RESUMO
Acute leukemias (AL) comprise a heterogeneous group of hematologic malignancies, and individual patient responses to treatment can be difficult to predict. Monitoring of minimal residual disease (MRD) is thus very important and holds great potential for improving treatment strategies. Common MRD targets include recurrent cytogenetic abnormalities and mutations in important hematological genes; unfortunately well-characterized targets are lacking in many AL patients. Here we demonstrate a technical approach for the identification and mapping of novel clone-specific chromosomal abnormalities down to the nucleotide level. We used molecular cytogenetics, chromosome microdissection, amplification of the microdissected material, and next-generation sequencing to develop PCR-based MRD assays based on unique breakpoint sequences.
Assuntos
Biomarcadores Tumorais/genética , Leucemia/diagnóstico , Leucemia/genética , Neoplasia Residual/diagnóstico , Neoplasia Residual/genética , Cariótipo Anormal , Sequência de Bases , Linhagem Celular Tumoral , Aberrações Cromossômicas , Bandeamento Cromossômico , Pontos de Quebra do Cromossomo , Mapeamento Cromossômico , Proteínas de Ligação a DNA/genética , Histona-Lisina N-Metiltransferase , Humanos , Células K562 , Proteína de Leucina Linfoide-Mieloide/genética , Proteínas Nucleares/genética , Proteínas de Fusão Oncogênica/genética , Fatores de Elongação da TranscriçãoRESUMO
Hematooncologic patients often host rare or fastidious pathogens. Using 16S rDNA sequencing and transmission electron microscopy, we have identified 2 lymphoma patients infected with Candidatus Neoehrlichia mikurensis. In both individuals, the clinical presentation suggested ehrlichiosis-like syndrome. We believe that molecular techniques open new vistas in the field of pathogen detection.
Assuntos
Infecções por Anaplasmataceae/diagnóstico , Anaplasmataceae/classificação , Anaplasmataceae/isolamento & purificação , Técnicas de Diagnóstico Molecular , Anaplasmataceae/genética , Infecções por Anaplasmataceae/complicações , Animais , DNA Bacteriano/genética , DNA Ribossômico/genética , Ehrlichiose/diagnóstico , Feminino , Febre de Causa Desconhecida/diagnóstico , Febre de Causa Desconhecida/microbiologia , Neoplasias Hematológicas/complicações , Humanos , Hospedeiro Imunocomprometido , Masculino , Microscopia Eletrônica , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , Análise de Sequência de DNA , Carrapatos/microbiologiaRESUMO
TP53 plays a pivotal role in the process of DNA repair and apoptosis. In 10-20% of patients with chronic lymphocytic leukemia (CLL), the TP53 pathway is affected. In this study, we analyzed the TP53 mutation status in 2435 consecutive CLL samples, including 1287 diagnostic samples and 1148 samples during follow-up, using FASAY (Functional Analysis of Separated Alleles in Yeast) and direct sequencing. In a cohort of 1287 diagnostic CLL samples, we identified 237 cases with TP53 variants, including mutations, temperature-sensitive variants, deletions, insertions and aberrant splicing variants (18.4%). In 1148 follow-up samples, no TP53 clonal evolution was observed.
