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Studies of CD38 in chronic lymphocytic leukaemiaMatrai, Zoltan January 2004 (has links)
This Thesis consists of 4 experimental Chapters preceded by a General Introduction and followed by a brief section dealing with conclusions and future. In Chapter 1, the prognostic value of CD38 and the relationship between surface expression of this molecule and IgVH mutation are considered. It is shown that surface CD38 expression on CLL cells is highly predictive of nonhypermutated IgVH gene mutation status, while CD38 negativity lacks such predictive value, since IgVH mutated and uninutated cases occur with approximately equal frequency in this subgroup. It is also shown, in agreement with the literature, that CD38 expression is associated with shorter survival and male preponderance. Abundant data are available on the intracellular expression of CD38 in other cell types, but not in CLL. Therefore, Chapter 2 deals with intracellular CD38 in CLL cells and with the different molecular forms expressed in sCD38+ and sCD38- CLL clones. Using a number of different techniques, it is shown that CD38 is expressed intracellularly in all CLL cells irrespectively of the surface expression of the molecule. In keeping with these findings, CD38 mRNA was found in both sCD38+ and sCD38- CLL clones at comparably low levels. It is documented by Western blotting and immunoprecipitation that CLL cells express, in addition to the 45 kD CD38 monomer, other molecular forms of 27, 60 and 205 M Among these, the high molecular weight molecule probably represents a tetrameric form of CD38 and is described in CLL for the first time. A main difference found between the sCD38+ and sCD38- clones was the almost complete absence of the 45 kD monomeric form in the sCD38- clones. An unexpected and interesting finding was a surface immunoreactivity with the anti- CD38 antibody Ab-4 on CLL cells previously classified as IICD38 negative" with the HB-7 antibody. In Chapter 3, the topology of different molecular forms of CD38 is studied further and their enzymatic functions are examined. By subcellular fractionation it is demonstrated that the tetrameric form of CD38 is most abundant in the membrane fraction. Surface radioiodination of CLL cells indicated, for the first time that the main forms of CD38 on the surface of CLL cells are the 45 kD and the -205 kD molecules. The 205 kD molecule was demonstrable on both sCD38+ and sCD38- cells, while the 45 kD molecule was present only on sCD38+ cells. Surface forms of CD38 on CLL cells were characterised further by enzymatic assays performed before and after protease digestion of surface proteins. Results indicated that surface forms are the major sources of CD38 enzymatic activity in CLL cells. Total cyclase and hydrolase activities were also compared in sCD38+ and sCD38- CLL cells and it was shown that both cell types possess these enzymatic functions, although activities were lower in sCD38- cells. It seems likely that the 45 kD molecule (probably present as non-covalently linked dimers) is responsible for the greater enzymatic activity of sCD38+ clones. Enzymatic assays performed after recovering proteins from polyacrylamide gels indicated that eluates from the HMW (>116 kD) section of the gels containing the putative CD38 tetramers have both cyclase and hydrolase activities.Chapter 4 addresses the question of why CD38-positive CLL is characterised by progressiveness and poor outcome. Since CD38 has a welldocumented role in cell cycle regulation and cell proliferation in other cell types, it was hypothesized that a similar linkage might be also present in CLL, contributing to the the adverse clinical outcome. To test this hypothesis, CLL cells were co-stained for surface CD38 and nuclear Ki-67 following a saponinbased permeabilization procedure. CLL clones classified as CD38+ expressed significantly higher levels of Ki-67 than did sCD38- clones. Clones classified as Ki-67+ (>5%) expressed significantly higher levels of CD38 than did Ki-67- ones. Relating surface CD38% and nuclear Ki-67% revealed a linear correlation between these two parameters. Also, the Ki-67+ subpopulation within a given CLL clone expressed significantly higher CD38 values than did the Ki-67- subpopulation. Furthermore, larger CLL cells showed significantly higher Ki-67 values than small CLL lymphocytes. Finally, CD38 immunoreactivity in CLL lymph node sections was found to be strongest in the proliferation centres. These data therefore indicate, for the first time, that surface CD38 expression is a marker of cell cycling activity in CLL and help to explain the adverse prognosis of CD38+ CLL and CLUPL.
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Gene expression analysis of telomerase related genes in myeloid malignancyCampbell, Lisa Jane January 2011 (has links)
Telomere shortening and an increased telomerase activity are associated with poor prognosis and disease progression in many cancers. In Chronic Myeloid Leukaemia (CML) telomere shortening has a strong correlation with disease progression. Expression of hTERT, the catalytic component of telomerase, was evaluated in the CD34+ cells of CML patients. This revealed that expression of hTERT was significantly reduced in chronic phase CML and decreased with disease progression to accelerated phase and blast crisis. .It could therefore be concluded that reduced hTERT expression contributes to reduced telomere length in CML. Additionally, expression of c-Myc, which increases hTERT transcription, correlated with hTERT expression suggesting decreased hTERT is partly caused by reduced c-Myc. hTERT promoter methylation and mutation status were investigated and this revealed that the hTERT promoter was not methylated and mutation rates were low suggesting that these are not contributing to reduced hTERT expression. cDNA microarrays were used to analyse gene expression in neutrophils of patients with Essential Thrombocythaemia (ET) harbouring the JAK2 V617F mutation which, like the BCRlABL translocation in CML, results in an activated kinase. Neutrophils of ET patients exhibited a gene expression profile close to that of controls despite the presence of the mutation. Affymetrix microarrays were used to investigate the role of telomerase related genes in Myelodysplastic Syndromes (MDS). Genes decreased in patients with del(5q) include positive regulators of telomere length and genes with higher expression were associated with increased telomerase activity.
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The role of cytogenetics in leukaemiaSlater, Sarah January 2000 (has links)
No description available.
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A fluorescence in-situ hybridisation and molecular investigation of leukaemia associated abnormalities of chromosome 6qSinclair, Paul Burdwood January 2002 (has links)
Deletions of the long arm of chromosome 6 are a recognised recurrent cytogenetic abnormality associated with ALL. Balanced rearrangements of 6q are less common but occur in leukaemias of both lymphoid and myeloid origin. With the aim of identifying genes that contribute to leukaemia through loss or rearrangement the breakpoints of translocations and deletions of 6q were mapped by FISH. Comparison between the mapped deletions led to the identification of a 4.8 Mb CDR and candidate tumour suppressor gene (GluR-6). Expression of GluR-6 was demonstrated to occur in haernatologic tissues and mutation analysis was performed on leukaemic samples with clonal deletions of the region. In one of 14 cases a base pair substitution that led to a change in amino acid sequence was found. The base pair substitution was also present in the patients' remission sample but was not seen in any of 20 other normal bone marrow samples analysed. Analysis of the translocations identified a variety of breakpoints between 6q15 and 6q27, with none positioned within the CDR. In seven cases breakpoints clustered within a 14 Mb region of 6q22-q23 but different subregions were defined for five analysed in detail. Complex rearrangements in one case of ALL appeared to result in translocation of one homologue of 6q and deletion of the second. Detailed FISH analysis defined a translocation breakpoint falling between two PACs that contained exons of a known tumour suppressor gene, the IGF2-R. Southern blot analysis failed to confirm disruption of the IGF2-R, but an IGF2-R-MRP8 fusion transcript was cloned by RACE PCR from the patients' c-DNA. Involvement of MRP8 in the translocation remains in doubt because attempts to confirm the presence of the fusion transcript were unsuccessful.
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The promyelocytic leukaemia gene product PML interacts with Myc and influences the expression of Myc target genesCairo, Stefano January 2004 (has links)
No description available.
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DNA mismatch repair deficiency in therapy related acute myeloid leukaemia / myelodysplastic syndromeOffman, Judith Muriel January 2004 (has links)
No description available.
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Profiling of differential gene expression in acute myeloid leukaemia and the effects of stem cell factor on expression levelsCourt, Emma Louise January 2004 (has links)
No description available.
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Role and regulation of FOXO transcription factors in chronic myeloid leukaemia (CML)Hui, Chung-Yee Rosaline January 2007 (has links)
No description available.
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Targets for therapy of chronic myeloid leukaemiaCwynarski, Katherine Louise January 2004 (has links)
No description available.
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Bisphosphonates in the treatment of Imatinib-resistant chronic myeloid leukaemiaChuah, Charles Thuan Heng January 2006 (has links)
No description available.
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