RESUMO
Dystrophia myotonica type 1 (DM1), the most common muscular dystrophy in adults, results from expansion of a CTG repeat in the 3'-untranslated region of the dystrophia myotonica protein kinase gene (DMPK). Correction of the mutant DMPK transcript is a potential therapeutic strategy in DM1. We investigated the efficacy of artificial trans-splicing molecules (ATMs) to target and correct DMPK transcripts. ATMs designed to target intron 14 of DMPK pre-mRNA transcripts were tested for their ability to trans-splice the transcripts of a DMPK mini-gene construct and the endogenous DMPK transcripts of human myosarcoma cells (CCL-136). On agarose gel electrophoresis analysis, six of eight ATMs showed trans-splicing efficacy when applied to DMPK mini-gene construct transcripts, of which three were able to trans-splice endogenous DMPK pre-mRNA transcripts in myosarcoma cells, with trans-splicing efficiency ranging from 1.81 to 7.41%. These findings confirm that artificial trans-splicing can repair DMPK pre-mRNA and provide proof-of-principle evidence for this approach as potential therapeutic strategy for DM1.
Assuntos
Distrofia Miotônica/genética , Proteínas Serina-Treonina Quinases/genética , Precursores de RNA/genética , Trans-Splicing/genética , Sequência de Bases , Marcação de Genes/métodos , Humanos , Dados de Sequência Molecular , Miossarcoma/enzimologia , Miossarcoma/genética , Distrofia Miotônica/enzimologia , Miotonina Proteína Quinase , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Transfecção , Células Tumorais CultivadasRESUMO
Myogenic sarcomas include soft tissue sarcomas that show skeletal muscle differentiation (rhabdomyosarcoma) and those with smooth muscle differentiation (leiomyosarcoma). Rhabdomyosarcomas are more common in the pediatric age group and leiomyosarcomas occur more often in the adult population. Based on the clinico-pathologic features and genetic abnormalities identified, the rhabdomyosarcomas are classified into embryonal, alveolar, spindle cell/sclerosing, and pleomorphic subtypes. Each subtype shows distinctive morphology and has characteristic genetic abnormalities. In this update on myogenic sarcomas, each entity is discussed with special emphasis on recent updates in genetic findings and the diagnostic approach to these tumors.
Assuntos
Miossarcoma/diagnóstico , Miossarcoma/genética , Neoplasias de Tecidos Moles/diagnóstico , Neoplasias de Tecidos Moles/genética , Biomarcadores Tumorais/genética , Diferenciação Celular , Diagnóstico Diferencial , Humanos , Imuno-Histoquímica , Mutação/genética , Proteína MyoD/genética , Miossarcoma/classificação , Miossarcoma/patologia , Prognóstico , Neoplasias de Tecidos Moles/classificação , Neoplasias de Tecidos Moles/patologiaRESUMO
We describe four patients with retroperitoneal liposarcomas undergoing myosarcomatous differentiation. The patients (two men and two women) were 47, 48, 68, and 72 years of age when first seen. The primary tumors were large retroperitoneal, well-differentiated liposarcomas, one featuring areas of dedifferentiation (without muscle elements). A myosarcomatous component became evident at the first recurrence in three cases and at the second recurrence in one. This component was always within dedifferentiated areas and in three of the cases coincided with the emergence of the latter. The muscle component had exclusively leiomyosarcomatous phenotype (alpha-smooth-muscle actin reactivity) in one case, exclusively rhabdomyosarcomatous phenotype (myoglobin reactivity) in two cases, and combined leiomyosarcomatous and rhabdomyosarcomatous phenotype (alpha-smooth-muscle actin and myoglobin) in one case. Ultrastructural studies of one of the tumors with a rhabdomyosarcomatous component revealed the presence of sarcomeres. Two patients died of extensive retroperitoneal disease, one patient died following the attempted removal of a recurrence, and one patient is alive and free of disease. These cases demonstrate that the dedifferentiated component of liposarcoma may exhibit a myosarcomatous component, a feature analogous to that previously described in dedifferentiated chondrosarcoma.
Assuntos
Lipossarcoma/patologia , Miossarcoma/patologia , Neoplasias Retroperitoneais/patologia , Idoso , Feminino , Humanos , Lipossarcoma/genética , Masculino , Pessoa de Meia-Idade , Miossarcoma/genética , Recidiva Local de Neoplasia , Fenótipo , Neoplasias Retroperitoneais/genéticaRESUMO
We investigated the molecular cytogenetic features in myofibroblastic sarcoma (MS) to gain insight into the nature of the controversial entity. DNA copy number changes were analyzed by comparative genomic hybridization in 29 cases of MS and 5 cases of nodular fasciitis. The characteristic chromosomal imbalances in MS were gains at 1p11 --> p36.3 (19/29 [66%]), 12p12.2 --> p13.2 (13/29 [45%]), 5p13.2 --> p15.3 (9/29 [31%]), and chromosome 22 (8/29 [28%]) and loss at 15q25 --> q26.2 (7/29 [24%]). In contrast, only 1 of 5 cases of nodular fasciitis showed genetic aberrations. The average number of aberrations in nodular fasciitis (0.4) was significantly lower than that in MS (5.4). Thus, MS displayed complex DNA copy number changes and shared no range of common chromosomal abnormality with nodular fasciitis, indicating that distinct genetic pathways may be involved in the development of these entities.
Assuntos
Aneuploidia , Fasciite/genética , Miossarcoma/genética , Neoplasias de Tecidos Moles/genética , Adolescente , Adulto , Idoso , Hibridização Genômica Comparativa , DNA de Neoplasias/análise , Fasciite/patologia , Feminino , Dosagem de Genes , Humanos , Cariotipagem , Masculino , Pessoa de Meia-Idade , Miossarcoma/secundário , Recidiva Local de Neoplasia , Neoplasias de Tecidos Moles/patologia , Adulto JovemRESUMO
With the aim of identifying objective cytogenetic-morphologic correlations, we evaluated 46 pleomorphic soft tissue sarcomas (mainly diagnosed originally as malignant fibrous histiocytomas) with clonal chromosome aberrations both cytogenetically and morphologically as part of an international collaborative study. By detailed histopathologic examination, most cases could be categorized into specific tumor types. Eight sarcomas were diagnosed as lipogenic (4 pleomorphic, 1 combined pleomorphic and myxoid/round cell, and 3 dedifferentiated liposarcomas), 19 as myogenic [11 leiomyosarcomas, 1 rhabdomyosarcoma, 4 myosarcomas not otherwise specified (NOS), and 3 probable myosarcomas NOS], 8 as myxofibrosarcomas, 1 as a malignant peripheral nerve sheath tumor, 1 as malignant mesenchymoma, 1 as extraskeletal osteosarcoma, I as sarcoma resembling proliferative fasciitis, and 7 as pleomorphic sarcomas NOS. In a three-grade system, 10 tumors were grade 2 and 36 were grade 3. The majority had highly complex karyotypes. A total of 24 recurrent abnormalities (defined by their presence in at least five cases) were detected: ring chromosomes, homogeneously staining regions (hsr) and/or double minute chromosomes (dmin), and structural rearrangement of 22 different chromosome bands or regions. The frequency and distribution of the recurrent karyotypic features were uneven. Grade 3 tumors displayed, on average, more aberrations per case than did grade 2 tumors. Nine of the selected abnormalities, including hsr/dmin and rearrangements of 19p13 and 19q13, were found only among the high-grade tumors. When the tumors were subdivided according to lineage of differentiation, the highest frequency of aberrations was seen in pleomorphic sarcomas NOS, followed by myxofibrosarcomas, myogenic sarcomas, and lipogenic sarcomas. None of the selected rearrangements was, however, specific for any of these subgroups. The sole consistent cytogenetic-morphologic association was that all three dedifferentiated liposarcomas had multiple abnormal clones, at least one of which contained supernumerary ring chromosomes. Due mainly to karyotype complexity, it therefore seems unlikely that cytogenetic analysis can assist in the differential diagnostic subclassification of pleomorphic sarcomas, nor was there any clear-cut indication that the karyotypic picture could be used to predict clinical outcome. Although the mean number of recurrent chromosome aberrations was almost twice as high in sarcomas that gave rise to metastases as among those that did not, no particular aberration was restricted to either of the two subgroups.