Macrophage and Microglial Activation in Glioma-Associated Inflammation

该项目的目标是为申请人Nalin Gupta提供必要的专业知识， 独立的研究生涯在加州旧金山弗朗西斯科的基础上，他以前的 经验和培训。这将通过一个由博士监督的指导研究项目来完成。 以色列F. Charo，以及免疫学和转基因小鼠模型使用方面的额外培训。古普塔医生是一位 儿科神经外科医生，对神经肿瘤学有临床和研究兴趣。他的项目，“巨噬细胞和 神经胶质瘤相关炎症中的小胶质细胞激活1解决了脑肿瘤研究不足的方面 生物学：炎症细胞和肿瘤细胞之间的相互作用。这个项目的假设是， 巨噬细胞和小神经胶质细胞通过特异性趋化因子的过表达被募集到高级别神经胶质瘤， 单核细胞趋化蛋白-1（MCP-1），并且该过程促进肿瘤生长。具体 检验这一假设的目的是：a）确定肿瘤相关巨噬细胞的起源，B）测量 CCR 2细胞因子受体缺失对发生少突神经胶质瘤的小鼠中神经胶质瘤生长的影响， 和c）确定肿瘤和宿主来源的MCP-1对神经胶质瘤生长的贡献。的重要性 MCP-1在高级别胶质瘤中普遍存在，与浸润性巨噬细胞相关， 支持这种细胞因子在血管生成和肿瘤细胞迁移中的作用的数据。巨噬细胞的作用 和小胶质细胞对神经胶质瘤生长的影响将在转基因小鼠中使用遗传方法直接检查。小鼠 在杂合的ink 4a/arf背景中表达v-erbB癌基因可发展为高级别肿瘤， 可预测的发病率。这些肿瘤概括了人类高级别肿瘤的许多特征。为 具体目标1，动物将接受带有荧光标记细胞的骨髓移植， 可以精确地鉴定骨髓来源的细胞。细胞因子活性对神经胶质瘤生长的影响将是 通过将表达v-erbB的小鼠与缺乏CCR 2的小鼠杂交，在特定目标2中测量， MCP-1受体。将使用免疫组织化学和流式细胞术测量巨噬细胞和小胶质细胞 细胞仪最终的具体目标将使用颅内植入转化的星形胶质细胞来研究 肿瘤或宿主来源的MCP-1对肿瘤生长的影响。如果达到预期效果，我们 接下来将评估MCP-1/CCR 2信号通路的抑制剂作为潜在的抗肿瘤剂。 公共卫生相关性：伴随恶性脑肿瘤的炎症导致并发症， 患者，并可能促进肿瘤的生长。确定炎症在脑肿瘤中的作用将 为开发针对这一过程的药物提供了理论基础。这些药物的另一个可能的好处是， 还可以减少与治疗如放射相关的正常组织损伤。

The goal of this project is to provide the applicant, Nalin Gupta, with the necessary expertise to develop an independent research career at the University of California San Francisco by building on his previous experience and training. This will be accomplished through a mentored research project supervised by Dr. Israel F. Charo, and additional training in immunology and use of transgenic mouse models. Dr. Gupta is a pediatric neurosurgeon with a clinical and research interest in neuro-oncology. His project, 'Macrophage and microglial activation in glioma-associated inflammation1 addresses an understudied aspect of brain tumor biology: the interactions between inflammatory cells and tumor cells. The hypothesis of this project is that macrophages and microglia are recruited to high-grade gliomas by overexpression of a specific chemokine, monocyte chemoattractant protein-1 (MCP-1), and that this process facilitates tumor growth. The specific aims that will test this hypothesis are: a) determine the origin of tumor-associated macrophages, b) measure the effect of loss of the CCR2 cytokine receptor on glioma growth in mice developing oligodendrogliomas, and c) determine the contribution of tumor and host-derived MCP-1 on glioma growth. The importance of MCP-1 is suggested by its ubiquity in high-grade gliomas, its correlation with infiltrating macrophages, and data supporting a role for this cytokine in angiogenesis and tumor cell migration. The effect of macrophages and microglia on glioma growth will be directly examined using a genetic approach in transgenic mice. Mice expressing the v-erbB oncogene in a heterozygous ink4a/arf background develop high-grade tumors with a predictable incidence. These tumors recapitulate many of the features of human high-grade tumors. For specific aim 1, animals will receive bone marrow transplants with fluorescently labeled cells so that bone- marrow derived cells can be identified precisely. The effect of cytokine activity upon glioma growth will be measured in specific aim 2 by crossing v-erbB expressing mice with mice that lack CCR2, the cellular receptor for MCP-1. Macrophage and microglia will be measured using immunohistochemistry and flow cytometry. The final specific aim will use intracranial implantation of transformed astrocytes to study the effect of either tumor- or host-derived MCP-1 on tumor growth. If the expected results are achieved, we would next evaluate inhibitors of the MCP-1/CCR2 signaling pathway as potential anti-tumor agents. Public Health Relevance: Inflammation accompanying malignant brain tumors results in complications for patients, and may promote the growth of tumors. Identifying the role of inflammation in brain tumors would provide a rationale to develop drugs to target this process. Another possible benefit of such drugs is that normal tissue injury associated with treatments such as radiation may also be reduced.