Project description:Alveolar soft part sarcoma (ASPS) is a soft tissue sarcoma with poor prognosis. We analysed primary and metastatic ASPS samples to elucidate candidate molecular pathways involved in tumor pathogenesis. Differential gene expression analysis revealed the neural origin for ASPS and implication of fusion genes expression in the pathogenesis of ASPS.
Project description:Gene expression from pre- and post- Cediranib treated patients with metastatic Alveolar Soft Part Sarcoma (ASPS) Paired tumor biopsies from 6 patients were collected. Samples were collected at baseline and 3 to 5 days following treatment. The patients are all enrolled in the replicate cohort of a National Cancer Institute (NCI)-sponsered investigational new drug application with institutional review board approval, and all participants provided written informed consent. ClinicalTrials.gov number, NCT00942877. The timing of the tumor biopsies was based on clinical observations of increased tumor pain associated with an inflammatory-like response in peripheral tumor lesions, usually within the second week of treatment. Tumor biopsies were obtained in the first week of treatment before the development of clinical signs and symptoms to evaluate gene expression changes. The time points are labeled "pre" and "post" to denote their relationship to treatment.
Project description:Alveolar soft part sarcoma (ASPS) is a soft tissue sarcoma with poor prognosis. We analysed primary and metastatic ASPS samples to elucidate candidate molecular pathways involved in tumor pathogenesis. Differential gene expression analysis revealed the neural origin for ASPS and implication of fusion genes expression in the pathogenesis of ASPS. Total RNA was extracted, cleaned up, and eluted using the RNeasy MiniKit and RNeasy MinElute Cleanup Kit, respectively (Qiagen), according to the manufacturer’s protocol. The total RNA yield per sample ranged from 191ng to 2ug. cDNA-mediated annealing, selection, ligation, and extension (DASL) expression assay (Illumina) was performed as per manufacturer’s instructions on a panel comprising 24,526 probes for a total of 18,631 genes (Human Ref-8 Expression; BeadChip).
Project description:Alveolar soft-part sarcoma (ASPS) is an extremely rare, highly vascular soft tissue sarcoma affecting predominantly adolescents and young adults. In an attempt to gain insight into the pathobiology of this enigmatic tumor, we performed the first genome-wide gene expression profiling study. Experiment Overall Design: For seven patients with confirmed primary or metastatic ASPS, RNA samples were isolated immediately following surgery, reverse transcribed to cDNA and each sample hybridized to duplicate high-density human U133 plus 2.0 microarrays. Array data was then analyzed relative to arrays hybridized to universal RNA to generate an unbiased transcriptome. Subsequent gene ontology analysis was used to identify transcripts with therapeutic or diagnostic potential. A subset of the most interesting genes was then validated using quantitative RT-PCR and immunohistochemistry.
Project description:Alveolar soft-part sarcoma (ASPS) is an extremely rare, highly vascular soft tissue sarcoma affecting predominantly adolescents and young adults. In an attempt to gain insight into the pathobiology of this enigmatic tumor, we performed the first genome-wide gene expression profiling study.
Project description:Alveolar soft part sarcoma (ASPS) is a soft tissue sarcoma with poor prognosis. Apart from the recurrent, non-reciprocal t(X,17)(p11.2;q25) translocation, there is little molecular evidence for the origin, initiation and progression of this cancer. We analysed 16 primary and metastatic ASPS samples to elucidate candidate molecular pathways involved in tumor pathogenesis. FISH analysis identified the ASPL-TFE3 fusion in all cases. High-resolution aCGH revealed a higher number of numerical aberrations in metastatic tumors relative to primaries, but failed to identify any consistent alterations in either group. Gene expression analysis highlighted 1,063 genes which were differentially expressed between primary and metastatic tumors. Gene set enrichment analysis identified 16 enriched genesets (p < 0.1) associated with differentially expressed genes. Notable among these were several stem cell gene expression signatures and pathways related to differentiation. In particular, the paired box transcription factor PAX6 was up-regulated in the primary tumors, along with several genes whose mouse orthologs have previously been implicated in Pax6-DNA binding during neural stem cell differentiation. In addition to suggesting a neural origin for ASPS, these results implicate transcriptional deregulation from fusion genes in the pathogenesis of ASPS, rather than extensive chromosomal instability. 15 different tumor and normal paired samples were analysed
Project description:Alveolar soft part sarcoma (ASPS) is a rare soft part malignancy affecting adolescents and young adult. ASPS is characterized by its alveolar structure consisting of tumor cells and highly integrated vascular network, and its high metastatic potential indicates the importance of the prominent angiogenic activity of ASPS. Here we find that the expression of ASPSCR1-TFE3, the fusion transcription factor causatively associated with ASPS, is dispensable for in vitro tumor maintenance but required for in vivo tumor development via angiogenesis. ASPSCR1-TFE3 frequently associates with active enhancers including super-enhancers (SE) upon its DNA binding, and the loss of its expression induces dynamic modification of SE distribution related to genes belonging to the angiogenesis pathway. Using epigenomic CRISPR/dCas9 screening, we identify Pdgfb, Rab27a, Sytl2, and Vwf as critical target genes associated with reduced enhancer activities due to the ASPSCR1-TFE3 loss. ASPSCR1-TFE3 thus orchestrates higher ordered angiogenesis via enhanced intracellular trafficking of angiogenic factors.
Project description:Alveolar soft part sarcoma (ASPS) is a rare soft part malignancy affecting adolescents and young adult. ASPS is characterized by its alveolar structure consisting of tumor cells and highly integrated vascular network, and its high metastatic potential indicates the importance of the prominent angiogenic activity of ASPS. Here we find that the expression of ASPSCR1-TFE3, the fusion transcription factor causatively associated with ASPS, is dispensable for in vitro tumor maintenance but required for in vivo tumor development via angiogenesis. ASPSCR1-TFE3 frequently associates with active enhancers including super-enhancers (SE) upon its DNA binding, and the loss of its expression induces dynamic modification of SE distribution related to genes belonging to the angiogenesis pathway. Using epigenomic CRISPR/dCas9 screening, we identify Pdgfb, Rab27a, Sytl2, and Vwf as critical target genes associated with reduced enhancer activities due to the ASPSCR1-TFE3 loss. ASPSCR1-TFE3 thus orchestrates higher ordered angiogenesis via enhanced intracellular trafficking of angiogenic factors.
Project description:In order to dtermine how well a mouse genetic model of alveolar soft part sarcoma (ASPS) mimics the human disease, five human ASPS tumor samples and three normal skeletal muscle samples were profiled by RNAseq and compared to samples from five mouse tumors induced by expression of ASPSCR1-TFE3 and three normal mouse skeletal muscle samples, also profiled by RNAseq. The reference was really comparing 5 human ASPS tumors to 5 mouse tumors that histologically mimic ASPS, but using skeletal muscle controls (3 from each species) as a sounding board for differential expression.