RESUMO
Retinoblastoma (RB, MIM 180200) is the paradigm of hereditary cancer. Individuals harboring a constitutional mutation in one allele of the RB1 gene have a high predisposition to develop RB. Here, we present the first case of familial RB caused by a de novo insertion of a full-length long interspersed element-1 (LINE-1) into intron 14 of the RB1 gene that caused a highly heterogeneous splicing pattern of RB1 mRNA. LINE-1 insertion was inferred by mRNA studies and full-length sequenced by massive parallel sequencing. Some of the aberrant mRNAs were produced by noncanonical acceptor splice sites, a new finding that up to date has not been described to occur upon LINE-1 retrotransposition. Our results clearly show that RNA-based strategies have the potential to detect disease-causing transposon insertions. It also confirms that the incorporation of new genetic approaches, such as massive parallel sequencing, contributes to characterize at the sequence level these unique and exceptional genetic alterations.
Assuntos
Processamento Alternativo , Íntrons , Elementos Nucleotídeos Longos e Dispersos , Mutagênese Insercional , Proteína do Retinoblastoma/genética , Retinoblastoma/genética , Adulto , Biologia Computacional/métodos , Éxons , Loci Gênicos , Humanos , Lactente , Masculino , Retinoblastoma/diagnóstico , Análise de Sequência de DNARESUMO
Medullary thyroid carcinoma accounts for 2% to 5% of thyroid malignancies, of which 75% are sporadic and the remaining 25% are hereditary and related to multiple endocrine neoplasia type 2 syndrome. Despite a genotype-phenotype correlation with specific germline RET mutations, knowledge of pathways specifically associated with each mutation and with non-RET-mutated sporadic MTC remains lacking. Gene expression patterns have provided a tool for identifying molecular events related to specific tumor types and to different clinical features that could help identify novel therapeutic targets. Using transcriptional profiling of 49 frozen MTC specimens classified as RET mutation, we identified PROM1, LOXL2, GFRA1, and DKK4 as related to RET(M918T) and GAL as related to RET(634) mutation. An independent series of 19 frozen and 23 formalin-fixed, paraffin-embedded (FFPE) MTCs was used for validation by RT-qPCR. Two tissue microarrays containing 69 MTCs were available for IHC assays. According to pathway enrichment analysis and gene ontology biological processes, genes associated with the MTC(M918T) group were involved mainly in proliferative, cell adhesion, and general malignant metastatic effects and with Wnt, Notch, NFκB, JAK/Stat, and MAPK signaling pathways. Assays based on silencing of PROM1 by siRNAs performed in the MZ-CRC-1 cell line, harboring RET(M918T), caused an increase in apoptotic nuclei, suggesting that PROM1 is necessary for survival of these cells. This is the first report of PROM1 overexpression among primary tumors.
Assuntos
Biomarcadores Tumorais/genética , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Predisposição Genética para Doença , Neoplasias da Glândula Tireoide/genética , Antígeno AC133 , Antígenos CD/metabolismo , Apoptose/genética , Carcinoma Neuroendócrino , Linhagem Celular Tumoral , Análise por Conglomerados , Técnicas de Silenciamento de Genes , Inativação Gênica , Glicoproteínas/metabolismo , Humanos , Imuno-Histoquímica , Padrões de Herança/genética , Peptídeos/metabolismo , RNA Interferente Pequeno/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Neoplasias da Glândula Tireoide/patologiaRESUMO
The six major genes involved in hereditary susceptibility for pheochromocytoma (PCC)/paraganglioma (PGL) (RET, VHL, NF1, SDHB, SDHC, and SDHD) have been recently integrated into the same neuronal apoptotic pathway where mutations in any of these genes lead to cell death. In this model, prolyl hydroxylase 3 (EglN3) abrogation plays a pivotal role, but the molecular mechanisms underlying its inactivation are currently unknown. The aim of the study was to decipher specific alterations associated with the different genetic classes of PCCs/PGLs. With this purpose, 84 genetically characterized tumors were analyzed by means of transcriptional profiling. The analysis revealed a hypoxia-inducible factor (HIF)-related signature common to succinate dehydrogenase (SDH) and von Hippel-Lindau (VHL) tumors, that differentiated them from RET and neurofibromatosis type 1 cases. Both canonical HIF-1α and HIF-2α target genes were overexpressed in the SDH/VHL cluster, suggesting that a global HIF deregulation accounts for this common profile. Nevertheless, when we compared VHL tumors with SDHB cases, which often exhibit a malignant behavior, we found that HIF-1α target genes showed a predominant activation in the VHL PCCs. Expression data from 67 HIF target genes was sufficient to cluster SDHB and VHL tumors into two different groups, demonstrating different pseudo-hypoxic signatures. In addition, VHL-mutated tumors showed an unexpected overexpression of EglN3 mRNA that did not lead to significantly different EglN3 protein levels. These findings pave the way for more specific therapeutic approaches for malignant PCCs/PGLs management based on the patient's genetic alteration.
Assuntos
Morte Celular/genética , Paraganglioma/genética , Feocromocitoma/genética , Succinato Desidrogenase/genética , Proteína Supressora de Tumor Von Hippel-Lindau/genética , Adolescente , Neoplasias das Glândulas Suprarrenais/genética , Neoplasias das Glândulas Suprarrenais/metabolismo , Adulto , Idoso , Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Fatores de Transcrição Hélice-Alça-Hélice Básicos/metabolismo , Criança , Dioxigenases/genética , Dioxigenases/metabolismo , Humanos , Subunidade alfa do Fator 1 Induzível por Hipóxia/genética , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Prolina Dioxigenases do Fator Induzível por Hipóxia , Pessoa de Meia-Idade , Neoplasias/genética , Paraganglioma/metabolismo , Feocromocitoma/metabolismo , Succinato Desidrogenase/metabolismo , Proteína Supressora de Tumor Von Hippel-Lindau/metabolismo , Adulto Jovem , Doença de von Hippel-Lindau/genética , Doença de von Hippel-Lindau/metabolismoRESUMO
El desafío actual de la Administración Pública y especialmente del Ministerio de Salud Pública de la Rioja, a través de la Dirección General de Administración debe estar orientado al utilización de un modelo, que contemplando los cambios tecnológicos (revolución tecnológica basada en la información), cambios en el ciudadano ( necesidades que evolucionan y ciudadanos más exigentes) y cambios en el entorno, entornos complejos, muchos agentes implicados necesidades de innovación que trasformen el modelo de gestión tradicional, jerárquico, burocrático, de gestión por normas, con concentración de decisiones, manual, por un sistema de gestión participativo, de liderazgo compartido, de gestión proactiva, de decisiones compartidas, de búsqueda de la eficiencia y eficacia y fundamentalmente de gestión humana compleja, integridad (compromiso), empoderamiento e iniciativa...
The current challenge of the Public Administration and especially of the Public Health Ministry of La Rioja, within the general office of Administracion, must be oriented to the use of a model that contmplates the technological changes (technological revolution based on information), changes within citizens (evolving needs and more demandig citizens) and changes in the administrative environment. The complex administrative environment involves many agents and needs of innovation to transform the traditional, hierarchical, and bureaucratic model of administration, which concentrates in manual decisions, into an administrative and participative system of shared leadership, proactive management that seeks for efficacy and efficiency and above all integrity (commitment), empowerment and initiative...