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

基本信息

批准号：
7569444
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
项目类别：
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/7569444
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 Investigation Lead Life Macrophage Colony-Stimulating Factor Maintenance Malignant Epithelial Cell Mediating Movement Mutation Neoplasm Metastasis Phenotype Phosphatidylinositide 3-Kinase Inhibitor 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

项目摘要

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

在许多肿瘤中大量存在的肿瘤相关巨噬细胞（TAM）似乎在玩耍在促进实体瘤向侵入性转移表型的进展方面的重要作用。它有最近显示，TAM参与了带有癌细胞的旁分泌环，因此CSF-1-- 产生癌细胞和EGF分泌巨噬细胞（MD）相互作用以促进相互趋化性导致癌细胞的侵袭和渗出。在MDS中，PI 3-激酶，Cdc42和黄蜂（Wiskott- Aldrich综合征蛋白是迁移细胞需要检测趋化剂来源所必需的。它有被推测，细胞外梯度的扩增是通过细胞内阳性发生的反馈回路涉及PI 3-激酶，Rho GTPases和肌动蛋白组件。黄蜂的精确功能梯度检测尚不清楚。基于初步数据，CSF-1诱导的WASP活动需要 MDS中的肌动蛋白聚合可能有助于加强CSF- 1梯度检测（趋化感应）导致有效的趋化性。在第一个特定目标中，角色在黄蜂激活中PI3K和CDC42的激活中，将使用PI3K抑制剂确定CSF-1R突变缺乏PI3K激活，并且通过使用siRNA技术降低内源性CDC42。这黄蜂激活的机制将通过突变分析荧光共振基于能量转移（FRET）WASP生物传感器。在特定的目标2中，黄蜂介导的肌动蛋白的作用将确定趋化感应中的聚合。黄蜂在PI3K定位中的作用趋化感应中的Cdc42将由活细胞成像确定。此外，我们将表演 CSF-1的生化分离从野生型和黄蜂缺乏巨噬细胞引起的假脚架中为了识别潜在的黄蜂相互作用蛋白。在特定的目标3中，黄蜂对极化的影响 MD和癌细胞的运动将使用重构的体外测定法检查 MD和肿瘤细胞之间的旁分泌相互作用。相关性：了解MDS如何迁移到肿瘤部位及其与癌细胞的相互作用是癌症生物学研究的重要领域。由于MD趋化性是特别需要黄蜂的它可能代表一个新颖的目标，并可能导致新的疗法以防止转移