Assuntos
Adenocarcinoma/terapia , Biomarcadores Tumorais/genética , Neoplasias da Vesícula Biliar/terapia , Mesotelioma/terapia , Planejamento de Assistência ao Paciente , Neoplasias Peritoneais/terapia , Medicina de Precisão , Adenocarcinoma/diagnóstico , Adenocarcinoma/genética , Diagnóstico Diferencial , Neoplasias da Vesícula Biliar/diagnóstico , Neoplasias da Vesícula Biliar/genética , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Masculino , Mesotelioma/diagnóstico , Mesotelioma/genética , Pessoa de Meia-Idade , Terapia de Alvo Molecular , Mutação , Neoplasias Peritoneais/diagnóstico , Neoplasias Peritoneais/genética , PrognósticoRESUMO
Primary mediastinal (thymic) large B-cell lymphoma (PMBL) is described as almost always negative for Epstein-Barr virus (EBV). In the context of a mediastinal lymphoma, the distinction between PMBL, classical Hodgkin lymphoma, diffuse large B-cell lymphoma, and mediastinal gray-zone lymphoma can be very difficult; hence, EBV positivity often argues against PMBL. We present a 19-year-old man with mediastinal mass morphologically consistent with PMBL. The tumor expressed classic immunophenotype, including positivity for CD20, CD19, MAL, OCT2, BOB1, BCL6, CD79a, and subset positivity for CD30. However, the tumor was EBV-positive by in situ hybridization. Next-generation sequencing detected somatic mutations in XPO1 (E571K), SMARCB1 (L356fs), and MYCC (T73A). Although the immunophenotype and XPO1 mutation are characteristic of PMBL, EBV expression is uncommon. Since EBV positivity can occur in rare PMBLs, it should not be the deciding factor in the diagnosis. This is the first EBV-positive PMBL in which mutational profiling has been reported. Aside from providing diagnostic support, the finding of the XPO1 E571K mutation may suggest a targeted therapeutic option.
Assuntos
Biomarcadores Tumorais/metabolismo , Infecções por Vírus Epstein-Barr/diagnóstico , Herpesvirus Humano 4/fisiologia , Carioferinas/metabolismo , Linfoma Difuso de Grandes Células B/diagnóstico , Neoplasias do Mediastino/diagnóstico , Receptores Citoplasmáticos e Nucleares/metabolismo , Neoplasias do Timo/diagnóstico , Adulto , Diagnóstico Diferencial , Infecções por Vírus Epstein-Barr/genética , Infecções por Vírus Epstein-Barr/patologia , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Imuno-Histoquímica , Imunofenotipagem , Carioferinas/genética , Carioferinas/imunologia , Linfoma Difuso de Grandes Células B/genética , Linfoma Difuso de Grandes Células B/patologia , Masculino , Mutação/genética , Receptores Citoplasmáticos e Nucleares/genética , Receptores Citoplasmáticos e Nucleares/imunologia , Neoplasias do Timo/genética , Neoplasias do Timo/patologia , Adulto Jovem , Proteína Exportina 1Assuntos
Comitês Consultivos/organização & administração , Institutos de Câncer/organização & administração , Prestação Integrada de Cuidados de Saúde/organização & administração , Oncologia/organização & administração , Equipe de Assistência ao Paciente/organização & administração , Medicina de Precisão , Connecticut , Comportamento Cooperativo , Humanos , Comunicação InterdisciplinarAssuntos
Terapia de Alvo Molecular , Ftalazinas/uso terapêutico , Piperazinas/uso terapêutico , Inibidores de Poli(ADP-Ribose) Polimerases/uso terapêutico , Neoplasias da Próstata/tratamento farmacológico , Sarcoma/tratamento farmacológico , Idoso , Proteínas Mutadas de Ataxia Telangiectasia/genética , Biomarcadores Tumorais/genética , Progressão da Doença , Evolução Fatal , Humanos , Masculino , Pessoa de Meia-Idade , Mutação , Medicina de Precisão , Neoplasias da Próstata/genética , Neoplasias da Próstata/secundário , Sarcoma/genética , Sarcoma/secundário , Resultado do TratamentoRESUMO
The National Cancer Institute-Molecular Analysis for Therapy Choice (NCI-MATCH) trial is a national signal-finding precision medicine study that relies on genomic assays to screen and enroll patients with relapsed or refractory cancer after standard treatments. We report the analytical validation processes for the next-generation sequencing (NGS) assay that was tailored for regulatory compliant use in the trial. The Oncomine Cancer Panel assay and the Personal Genome Machine were used in four networked laboratories accredited for the Clinical Laboratory Improvement Amendments. Using formalin-fixed paraffin-embedded clinical specimens and cell lines, we found that the assay achieved overall sensitivity of 96.98% for 265 known mutations and 99.99% specificity. High reproducibility in detecting all reportable variants was observed, with a 99.99% mean interoperator pairwise concordance across the four laboratories. The limit of detection for each variant type was 2.8% for single-nucleotide variants, 10.5% for insertion/deletions, 6.8% for large insertion/deletions (gap ≥4 bp), and four copies for gene amplification. The assay system from biopsy collection through reporting was tested and found to be fully fit for purpose. Our results indicate that the NCI-MATCH NGS assay met the criteria for the intended clinical use and that high reproducibility of a complex NGS assay is achievable across multiple clinical laboratories. Our validation approaches can serve as a template for development and validation of other NGS assays for precision medicine.
Assuntos
Sequenciamento de Nucleotídeos em Larga Escala , Neoplasias/diagnóstico , Neoplasias/genética , Ensaios Clínicos como Assunto , Biologia Computacional/métodos , Variação Genética , Genômica/métodos , Genômica/normas , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Sequenciamento de Nucleotídeos em Larga Escala/normas , Humanos , Garantia da Qualidade dos Cuidados de Saúde , Controle de Qualidade , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Fluxo de TrabalhoRESUMO
The Polycomb repressive complex 2 (PRC2), which contains three core proteins EZH2, EED and SUZ12, controls chromatin compaction and transcription repression through trimethylation of lysine 27 on histone 3. The (7;17)(p15;q21) chromosomal translocation present in most cases of endometrial stromal sarcomas (ESSs) results in the in-frame fusion of the JAZF1 and SUZ12 genes. We have investigated whether and how the fusion protein JAZF1-SUZ12 functionally alters PRC2. We found that the fusion protein exists at high levels in ESS containing the t(7;17). Co-transient transfection assay indicated JAZF1-SUZ12 destabilized PRC2 components EZH2 and EED, resulting in decreased histone methyl transferase (HMT) activity, which was confirmed by in vitro studies using reconstituted PRC2 and nucleosome array substrates. We also demonstrated the PRC2 containing the fusion protein decreased the binding affinity to target chromatin loci. In addition, we found that trimethylation of H3K27 was decreased in ESS samples with the t(7;17), but there was no detectable change in H3K9 in these tissues. Moreover, re-expression of SUZ12 in Suz12 (-/-) ES cells rescued the neuronal differentiation while the fusion protein failed to restore this function and enhanced cell proliferation. In summary, our studies reveal that JAZF1-SUZ12 fusion protein disrupts the PRC2 complex, abolishes HMT activity and subsequently activates chromatin/genes normally repressed by PRC2. Such dyesfunction of PRC2 inhibits normal neural differentiation of ES cell and increases cell proliferation. Related changes induced by the JAZF-SUZ12 protein in endometrial stromal cells may explain the oncogenic effect of the t(7;17) in ESS.
