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Title: Elucidating the functional role of CD38 in chronic lymphocytic leukaemia
Author: Pearce, Laurence
Awarding Body: Cardiff University
Current Institution: Cardiff University
Date of Award: 2011
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In this study, I applied a range of techniques in an attempt to enhance our knowledge of the role that CD38 plays in the pathogenesis of CLL. Investigation of a number of techniques to genetically modify the CLL cells led to the development of a lentiviral transduction system that was able to induce a marked increase in the ectopic expression of CD38 on the CLL cell surface. Subsequent molecular analysis identified changes in gene expression which may enhance disease progression. The pro-angiogenic growth factor VEGF and the DNA mismatch repair protein Msh6 were both identified as candidates for further investigation. This work also highlighted the challenges and limitations involved in using a lentiviral knock-in system and led to the design of experiments utilising CD31-expressing co-cultures to stimulate CD38 on the CLL cell surface. The CD31-expressing co-culture system induced survival within the CLL sample compared to cells incubated with the control, non-transfected co-culture. Increased proliferation was illustrated through the incorporation of BrdU and induction of the cell cycle protein Ki-67. Multi-colour flow cytometry was employed to observe the expression of surface and intracellular molecules which may be involved in CLL cell activation and signalling. Changes in the phenotype of the CLL cells were consistently observed which support the notion that these cells can be activated in vitro and can thereby enhance B-cell receptor signalling. Specifically, CD 19, CD38 and the aberrantly expressed tyrosine kinase Zap-70 were all induced following incubation with CD31-expressing co-culture. This is the first time that a lentiviral transduction system has been developed which efficiently expresses CD38 in a CLL cell population with little cell death. The work carried out in this project also highlights the importance of using co-culture to stimulate CD38 on the surface of the CLL cells in vitro. The novel findings within this project have given insight into some of the mechanisms of CD38 signalling, provided direction for future work and highlight the potential of CD38 as a therapeutic target in CLL.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available