MACROPHAGE ACTIVATION FOR TUMOR CELL KILLING

本计划项目竞争性续期申请已作为其长期 目的阐明巨噬细胞介导的分子事件， 激活以杀死肿瘤细胞。 拟议的研究将侧重于 基因激活和基因产物共同调节 一氧化氮依赖性肿瘤杀伤剂的开发和抑制 活性，在某些情况下通过自分泌反馈。 早期、中期 并研究晚期基因表达。 三个研究项目 是紧密结合在一起的，无论是科学上还是精神上。项目#1，领导 由大卫C.莫里森，将研究LPS的分子机制， 启动信号传导，导致激活的发展， 表达杀肿瘤活性。 他将有三个具体目标： (i)为了表征各种LPS受体的特异性贡献， 分子（例如，p73、CD 14和CD 11/18）与信号转导、基因 转录和激活肿瘤细胞杀伤;（ii）定义 LPS介导的巨噬细胞重编程的分子基础 功能;及（iii）探讨选择性巨噬细胞 重编程也在体内发生在模型肿瘤系统中， 小鼠对LPS的致死作用过敏。 项目#2，指导 铃木恒夫，将研究细胞质激活的顺序， 由LPS引发的事件，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 将追求：（一）研究LPS触发的信号转导 导致NF-κ-B活化的机制，和（ii） 研究巨噬细胞是否可以在体外和体内被激活。 通过已经稳定转染的肿瘤靶细胞或 用编码IFN-β，IFN-γ， 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.罗素，将研究细胞核中的事件，特别是如何 LPS/IFN介导的诱导型一氧化氮合酶基因表达 氧化物合酶（iNOS）受到调节。 将有两个具体目标， 本项目：（一）确定网络的组成部分， 转录因子和应答元件， 调节小鼠iNOS基因表达，和（ii）鉴定 IFN-β影响iNOS产生的机制， 消极的。 支助事务核心部分也将 由罗素博士领导，旨在促进，协调和促进 这三个项目中的每一个都进行了研究。 它将满足 三个具体需求，即㈠方案协调和 便利化，（ii）试剂的质量控制和标准化， （三）文书支持。 总体而言，高度一体化的 提出的方法，加上一个核心组件，将 积极促进研究生产力，预计将导致 各组成项目之间的广泛合作，因此， 甚至比目前的生产力更高 业绩期间（34全文手稿出版或 2年以上）。

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).