SELF-RENEWAL IN NORMAL & LEUKEMIC HEMOPOIETIC STEM CELLS

正常情况下的自我更新

• 批准号: 3167125
• 负责人: DONALD METCALF
• 金额: $ 8.74万
• 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/3167125
• 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.

建议研究调节异常的CF产生（GM-CSF， G-CSF、Multi-CSF）单独和与癌基因表达联合 myc、raf/myc和bcr/abl对小鼠髓系白血病发生的影响。 的 目的是建立人类髓系白血病发展的小鼠模型 通过在某些人中复制疑似CSF的自分泌产物， 白血病，已知的涉及癌基因的易位，特别是 bcr/abl和CSF依赖性增殖。 为此， 具有CM-CSF转基因的细胞系和含有该转基因的重组逆转录病毒 将使用CSF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），并移植到经致死照射的 受体或在半固体培养基中培养。 体内研究将 试图确定是否自分泌和/或CSF水平升高 产生改变多能细胞（CFU-S）增殖和/或 承诺. 这些参数将通过研究个体脾脏进行分析 集落的细胞形态和CFU-S含量，感染能力 多能细胞在竞争性再增殖试验中竞争， 病毒整合模式 将进行体外研究， 确定是否自分泌生产相比，外源性添加 纯化的CSF改变定向祖细胞增殖和/或 分化 在所有情况下，动物和体外培养物将被 监测以确定CSF产生失调是否使细胞 转化为白血病 还将进行互补研究 通过用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。此外，正常骨髓细胞 将感染CSF或癌基因逆转录病毒， 组合和序列。 这些研究旨在确定 自分泌CSF产生与癌基因表达增加相结合 骨髓性白血病转化