MACROPHAGE ACTIVATION FOR TUMOR CELL KILLING

本计划项目竞合续签申请作为其长期 目的描述巨噬细胞参与的分子事件 激活对肿瘤细胞的杀伤。建议的研究将集中在 基因激活和基因产物共同调节 一氧化氮依赖型杀瘤药物的开发与抑制 活动，在某些情况下通过自分泌反馈。早期、中期 并将对晚期基因表达进行研究。这三个研究项目 在科学和财政上都是紧密结合在一起的。项目1，Lead 戴维·C·莫里森将研究内毒素的分子机制-- 启动的信令，导致激活和 杀瘤活性的表达。他将有三个具体目标： (I)鉴定各种内毒素受体的具体作用 分子(如p73、CD14和CD11/18)与信号转导、基因 转录和激活以杀死肿瘤细胞；(Ii)定义 内毒素介导的巨噬细胞重编程的分子基础 功能；以及(Iii)探索选择性巨噬细胞 重新编程也在体内发生，在模型肿瘤系统中呈现 对脂多糖致死作用敏感的小鼠。项目#2，导演 铃木忠雄，将研究细胞质激活的序列 由内毒素启动的事件，其中一些依赖于 巨噬细胞基因产物的自动/旁分泌反馈。两个具体目标 将继续：(I)研究内毒素触发的信号转导 导致核因子-kappa-B活化的机制，和(Ii) 研究巨噬细胞能否在体外和体外被激活 已稳定转染的肿瘤靶细胞或 用编码干扰素-β、干扰素-γ的基因进行转导， 单核细胞趋化肽(MCP-1，又名JE)或其组合 插入到真核表达载体中。项目#3，由Stephen领导 W.Russell将研究细胞核中的事件，特别是如何 脂多糖/干扰素介导的诱导型一氧化氮基因的表达 一氧化氮合酶(INOS)是受调控的。将有两个具体目标 本项目：(I)确定以下网络的组成部分 正/负向转录因子和反应元件 调节小鼠诱导型一氧化氮合酶基因表达，以及(Ii)鉴定 干扰素-β影响诱导型一氧化氮合酶产生的机制(S)，两者均为正向 也是消极的。支持服务核心组件，也将是 由罗素博士领导，旨在促进、协调和促进 这三个项目各自进行的研究。它将会遇到 三个具体需求，即(I)方案协调和 促进、(2)试剂的质量控制和标准化以及 牢房，以及(Iii)文书支助。总体而言，高度集成 建议的方法，再加上一个核心组件 积极促进研究生产力，预计将导致 组件项目之间的广泛协作，因此， 达到比当前更高的生产率 演出期间(34份全文手稿已出版或 在2年内提交)。

This Program Project competing renewal application has as its long-term objective the delineation of molecular events involved in macrophage activation for tumor cell killing. The research proposed will focus on gene activation and products of genes that collectively regulate the development and suppression of nitric oxide-dependent tumoricidal activity, in some cases through autocrine feedback. Early, intermediate and late gene expression will be studied. The three research projects are tightly integrated, both scientifically and fiscally. Project #1, led by David C. Morrison, will investigate molecular mechanisms of LPS- initiated signalling that lead to the development of activation and the expression of tumoricidal activity. He will have three specific aims: (i) to characterize the specific contributions of various LPS receptor molecules (e.g., p73, CD14, and CD11/18) to signal transduction, gene transcription, and activation for tumor cell killing; (ii) to define the molecular basis for LPS-mediated reprogramming of macrophages for altered function; and (iii) to explore the hypothesis that selective macrophage reprogramming also occurs in vivo in a model tumor system that renders mice hypersusceptible to the lethal effect of LPS. Project #2, directed by Tsuneo Suzuki, will investigate the sequence of cytoplasmic activation events that are initiated by LPS, some of which are dependent on auto/paracrine feedback of macrophage gene products. Two specific aims will be pursued: (i) to investigate LPS-triggered signal transduction mechanisms that lead to the activation of NF-kappa-B, and (ii) to investigate whether or not macrophages can be activated in vitro and in vivo by tumor target cells that have been stably transfected or transduced with the genes encoding for either IFN-beta, IFN-gamma, monocyte chemotactic peptide (MCP-1, aka JE), or combinations thereof inserted into eukaryotic expression vectors. Project #3, led by Stephen W. Russell, will investigate events in the cell nucleus, specifically how LPS/IFN-mediated expresion of the gene that encodes for inducible nitric oxide synthase (iNOS) is regulated. There will be two specific aims in this project: (i) to identify the components of the network of transcription factors and responsive elements that positively/ negatively regulate mouse iNOS gene expression, and (ii) to identify the mechanism(s) by which IFN-beta affects iNOS production, both positively and negatively. The Support Services Core Component, which will also be led by Dr. Russell, is designed to facilitate, coordinate and foster research that is conducted by each of the three projects. It will meet three specific needs, namely those for (i) program coordination and facilitation, (ii) quality control and standardization of reagents and cells, and (iii) clerical support. Overall, the highly integrated approach that is proposed, coupled with a Core Component that will proactively foster research productivity, is expected to lead to extensive collaborations between the component projects and, therefore, to even greater productivity than has characterized the current performance period (34 full length manuscripts either published or submitted in 2+ years).