Macrophage Signaling Pathways Enhancing Tumor Progression

殖民地-1（CSF-1）受体（CSF-IR）激酶指导多能造血细胞 采取巨噬细胞的命运和调节分化细胞的功能。以前的结果从这个 程序显示CSF-1驱动的肿瘤相关巨噬细胞（TAM）调节血管生成， 小鼠乳腺癌模型中癌细胞的侵袭、浸润和转移。TAM生产 血管生成因子和细胞因子，TAM迁移和TAM促进癌细胞迁移已经被 被鉴定为重要的CSF-1 R介导的事件。我们分析了巨噬细胞CSF-IR磷酸酪氨酸 （pTyr）信号通路已经显示CSF-1 R pTyr通路控制CSF-1调节的增殖， 而其它pTyr途径调节分化、形态和运动性。特别是，我们已经证明， pTyr-721通路对于巨噬细胞和癌细胞之间的旁分泌相互作用至关重要 在体外用于巨噬细胞在肿瘤内的运动，在体内用于血管生成因子的产生， 参与旁分泌回路以激活肿瘤细胞侵袭力的EGF的释放。因此我们 推测CSF-IR pTyr-721信号通路在TAM中起作用以调节TAM促进 癌细胞进展和转移。Proiect 2的总体目标是通过直接体内试验证实这一点。 分析，以确定何时以及在何处巨噬细胞增强由CSF介导的肿瘤进展- 1 R pTyr-721信号传导发生，并确定涉及的下游途径。为了验证这一 假设，我们提出了一个全面的方法，涉及三个具体的施舍。在第一个目标中，我们将 确立CSF-1 R pTyr-721信号传导在小鼠乳腺肿瘤进展中的体内作用， 转移使用新的成像方法。在第二个目标中，CSF-IR pTyr-721依赖性信号传导 分子和巨噬细胞产生的细胞因子， 将使用新开发的准确模拟旁分泌相互作用的测定法在体外进行检查， TAM介导的肿瘤细胞内渗和外渗。在第三个目标中，根据以下条件选择分子： 将检查这些体外功能测定中它们在肿瘤进展中的体内作用， 转移将通过人CDP乳腺癌的免疫组化来确定其临床意义。 癌症进展组织微阵列（TMA），并与项目5合作使用患者样本。 这些研究有望提供关于所选信号分子功能的体内数据 其作用于TAM中的CSF-IR下游信号传导途径以影响肿瘤进展和转移。 相关性（参见说明）： 拟议工作的结果将增强我们对抗炎药（例如， CSF-IR抑制剂）对癌症发病率和进展具有保护作用，并且预期将提供在癌症治疗中的应用。 关于在TAM中起作用以影响肿瘤进展的信号传导分子的功能的体内数据， 转移它们还有望导致确定人类诊断测试的新靶点， 到临床前研究测试新的治疗方法。

The colony stimulating factor-1 (CSF-1) receptor (CSF-IR) kinase instructs multipotent hematopoietic cells to adopt a macrophage fate and regulates the functions of differentiated cells. Previous results from this program have shown that CSF-1-driven tumor-associated macrophages (TAMs) regulate angiogenesis, carcinoma cell invasion, intravasation and metastasis in mouse models of breast cancer. TAM production of angiogenic factors and cytokines, TAM migration and TAM promotion of carcinoma cell migration have been identified as important CSF-1 R-mediated events. Our analysis of macrophage CSF-IR phosphotyrosyl (pTyr) signaling pathways has shown that CSF-1 R pTyr pathways control CSF-1-regulated proliferation, whereas other pTyr pathways regulate differentiation, morphology and motility. In particular, we have shown that the pTyr-721 pathway is critical for the paracrine interaction between macrophages and carcinoma cells in vitro and for macrophage motility within the tumor in vivo, for the production of angiogenic factors and for the release of EGF that participates in a paracrine loop to activate tumor cell invasiveness. Thus we hvpothesize that the CSF-IR pTyr-721 signaling pathways function in TAMs to regulate TAM promotion of carcinoma cell progression and metastasis. The overall aim of Proiect 2 is to confirm this by direct in vivo analysis, to determine when and where macrophage enhancement of tumor progression mediated by CSF- 1R pTyr-721 signaling is occurring and to identify the downstream pathways involved. To test this hypothesis, we propose a comprehensive approach involving three specific alms. In the first aim, we will establish the in vivo roles of CSF-1 R pTyr-721 signaling in mouse mammary tumor progression and metastasis using novel imaging approaches. In the second aim, CSF-IR pTyr-721-dependent signaling molecules and macrophage-produced cytokines that have been identified using combination of approaches will be examined in vitro using newly developed assays that accurately mimic the paracrine interaction and TAM-mediated tumor cell Intravasation and extravasation. In the third aim, molecules selected on the basis of these in vitro functional assays will be examined for their in vivo roles In tumor progression and metastasis. Their clinical significance will be determined by immunohistochemistry of human CDP Breast Cancer Progression Tissue Microarrays (TMA) and using patient samples in collaboration with Project 5. These studies are expected to provide in vivo data concerning the function of selected signaling molecules that act in CSF-IR downstream signaling pathways in TAMs to effect tumor progression and metastasis. RELEVANCE (See instructions): The results of the proposed work will enhance our understanding of why anti-inflammatory drugs (e.g. CSF-IR inhibitors) are protective against cancer incidence and progression and are expected to provide in vivo data concerning the function of signaling molecules that act in TAMs to effect tumor progression and metastasis. They are also expected to lead to the identification of new targets for diagnostic tests in man and to preclinical studies testing novel therapeutic approaches.