FFPE-samples from cavitational ultrasonic surgical aspirates are suitable for RNA profiling of gliomas
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ABSTRACT: During surgical procedures of gliomas, tissular material obtained from cavitational ultrasonic surgical aspirators (CUSAs) is generally discarded but can exceed amount and quality of certain tumour core resections (TCRs). From archived FFPE-samples, we compared the profiles of somatic mutations and gene expression in the solid material from aspirates and TCRs obtained from the same patients in selected gliomas encompassing grade II to IV. Despite the characteristic heterogeneity of gliomas, we found substantial similarities between the corresponding aspirates and TCRs that included mutations used in the clinical routine and glioma-associated transcriptional signatures. Based on these results, we confirmed the suitability of CUSA-fixed biomaterial from glioma surgeries for downstream applications.
Project description:Glioblastoma stem cells (GSCs) are thought to be the source of tumor growth and therapy resistance. Core biosies that are used to generate GSC cultures are ususally taken from one part of the tumor and are thus unlikely to represent intra-tumoral heterogeneity. This study shows that the ultrasonic aspirates (UA), that are usually considered as a biological waste, can be used as a reliable source of GSCs. Furthermore the UA aspirates seem to be capturing the tumorigenic signature better than the traditional biopsies.
Project description:Diffuse infiltrating gliomas are the most common primary brain malignancy found in adults, and Glioblastoma multiforme, the highest grade glioma, is associated with a median survival of 7 months. Transcriptional profiling has been applied to 85 gliomas from 74 patients to elucidate glioma biology, prognosticate survival, and define tumor sub-classes. These studies reveal that transcriptional profiling of gliomas is more accurate at predicting survival than traditional pathologic grading, and that gliomas characteristically express coordinately regulated genes of one of four molecular signatures: neurogenesis, synaptic transmission, mitotic, or extra-cellular matrix. Elucidation of these survival associated molecular signatures will aid in tumor prognostication and define targets for future directed therapy. Evaluate a large number of diffuse infiltrating gliomas through transcriptional profiling. Glioma tumor sub-classes may be identified through large scale gene expression studies. All patients undergoing surgical treatment at the University of California, Los Angeles for primary brain cancers between 1996 and 2003 were invited to participate in this Institutional Review Board approved study. 74 of the patients participating in this broad protocol were analyzed as part of this study if their initial tumor was diagnosed as a grade III (n=24) or IV (n=50) glioma of any histologic type on initial surgical treatment and fresh frozen material was obtained. Only grade III and IV gliomas were included in this study as the distinction between these grades is subtle and prone to misclassification. The time in days elapsed from resection to the day of death, or if the patient has remained alive, to the current day was recorded for all samples studied. Patient ages at diagnosis varied from 18 to 82 years. There were 46 females and 28 males. Probes were prepared using standard Affymetrix protocols, and hybridized to Affymetrix HG-U133A and HG-U133B arrays.
Project description:Interventions: Experiment Group A:use the ultrasonic surgical systems (Ezisurg Medical Co., Ltd.) in the colorectal surgery;Experiment Group B:use the reusable shear for ultrasonic surgical system (Ezisurg Medical Co., Ltd.) in the colorectal surgery;Control Group:use the Harmonic Ace+7, 5mm Diameter Shears with Advanced Hemostasis (Ethicon Endo-Surgery,LLC) in the colorectal surgery
Primary outcome(s): overall good performance rate
Study Design: Parallel
Project description:Glioblastoma, WHO-grade IV glioma, carries a dismal prognosis owing to its infiltrative growth rates and limited treatment options. Glioblastoma-derived extracellular vesicles (EVs; 30-1000nm membranous particles) influence the microenvironment to mediate tumour aggressiveness and carry oncogenic cargo across the blood-brain-barrier into the circulation. As such, EVs are biomarker reservoirs with enormous potential for assessing glioblastoma tumours in situ. Neurosurgical aspirates are rich sources of EVs, isolated directly from glioma microenvironments. Quantitative LC-MS/MS compared EV proteomes enriched from glioblastoma (n=15) and glioma grade II-III (n=7) aspirates and identified 298 differentially-abundant proteins (p-value<0.00496). Along with previously reported glioblastoma-associated biomarkers, levels of all eight subunits of the key molecular chaperone, T-complex protein 1 Ring complex (TRiC), were higher in glioblastoma-EVs, including CCT2, CCT3, CCT5, CCT6A, CCT7 and TCP1 (p<0.00496). Analogous increases in TRiC transcript levels and DNA copy numbers were detected in silico; CCT6A had the greatest induction of expression and amplification in glioblastoma and showed a negative association with survival (p=0.006). CCT6A is co-localised with EGFR at 7p11.2, with a strong tendency for co-amplification (p<0.001). Immunohistochemistry corroborated the CCT6A proteomics measurements and indicated a potential link between EGFR and CCT6A tissue expressions. Putative EV-biomarkers described here should be further assessed in peripheral blood.
Project description:This study aimed to evaluate the clinical value of copy number variations (CNVs) in fetuses with ultrasonic soft markers. Among 1131 fetuses, 729 had single ultrasonic soft marker, 322 had two ultrasonic soft markers, and 80 had three or more ultrasonic soft markers. All fetuses underwent single nucleotide polymorphism (SNP) array analysis. Among 1131 fetuses with ultrasonic soft markers, 46 had chromosomal abnormalities. In addition to the 46 fetuses with chromosomal abnormalities consistent with the results of the karyotyping analysis, the SNP array identified additional 6.1% (69/1131) abnormal CNVs. No significant difference was found in the rate of abnormal CNVs among the groups. The SNP array can fully complement conventional karyotyping in fetuses with ultrasonic soft markers, improve detection rate of chromosomal abnormalities, and affect pregnancy outcomes.
Project description:Diffuse infiltrating gliomas are the most common primary brain malignancy found in adults, and Glioblastoma multiforme, the highest grade glioma, is associated with a median survival of 7 months. Transcriptional profiling has been applied to 85 gliomas from 74 patients to elucidate glioma biology, prognosticate survival, and define tumor sub-classes. These studies reveal that transcriptional profiling of gliomas is more accurate at predicting survival than traditional pathologic grading, and that gliomas characteristically express coordinately regulated genes of one of four molecular signatures: neurogenesis, synaptic transmission, mitotic, or extra-cellular matrix. Elucidation of these survival associated molecular signatures will aid in tumor prognostication and define targets for future directed therapy. Evaluate a large number of diffuse infiltrating gliomas through transcriptional profiling. Glioma tumor sub-classes may be identified through large scale gene expression studies. All patients undergoing surgical treatment at the University of California, Los Angeles for primary brain cancers between 1996 and 2003 were invited to participate in this Institutional Review Board approved study. 74 of the patients participating in this broad protocol were analyzed as part of this study if their initial tumor was diagnosed as a grade III (n=24) or IV (n=50) glioma of any histologic type on initial surgical treatment and fresh frozen material was obtained. Only grade III and IV gliomas were included in this study as the distinction between these grades is subtle and prone to misclassification. The time in days elapsed from resection to the day of death, or if the patient has remained alive, to the current day was recorded for all samples studied. Patient ages at diagnosis varied from 18 to 82 years. There were 46 females and 28 males. Probes were prepared using standard Affymetrix protocols, and hybridized to Affymetrix HG-U133A and HG-U133B arrays. Keywords: disease state analysis
Project description:We detected fusion genes in 274 fresh surgical samples of gliomas using whole transcriptome sequencing. Using this approach we screened a panel of glioma samples and identified a number of activating novel fusion transcripts. Fusion detection in 274 glioma patients
Project description:Gliomas are aggressive lethal solid brain tumors arising from support cells in the central nervous system. Despite intense efforts to optimize the treatment of gliomas, the outcomes of high grade glioma patients are still frustrating. The causes and progress of gliomas have been investigated extensively; however, the genetic factors involved in the development of this disease remain poorly understood. We used microarrays to detail the global program of gene expression in different grade glioma tissues and try to find out some genes associated with the tumorigenesis of gliomas. Grade I to grade IV glioma tissues were selected in surgical operations for RNA extraction and hybridization on Affymetrix microarrays. We sought to obtain gene expression profiles. To that end, we hand-selected several genes that were differentially expressed in different grade glioma tissues and then performed a further study to identify the role of these genes in the process of the development of gliomas.
Project description:In this study we have performed expression analysis using paired FF-FFPE glioma samples. We show that expression data from FFPE glioma material is concordant with expression data from matched FF tissue, and can be used for molecular profiling in gliomas.