-153 months). A total of 135 PCR results were available, 33 from bone marrow and 102 from peripheral blood. Overall, there was a clear and steady decreasing trend toward loss of PCR positivity with increasing time aftertreatment. By univariate analysis, stage > or = 3, stage = 4, International Prognostic Index > or = 2, and no radiation therapy were adverse factors for molecular response. On multivariate analysis, Ann Arbor stage IV and no radiation therapy were independent risk factors for PCR positivity, both for the peripheral blood data analyzed alone and for all data combined. DISCUSSION: It is possible to achieve molecular response with C(H)OP with or without radiation therapy in patients with follicular lymphoma. Response rate depends on the Ann Arbor stage and radiation therapy.
Asunto(s)
Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Cromosomas Humanos Par 14 , Cromosomas Humanos Par 18 , Ciclofosfamida/uso terapéutico , Doxorrubicina/uso terapéutico , Genes bcl-2/genética , Linfoma Folicular/tratamiento farmacológico , Prednisona/uso terapéutico , Translocación Genética/genética , Vincristina/uso terapéutico , Adulto , Anciano , Cromosomas Humanos Par 14/química , Cromosomas Humanos Par 14/genética , Cromosomas Humanos Par 18/química , Cromosomas Humanos Par 18/genética , Estudios de Evaluación como Asunto , Femenino , Estudios de Seguimiento , Humanos , Modelos Logísticos , Linfoma Folicular/genética , Masculino , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa , Pronóstico , Radioterapia Adyuvante , Resultado del Tratamiento , Microglobulina beta-2/sangreRESUMEN
Myxopapillary ependymomas (MPEs) are low-grade neuroepithelial tumors typically occurring in the conus-cauda equina-filum terminale region. Limited molecular and cytogenetic analysis of MPEs has not demonstrated consistent abnormalities. In an attempt to clarify the chromosomal status of these tumors and identify commonly aberrant regions in the genome we have combined 3 molecular/cyto/genetic methods to study 17 MPEs. Comparative genomic hybridization of 7/17 tumors identified concurrent gain on chromosomes 9 and 18 as the most frequent finding. The majority of the 17 tumors were also studied using microsatellite analysis with marker spanning the whole chromosomes 9 and 18 and interphase-FISH with centromeric probes for both chromosomes. Our combined results were consistent with concurrent gain in both chromosomes 9 and 18 in 11/17 cases, gain of either chromosome 9 or 18 and imbalance in the other chromosome in 3/17 tumors and allelic imbalances of chromosomes 9 or 18 in 3/17 and 1/17 tumors, respectively. Other abnormalities observed included gain of chromosomes 3, 4, 7, 8, 11, 13, 17q, 20, and X and loss of chromosomes 10 and 22. Our findings represent some steps towards understanding the molecular mechanisms involved in the development of MPE.
Asunto(s)
Cromosomas Humanos Par 18/genética , Cromosomas Humanos Par 9/genética , Ependimoma/genética , Neoplasias Neuroepiteliales/genética , Adolescente , Adulto , Anciano , Neoplasias del Sistema Nervioso Central/genética , Neoplasias del Sistema Nervioso Central/patología , Cromosomas Humanos Par 18/química , Cromosomas Humanos Par 9/química , Ependimoma/patología , Femenino , Humanos , Masculino , Repeticiones de Microsatélite/genética , Persona de Mediana Edad , Neoplasias Neuroepiteliales/patología , Neoplasias del Sistema Nervioso Periférico/genética , Neoplasias del Sistema Nervioso Periférico/patologíaRESUMEN
The t(11;18)(q21;q21) between the inhibitor of apoptosis API2 and the MLT gene is a distinct feature of marginal zone B-cell lymphomas of MALT-type. Hitherto the chimeric API2-MLT transcripts are all "in-frame" and predominantly fuse exon 7 of API2 to different MLT exons. Recurrent chromosomal translocations are common in lymphoid neoplasms and might represent by-products of the rearrangement processes generating antigen receptor diversity. The genomic structure of the MLT gene was determined to facilitate amplification of the genomic breakpoint junctions from 5 MALT-type lymphomas with t(11;18). Their sequence analysis showed scattering of the chromosome 11 breakpoints in intron 7 of API2 whereas rearrangements in MLT occurred in intron 2, 4, 7, or 8, respectively. Sequences around the junctions did not display recognition signal sequences mediating lymphocytic V(D)J recombination or other sequence motifs associated with recombination. The breakpoints occurred in a copy of an AluSx repeat in three cases, but interchromosomal Alu-mediated homologous recombination could be ruled out as the repeat resided only on one of the participating chromosomes. The t(11;18) was associated with a deletion in 4 out of 5 cases, ranging in size from 53 bp up to more than 200 kb. These deletions were observed on one or sometimes both derivative chromosomes that might indicate the susceptibility of these regions for breakage. Our data suggest that the API2-MLT fusion might result from a non-homologous end joining event after multiple double-strand breaks. The clustering of breaks in intron 7 of API2 and the consistent "in frame" API2-MLT fusions could therefore reflect certain functional constraints crucial for clonal outgrowth.
Asunto(s)
Rotura Cromosómica/genética , Cromosomas Humanos Par 11/genética , Cromosomas Humanos Par 18/genética , Linfoma de Células B de la Zona Marginal/genética , Proteínas de Neoplasias , Proteínas/química , Proteínas/genética , Translocación Genética/genética , Apoptosis/genética , Secuencia de Bases , Caspasas , Cromosomas Humanos Par 11/química , Cromosomas Humanos Par 18/química , Humanos , Proteínas Inhibidoras de la Apoptosis , Datos de Secuencia Molecular , Proteína 1 de la Translocación del Linfoma del Tejido Linfático Asociado a Mucosas , Proteínas de Fusión Oncogénica/química , Proteínas de Fusión Oncogénica/genética , Análisis de Secuencia de ADN , Neoplasias Gástricas/química , Neoplasias Gástricas/genéticaRESUMEN
Using fluorescence in situ hybridization we show striking differences in nuclear position, chromosome morphology, and interactions with nuclear substructure for human chromosomes 18 and 19. Human chromosome 19 is shown to adopt a more internal position in the nucleus than chromosome 18 and to be more extensively associated with the nuclear matrix. The more peripheral localization of chromosome 18 is established early in the cell cycle and is maintained thereafter. We show that the preferential localization of chromosomes 18 and 19 in the nucleus is reflected in the orientation of translocation chromosomes in the nucleus. Lastly, we show that the inhibition of transcription can have gross, but reversible, effects on chromosome architecture. Our data demonstrate that the distribution of genomic sequences between chromosomes has implications for nuclear structure and we discuss our findings in relation to a model of the human nucleus that is functionally compartmentalized.