CYTOKINE SIGNALING IN MYELOID LEUKEMIAS

描述：（改编自申请人的摘要）本申请寻求 以表征增殖信号通路中的关键步骤， 在原始骨髓细胞中由GM-CSF激活相关细胞因子。在 至少有一些髓性白血病病例可能是由于过度活动引起的 这样的信号通路。 确定关键或独特的步骤， 这种级联反应可能允许开发药物干预， 专门破坏这些通路。 这个问题的具体假设 应用是PIM-1基因工程，一种33 kD 丝氨酸/苏氨酸激酶，是有效增殖所必需的， 通过GM-CSF和相关受体进行信号传导。 申请者还 假设pim-1的表达是一个特异性的指标， 激活这些途径。 该假设将由以下人员进行调查： 构建允许过表达或阻断的细胞系统 pim-1活性。 因子依赖性骨髓细胞系将是 用质粒构建体或逆转录病毒构建体转染， pim-1基因产物的自主表达。然后， 检查是否存在非因子依赖性生长，分化受损， 抑制细胞凋亡和同型聚集。 相似细胞系 将利用含有反义PIM-1构建体的质粒产生 以抑制内源性PIM-1基因的活性。 这些细胞将 研究了对GM-CSF和相关细胞因子及其 分化倾向。

DESCRIPTION: (Adapted from applicant's abstract)This application seeks to characterize critical steps in the proliferative signalling pathways activated by GM-CSF in related cytokines in primitive myeloid cells. At least some cases of myeloid leukemia likely result from over activity of such signal pathways. Identification of critical or unique steps in such cascades may allow development of pharmacologic interventions to specifically disrupt these pathways. The specific hypothesis of this application is that expression of the pim-1 gene project, a 33 kd serine/threonine kinase, is necessary for effective proliferative signalling through GM-CSF and related receptors. The applicant further hypothesizes that pim-1 expression serves as a specific indicator of activation of such pathways. The hypothesis will be investigated by constructing cell systems which allow for the over expression or blockade of pim-1 activity. Factor-dependent myeloid cell lines will be transfected with plasmid constructs or retroviral constructs to allow autonomous expression of the pim-1 gene product.These lines will then be examined for factor- independent growth, impaired differentiation, inhibition of apoptosis and homotypic aggregation. Similar cell lines will be created utilizing plasmids containing antisense pim-1 constructs to inhibit activity of the endogenous pim-1 gene. These cells will be studied for the responsiveness to GM-CSF and related cytokines and their propensity to differentiation.