THE ROLE OF CSF-1 MEDIATED MACROPHAGE CHEMOTAXIS IN CARCINOMA CELL INVASION

CSF-1介导的巨噬细胞趋化作用在癌细胞侵袭中的作用

• 批准号: 7345406
• 负责人: Dianne Cox
• 金额: $ 30.63万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-02-01 至 2011-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7345406
• 关键词: 1-Phosphatidylinositol 3-Kinase; Actins; Area; Biochemical; Biological Assay; Biosensor; Cancer Biology; Carcinoma; Cells; Chemotactic Factors; Chemotaxis; Complex; Cytoskeleton; Data; Detection; EGF gene; Extravasation; Feedback; Fluorescence Resonance Energy Transfer; Guanine Nucleotide Exchange Factors; Imaging Techniques; Intervention; Invasive; Investigation; Lead; Life; Macrophage Colony-Stimulating Factor; Maintenance; Malignant Epithelial Cell; Mediating; Movement; Mutation; Neoplasm Metastasis; Numbers; Phenotype; Phosphatidylinositide 3-Kinase Inhibitor; Phosphoinositide-3-Kinase, Catalytic, Gamma Polypeptide; Play; Positive Reinforcements; Process; Proteins; Resolution; Role; Site; Small Interfering RNA; Solid Neoplasm; Source; Technology; Tumor Cell Invasion; Wiskott-Aldrich Syndrome; base; cell motility; cellular imaging; extracellular; in vitro Assay; macrophage; neoplastic cell; novel; paracrine; polymerization; prevent; protein activation; reconstitution; response; rho GTP-Binding Proteins; therapeutic target; tumor

Tumor-associated macrophages (TAM), which are present in large numbers in many tumors, appear to play an important role in promoting the progression of solid tumors to an invasive, metastatic phenotype. It has been shown recently that TAM participate in a paracrine loop with carcinoma cells such that the CSF-1- producing carcinoma cells and the EGF-secreting macrophages (MD) interact to promote mutual chemotaxis leading to invasion and extravasation of carcinoma cells. In MDs, PI 3-kinase, Cdc42 and WASP (Wiskott- Aldrich syndrome protein) are required for migrating cells to detect the source of a chemoattractant. It has been speculated that amplification of the extracellular gradient occurs through an intracellular positive feedback loop involving PI 3-kinase, Rho GTPases and actin assembly. The precise function of WASP in gradient detection is not known. Based on preliminary data, WASP activity is required for CSF-1 induced actin polymerization in MDs which may contribute to the reinforcement of the positive feedback loop in CSF- 1 gradient detection (chemotactic sensing) leading to efficient chemotaxis. In the first Specific Aim, the role of PI3K and Cdc42 in the activation of WASP will be determined using PI3K inhibitors, CSF-1R mutations that lack PI3K activation, and by reducing endogenous levels of Cdc42 using siRNA technology. The mechanisms of WASP activation will be examined through mutational analysis of a fluorescence resonance energy transfer (FRET) based WASP biosensor. In Specific Aim 2, the role of WASP mediated actin polymerization in chemotactic sensing will be determined. The role of WASP in the localization of PI3K and Cdc42 in chemotactic sensing will be determined by live cell imaging. In addition, we will perform biochemical isolation of CSF-1 elicited pseudopods from wild-type and WASP-deficient macrophages in order to identify potential WASP interacting proteins. In Specific Aim 3 The effect of WASP on polarization and movement of MD and carcinoma cells will be examined using an in vitro assay that reconstitutes the paracrine interaction between MD and tumor cells. Relevance: Understanding how MDs migrate into a tumor site and their interaction with carcinoma cells, is an important area of investigation in cancer biology. Since WASP is specifically required for MD chemotaxis it may represent a novel target and may lead to new therapies to prevent metastasis

肿瘤相关巨噬细胞 (TAM) 在许多肿瘤中大量存在，似乎发挥着重要作用 在促进实体瘤进展为侵袭性、转移性表型方面发挥着重要作用。它有 最近的研究表明，TAM 参与癌细胞的旁分泌环路，使得 CSF-1- 产生癌细胞和分泌 EGF 的巨噬细胞 (MD) 相互作用以促进相互趋化 导致癌细胞的侵袭和外渗。在 MD 中，PI 3-激酶、Cdc42 和 WASP (Wiskott- 奥尔德里奇综合征蛋白）是迁移细胞检测化学引诱剂来源所必需的。它有 据推测，细胞外梯度的放大是通过细胞内的正向梯度发生的。 反馈环路涉及 PI 3 激酶、Rho GTPases 和肌动蛋白组装。 WASP 的精确功能 梯度检测是未知的。根据初步数据，CSF-1 诱导需要 WASP 活性 MD 中的肌动蛋白聚合可能有助于强化 CSF 中的正反馈回路 1 梯度检测（趋化传感）可实现有效的趋化作用。在第一个具体目标中，角色 PI3K 和 Cdc42 在 WASP 激活中的作用将使用 PI3K 抑制剂、CSF-1R 突变来确定 缺乏 PI3K 激活，并使用 siRNA 技术降低 Cdc42 的内源水平。这 WASP 激活机制将通过荧光共振突变分析进行检查 基于能量转移（FRET）的 WASP 生物传感器。在具体目标 2 中，WASP 介导的肌动蛋白的作用 将确定趋化传感中的聚合。 WASP 在 PI3K 和本地化中的作用 趋化传感中的 Cdc42 将通过活细胞成像来确定。此外，我们还将表演 CSF-1 的生化分离从野生型和 WASP 缺陷型巨噬细胞中诱导出伪足 为了识别潜在的 WASP 相互作用蛋白。具体目标 3 WASP 对极化的影响 MD 和癌细胞的运动将使用体外测定进行检查，该测定可重建 MD和肿瘤细胞之间的旁分泌相互作用。 相关性：了解 MD 如何迁移到肿瘤部位以及它们与癌细胞的相互作用是重要的 癌症生物学研究的一个重要领域。由于 MD 趋化性特别需要 WASP 它可能代表一个新的靶点，并可能导致预防转移的新疗法