SELF-RENEWAL IN NORMAL & LEUKEMIC HEMOPOIETIC STEM CELLS

正常情况下的自我更新

• 批准号: 3167120
• 负责人: DONALD METCALF
• 金额: $ 9.87万
• 依托单位: WALTER AND ELIZA HALL INST MEDICAL RES
• 依托单位国家: 美国
• 财政年份: 1983
• 资助国家: 美国
• 起止时间: 1983-06-01 至 1992-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3167120
• 关键词: bone marrow; cell differentiation; cellular oncology; clone cells; colony stimulating factor; disease /disorder model; gene complementation; genetic manipulation; genetically modified animals; hematopoietic stem cells; laboratory mouse; leukocyte activation /transformation; model design /development; molecular cloning; myelogenous leukemia; neoplasm /cancer transplantation; oncogenes; radiation immunosuppression; spleen; tissue mosaicism; viral leukemia; viral leukemogenesis

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.

建议研究 CF 产生失调（GM-CSF、 G-CSF、Multi-CSF）单独或与癌基因表达组合 myc、raf/myc 和 bcr/abl 对小鼠髓系白血病发展的影响。 这 目标是开发用于人类骨髓性白血病发展的小鼠模型 通过在某些人类身上复制疑似自分泌的脑脊液 白血病，已知涉及癌基因的易位，特别是 bcr/abl 和 CSF 增殖依赖性。 为此，鼠标 具有 CM-CSF 转基因的品系和含有 将使用脑脊液基因和癌基因。 多能干细胞和 定向祖细胞将被MPZen(GM-CSF)感染， MPZen(Multi-CSF) 或 MPZen(G-CSF) 并移植到致死辐射中 受体或在半固体培养基中培养。 体内研究将 尝试确定脑脊液是否自分泌和/或水平升高 生产改变多能细胞（CFU-S）增殖和/或 承诺。 这些参数将通过研究个体脾脏来分析 菌落细胞形态、CFU-S含量、感染能力 多能细胞在竞争性再增殖测定中进行竞争，并通过 病毒整合模式。 体外研究将进行 确定自分泌产生与外源添加相比是否 纯化的脑脊液改变定向祖细胞增殖和/或 差异化。 在所有情况下，动物和体外培养物都将 监测以确定脑脊液产生失调是否会导致细胞易感 向白血病转化。 还将进行补充研究 通过用逆转录病毒载体感染转基因 GM-CSf 骨髓细胞 包含 myc、raf/myc 或 bcr/abl。 此外，正常骨髓细胞 会在各种情况下感染脑脊液或癌基因逆转录病毒 组合和序列。 这些研究旨在确定是否 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。