SELF-RENEWAL IN NORMAL & LEUKEMIC HEMOPOIETIC STEM CELLS

建议研究失调的CF产生(GM-CSF， G-CSF、多粒细胞集落刺激因子)单独和联合癌基因表达 MYC、RAF/myc和bcr/abl在小鼠髓系白血病发生中的作用这个 目的是建立人髓系白血病的小鼠模型 通过在一些人身上复制疑似自分泌的脑脊液 白血病，已知的涉及癌基因的易位，尤其是 Bcr/abl与脑脊液对增殖的依赖。为了这个目的，一只鼠标 具有CM-CSF转基因的株系和含有 将使用脑脊液基因和癌基因。多潜能干细胞和 承诺的祖细胞将被MPZen(GM-CSF)感染， MPZen(多粒细胞集落刺激因子)或MPZen(粒细胞集落刺激因子)移植到致死照射的 受体或在半固体培养基中培养。体内研究将 尝试确定自分泌和/或升高的脑脊液水平 产生改变多潜能细胞(CFU-S)的增殖和/或 承诺。这些参数将通过研究个体脾进行分析。 菌落为细胞形态和CFU-S含量，感染能力 多潜能细胞在竞争性再繁殖分析中的竞争 病毒整合模式。体外研究将被进行到 确定自分泌产量是否与外源添加的 纯化的脑脊液改变承诺的祖细胞增殖和/或 差异化。在所有情况下，动物和体外培养都将是 监测以确定异常的脑脊液产生是否使细胞易感 白血病的转化。还将进行互补性研究 逆转录病毒载体感染转基因GM-CSF骨髓细胞 含有myc、raf/myc或bcr/abl。另外，正常的骨髓细胞 会感染脑脊液或癌基因逆转录病毒的各种 组合和序列。这些研究的目的是确定 自分泌脑脊液产生与癌基因表达增加相结合 髓系白血病转化。

It is proposed to study the effects of dysregulated CF production (GM-CSF, G-CSF, Multi-CSF) alone and in combination with expression of the oncogenes myc, raf/myc and bcr/abl on murine myeloid leukemia development. The objective is to develop murine models for human myeloid leukemia development by reproducing the suspected autocrine production of CSF in some human leukemias, the known translocations involving oncogenes, particularly bcr/abl and the CSF dependence for proliferation. For this purpose a mouse line having a CM-CSF transgene and recombinant retroviruses containing the CSF genes and oncogenes will be used. Multipotential stem cells and committed progenitor cells will be infected with MPZen(GM-CSF), MPZen(Multi-CSF) or MPZen(G-CSF) and transplanted into lethally-irradiated recipients or cultured in semisolid medium. The in vivo studies will attempt to determine whether autocrine and/or elevated levels of CSF production alter multipotential cell (CFU-S) proliferation and/or commitment. These parameters will be analyzed by studying individual spleen colonies for cellular morphology and CFU-S content, ability of infected multipotential cells to compete in competitive repopulation assays and by viral integration patterns. The in vitro studies will be performed to determine whether autocrine production compared to exogenous addition of purified CSF's alters committed progenitor cell proliferation and/or differentiation. In all cases, animals and in vitro cultures will be monitored to determine whether dysregulated CSF production predisposes cells to leukemic transformation. Complementation studies will also be performed by infecting transgenic GM-CSf bone marrow cells with retroviral vectors containing myc, raf/myc or bcr/abl. In addition, normal bone marrow cells will be infected with the CSF or oncogene retroviruses in various combinations and sequences. These studies aim to determine whether autocrine CSF production in combination with oncogene expression increase myeloid leukemic transformation.