THERAPEUTIC USES FOR HUMAN GRANULOCYTE CSF

人粒细胞 CSF 的治疗用途

• 批准号: 3188843
• 负责人: MICHAEL B LILLY
• 金额: $ 12.65万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-08-01 至 1992-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3188843
• 关键词: acute leukemia; cell bank /registry; colony stimulating factor; cyclophosphamide; cytosine arabinoside; doxorubicin; drug metabolism; gene expression; granulocyte; human subject; human tissue; laboratory mouse; methotrexate; myelogenous leukemia; neoplasm /cancer genetics; neoplastic cell; nucleic acid hybridization; oncogenes; osteoclast activating factor; radiotracer; transfection; xenotransplantation

The proposed studies seek to establish an experimental basis for the use of human granulocyte colony-stimulating factor (hG-CSF) in the management of patients with malignancies, particularly acute myelogenous leukemia. This factor is unique in that with in vitro systems, it can stimulate production of normal human and murine myeloid cells, and also inhibit the growth (possibly by inducing differentiation) of some human and murine acute myeloid leukemia (AML) cells. The recent cloning of a gene for hG-CSF, and its expression in vitro has led to the availability of recombinant factor, which is required for these studies. Three major specific aims are identified: 1) A new G-CSF gene from the LD1 melanoma line will be characterized by sequencing, and it will be expressed in both mammalian and bacterial expression systems to allows characterization of its protein product, and comparison with other G-CSFs. 2) recombinant hG- CSF will be used to study its supperssive or stimulatory effects on growth of human AML cells lines (PLB-985, KG-1) and fresh AML blasts from patients, growing as xenografts or in diffusion chambers in nude mice. Supporting studies will define the pharmacokinetics of rhG-CSF in mice, examine possible toxicity including osteopenia due to oseoclast activiation, and develop in vitro screening assays to predict the effect of rhG-CSF, on AML cells in vivo. These latter tests will include in vitro culture with G-CSF, binding of 125I-labelled G-CSF to AML cells, oncogene expression, and autocrine production of CSFs. Synergy between rhG-CSF and other differentiating agents (RA, 1,25-OH-VD3, mithramycin, adriamycin) will also be sought both in vitro and in vivo. A G-CSF-resistance AML subline (PLB-2R and KG-la) will be transfected with the G-CSF gene via a retroviral vector to characterize the mechanisms of G-CSF resistance in AML blasts. 3) The ability of G-CSF to modify the in vivo myelotoxicity, and to improve the therapeutic index of cytotoxic drugs used in leukemia treatment will also be examined. These studies will be used both human (PLB985, KG-1) AML lines in nude mice, and murine tumor lines (L1210, Dunn osteosarcoma). A specrum of phase specific and non-specific cytotoxic drugs (MTX, ADR, CTX, AraC) will be administrated with, or after rhG-CSF to mice.

拟议的研究旨在为 人类粒细胞菌落刺激因子（HG-CSF）的使用 在患有恶性肿瘤的患者中，特别是 急性脊髓性白血病。 这个因素是独一无二的 体外系统，它可以刺激正常人和 鼠髓样细胞，也抑制生长（可能是由 诱导某些人和鼠急性髓样的差异化） 白血病（AML）细胞。 HG-CSF基因的最近克隆， 它在体外的表达导致了 重组因子，这些研究是必需的。 确定了三个主要的特定目标：1）新的G-CSF基因 从LD1黑色素瘤系中，将以测序为特征， 它将在哺乳动物和细菌中表达 表达系统允许表征其蛋白质 产品，并与其他G-CSF进行比较。 2）重组Hg- CSF将用于研究其晚餐或刺激作用 人AML细胞系（PLB-985，kg-1）和新鲜AML的生长 患者的爆炸，作为异种移植或扩散生长 裸鼠中的钱伯斯。 支持研究将定义 小鼠RHG-CSF的药代动力学，检查可能的毒性 包括因osocolast opitigation引起的骨质减少症，并在 体外筛选测定以预测RHG-CSF对AML的影响 细胞体内。 这些后一种测试将包括与 G-CSF，125i标记的G-CSF与AML细胞的结合，癌基因 CSF的表达和自分泌产生。 之间的协同作用 RHG-CSF和其他区分剂（RA，1,25-OH-VD3， Mithramycin，Adrimycin）也将在体外和 体内。 G-CSF抵抗AML Subline（PLB-2R和KG-LA）将 通过逆转录病毒载体转染G-CSF基因 表征AML爆炸中G-CSF抗性的机制。 3）G-CSF修改体内骨髓毒性的能力和 改善用于细胞毒性药物的治疗指数 还将检查白血病治疗。 这些研究将是 在裸鼠中使用了人类（PLB985，kg-1）AML线， 鼠肿瘤系（L1210，Dunn骨肉瘤）。 一个 相特异性和非特异性细胞毒性药物（MTX，ADR，CTX，， ARAC）将通过RHG-CSF进行管理，或在鼠标上。