Roles of the Immune Cytokine Environment on Clonal Growth of Murine and Human Tet2 Mutant Bone Marrow Progenitors: Implications for Myelodysplastic Syndromes

TET2 is a tumor suppressor gene that has been implicated in the epigenetic regulation of gene expression. Inactivating TET2 mutations are common in MDS. These mutations may contribute to early clonal dominance and myeloid transformation, although the exact mechanisms remain to be elucidated. Common to the environment of MDS are elevations in cytokines, such as TNFα and IFN-γ. It was hypothesized that inflammatory cytokines TNF-α and IFN-γ may promote clonal expansion of TET2 mutant progenitors. Adult (10-14 weeks-old) Tet2 wild type (+/+) and Tet2 mutant (-/-) C57BL/6 mice strains were chosen as a model system. Lineage negative cells (Lin-), enriched for hematopoietic stem and progenitor cells, were isolated from Tet2 +/+ and -/- bone marrow and cultured in the absence or presence of varying concentrations of TNFα or IFN-γ in methylcellulose colony formation assays and long term cell culture assays, over a period of 12 and 30 days respectively, and their colony growth, cell count, immunophenotype and resistance to apoptosis were examined. Where indicated, serial re-plating was performed. Expression of apoptotic regulators was assessed by qRT-PCR. In the triplicate experiments, starting with equal densities of Tet2 +/+ and -/- Lin- cells, Tet2 -/- Lin- cells displayed increased resistance to cytokine-induced growth suppression and superior colony forming ability over +/+ in the serial re-plating assays under stress of increasing TNFα or IFN γ. Tet2 -/- progenitors also displayed a lower apoptotic index compared to +/+ under stress of increasing TNFα, suggesting increased resistance to TNFα induced apoptosis. Transcriptional data showed low expression of Tnfr1, Fas and caspase 8, as well as a high expression of Bcl-2 and Iap1 in Tet2 -/- compared to +/+ under stress of TNFα. Tet2-/- also showed increased basal expression of endogenous TNFα mRNA compared to +/+. In the human colony growth assay, the clonal growth of TET2 mutant CFU-GM progenitors was enhanced at low TNFα concentrations. Conclusion: Mutations that promote resistance to environmental stem cell stressors are a known mechanism of clonal selection in aplastic anaemia and JAK2-mutant MPN and our findings suggest that this mechanism may be critical to clonal selection and dominance in MDS.

A NEW DC UPS FOR DC POWER DISTRIBUTION SYSTEM IN DATA CENTER

In recent years, the 380V DC and 48V DC distribution systems have been extensively studied for the latest data centers. It is widely believed that the 380V DC system is a very promising candidate because of its lower cable cost compared to the 48V DC system. However, previous studies have not adequately addressed the low reliability issue with the 380V DC systems due to large amount of series connected batteries. In this thesis, a quantitative comparison for the two systems has been presented in terms of efficiency, reliability and cost. A new multi-port DC UPS with both high voltage output and low voltage output is proposed. When utility ac is available, it delivers power to the load through its high voltage output and charges the battery through its low voltage output. When utility ac is off, it boosts the low battery voltage and delivers power to the load form the battery. Thus, the advantages of both systems are combined and the disadvantages of them are avoided. High efficiency is also achieved as only one converter is working in either situation. Details about the design and analysis of the new UPS are presented. For the main AC-DC part of the new UPS, a novel bridgeless three-level single-stage AC-DC converter is proposed. It eliminates the auxiliary circuit for balancing the capacitor voltages and the two bridge rectifier diodes in previous topology. Zero voltage switching, high power factor, and low component stresses are achieved with this topology. Compared to previous topologies, the proposed converter has a lower cost, higher reliability, and higher efficiency. The steady state operation of the converter is analyzed and a decoupled model is proposed for the converter. For the battery side converter as a part of the new UPS, a ZVS bidirectional DC-DC converter based on self-sustained oscillation control is proposed. Frequency control is used to ensure the ZVS operation of all four switches and phase shift control is employed to regulate the converter output power. Detailed analysis of the steady state operation and design of the converter are presented. Theoretical, simulation, and experimental results are presented to verify the effectiveness of the proposed concepts.