Transcriptional Characterization of Glioma Neural Stem Cells Diva ...

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3.3 Glioma Culture Systems Introduction ploidy observed in late passage serum cells. In addition, loss of the wild-type p53 allele in late passage serum cells leaves only the mutant p53 allele found in both the NBE cells and the parental tumours, caus- ing a well-known genomic instability [130,153]. These genomic changes further demonstrate the significant differences between serum cells and their matched parental tumours [261]. In conclusion, by using a model system derived from primary glioblastomas, Lee et al [261] have demonstrated that NBE-cultured cells derived from primary glioblastomas bear remarkable similarity to normal NS cells by retaining the ability to form neurospheres in vitro; an indefinite self-renewal potential; the ability to differentiate into the glial and neuronal lineages of the CNS; hav- ing gene expression profiles similar to NS cells; bearing genetic stability over many passages in vitro; harbour all of the genetic aberrations found within the primary tumour; have gene expression profiles similar to the glioblastomas they were derived from; appear to be the principal tumourigenic cell type that can recapitulate the overall in vivo phenotype of the parental glioblastoma. By contrast, cells derived from the same glioblastoma specimens but grown in serum-containing media lose all the characteristics of primary tumour cultures, although they ultimately regain the tumourigenic potential in later passages without being able to recapitulate the tumourigenic phenotype of the original tumour however, but rather matching the phenotypic and genotypic patterns found in most glioma cell lines. Since several groups have reported that tumour stem cell-like cells can be isolated from the established tumour cell lines by culture in serum-free media with selected growth factors [242,385], experiments need to be carried out to validate how these cells maintain their tumour stem cell-like properties in differentiation-inducing conditions and if cells with stem-like properties may emerge again through epigenetic reprogramming or selection of a subpopula- tion of cells with genomic instability. Based on their findings, Lee et al propose that the inherent tumour stem cell population within primary glioblastomas is quickly lost in typical glioma culture conditions, and the cells found following prolonged in vitro passages are the product of an outgrowth of a cell clone that has undergone profound de novo genetic and/or epigenetic changes. This indicates that NBE cells may be an optimal model system for understanding the biology of primary human tumours, for the preclinical screening of agents and to guide personalized tumour therapy. The table 3.1 below summarises the findings of this study [261]. 76
3.3 Glioma Culture Systems Introduction Table 3.1: Summary of characteristics of NBE and serum-cultured glioblastoma cells. Adapted from Lee et al 2006. Proliferation Constant Clonogenicity, tumourigenicity Differentiation potential Telomerase activity Positive tumour histology Global gene expression NSC-related genes Genotype Glioma Neural Stem Cells NBE-cultured Serum-cultured Limited growth, plateau, exponential growth Yes, regardless of passages Not at early passages Induce to become glial and neuronal lineages Extensive migration, phenocopy of primary human GBMs Similar to primary human GBMs Nestin, Sox2, CD133, Musashi and Bmi Same as parental tumour regardless of passages Do not respond to differentiation stimuli Negative initially, but became positive at late passages Fail to show infiltration like glioma lines Differentiation from primary tumours but similar to common glioma lines - Additional alterations not found in parental tumour The demonstration that the adult human brain maintains areas of radial glia populations that have been shown to give rise to NS cells within the SVZ, raised the prospect that these multipotent cells could be the alternate cells responsible for glioma expansion to the differentiated glia in the brain parenchyma [122,390,399]. As rare populations of stem cells are being discov- ered in different tissues, the cancer stem cell hypothesis reinforces its statement that cell lineage organization in tumours is hierarchical rather than stochastic and only the sub-population of cancer stem cells is responsible for the expansion of the tumour [123,458]. Therefore, in vitro expansion of the putative brain cancer stem cells as stable cell lines would provide a powerful model system to study the human disease by giving insights into the origin of tumour heterogeneity and enable further analysis of the self-renewal, commitment, and differentiation processes, which will hopefully lead to more targeted therapeu- tic strategies [404]. 77
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Transcriptional Characterization of
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This dissertation is my own work an
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Acknowledgements I would like to th
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5.9 External Dataset Expression Cor
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Introduction 1
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1.1 Glial Cells in the Central Nerv
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1.2 Glioblastoma Multiforme Introdu
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1.3 Primary and Secondary Glioblast
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1.4 Pathways Involved in Glioblasto
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6.3 Copy Number Aberrations Results
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6.8 Long ncRNA Differential Express
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Chapter 7 Dataset Correlation Analy
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7.1 Enrichment Analysis Results Tab
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7.1 Enrichment Analysis Results Tab
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8.1 Principles Results say, this is
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8.4 Filters Results Bayesian method
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8.5 Target Prediction Ensemble Anal
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A.1 Differential Expression Appendi
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A.2 Classified Differential Express
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A.3 Quantitative RT-PCR Appendix Ta
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A.3 Quantitative RT-PCR Appendix We
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A.4 Tag-seq vs qRT-PCR Correlation
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Appendix "BEX5", "DIAPH2", "GBP3",
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End Select End If End Sub Appendix
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If NameStr.ToLower().Equals("query"
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Appendix C Long ncRNAs We detected
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C. Long ncRNAs Appendix Tag Accessi
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D.1 Pathway Interactions Appendix F
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D.2 Pathway Images Appendix Figure
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D.2 Pathway Images Appendix Figure
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E. Exon Array Data Appendix Average
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F. MicroRNA Array Data Appendix GNS
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Ln Natural logarithm LOH Loss of He
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7.1 Selected Gene Ontology terms an
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