The oncogenic functions of Neuropilin-1 in the glioblastoma tumor microenvironment

Neuropilin-1 在胶质母细胞瘤肿瘤微环境中的致癌功能

• 批准号: 10441240
• 负责人: Daniel Radin
• 金额: $ 3.71万
• 依托单位: STATE UNIVERSITY NEW YORK STONY BROOK
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10441240
• 关键词: Ablation; Adopted; Adult; Affect; Angiogenic Factor; Anti-Inflammatory Agents; Area; Behavior; Binding; Breast Cancer Model; Cell Proliferation; Cells; Central Nervous System Neoplasms; Coculture Techniques; Complex; Conditioned Culture Media; Data; Fostering; Genes; Genetic Transcription; Glioblastoma; Glioma; Goals; Growth; Hypoxia; Immune; Immune Evasion; Immunocompetent; Immunosuppression; Innate Immune System; Knock-out; Knowledge; Laboratories; Malignant neoplasm of lung; Malignant neoplasm of pancreas; Mediating; Mediator of activation protein; Methods; Microglia; Mus; Myelogenous; Myeloid Cells; Nature; Neuropilin-1; Oncogenic; Operative Surgical Procedures; Output; Oxygen; Pathway interactions; Phenotype; Primary Neoplasm; Process; Property; Quantitative Reverse Transcriptase PCR; Radiation; Reporting; Research; Resistance; Role; Shapes; Signal Pathway; Signal Transduction; Testing; Transforming Growth Factor beta; Tumor Debulking; Vascular Endothelial Growth Factors; Work; angiogenesis; anti-cancer; chemokine; chemoradiation; chemotherapy; cytokine; erbB-2 Receptor; exhaust; experimental study; gene function; glioma cell line; in vivo; inflammatory marker; inhibitor; macrophage; mouse model; neoplastic cell; nerve stem cell; normoxia; novel therapeutics; phosphoproteomics; receptor; recruit; release factor; response; standard of care; stem cell biomarkers; stem cell growth; stem cell proliferation; stem cells; stemness; trafficking; tumor; tumor growth; tumor hypoxia; tumor initiation; tumor microenvironment; tumor-immune system interactions; tumorigenic

Abstract Glioblastoma (GB) is the most common primary tumor of the central nervous system (CNS) in adults. Despite aggressive surgery, radiation, and chemotherapy, median survival is only 15-20 months. In GB tumors, approximately 30-50% of the cells in the tumor are innate myeloid immune cells, primarily macrophages (MP) and microglia (MG). Research in the Tsirka laboratory has shown that MP and MG enhance GB growth by promoting angiogenesis and fostering an immunosuppressive tumor microenvironment (TME), partly through the action of the co-receptor Neuropilin-1 (NRP1). Our recent work has suggested distinct roles for MG and MP in glioma. Expression of NRP1 on glioma-associated MP and MG (GAM) is critical for their oncogenic activity. Deletion of NRP1 from myeloid cells blunts angiogenesis and results in decreased GB volume. Prior research has shown that hypoxic glioma cells upregulate stem cell-associated genes and function as glioma stem cells (GSC), which are thought to drive the immunosuppressive and angiogenic nature of GAM. NRP1 has been shown to be critical for the GAM responses to hypoxia. The long-term goal of our research group is to define the TME roles of MG and MP as well as methods through which MP and MG can be repolarized to exert anti- tumorigenic effects rather than facilitate tumor growth. The overall objective of my proposal is to determine the mechanisms by which MG and MP interact with hypoxic regions and GSC to exert oncogenic effects. I hypothesize that NRP1 activation on myeloid cells is an important mediator for pro-tumorigenic signaling between hypoxic glioma cells, GSC and GAM. To test this hypothesis, I will pursue two specific aims: 1) Determine the NRP1 pathways activated in GAM by hypoxic glioma cells; and 2) identify how GAM-NRP1 signaling affects the interaction between GSC and GAM. For the first aim, MP and MG will be separately treated with media from glioma cells cultured in normoxia or hypoxia, with or without a NRP1 inhibitor, and assessed for differences in the expression of pro/anti-inflammatory and angiogenesis factors identified through phosphoproteomics. Using immunocompetent mouse models of GB, I will investigate how GAM-NRP1 affects GAM phenotypes and tumor cell proliferation in normoxic and hypoxic tumor regions. In the second aim, the contribution of GAM-NRP1 to GSC/GAM interactions will be interrogated in culture and in vivo. The proposed research is expected to be significant because it will determine roles for MP and MG behavior in the context of hypoxic areas of glioma and further explore the interaction between GSC and GAM. It will also help determine whether modulation of NRP1 is a viable method for promoting anti-tumorigenic outputs for GAM.

摘要 胶质母细胞瘤(GB)是成人中枢神经系统最常见的原发肿瘤。尽管 积极的手术、放疗和化疗，中位生存期只有15-20个月。在GB肿瘤中， 大约30-50%的肿瘤细胞是先天的髓系免疫细胞，主要是巨噬细胞。 和小胶质细胞(MG)。齐尔卡实验室的研究表明，MP和MG通过以下方式促进GB的生长 促进血管生成和培育免疫抑制的肿瘤微环境(TME)，部分通过 共受体Neuropilin-1(Nrp1)的作用我们最近的研究表明，MG和MP扮演着不同的角色 在神经胶质瘤中。Nrp1在胶质瘤相关的MP和MG(GAM)上的表达是其致癌活性的关键。 从髓系细胞中删除Nrp1会钝化血管生成，导致GB体积减少。前期研究 研究表明，低氧脑胶质瘤细胞上调干细胞相关基因，并发挥脑胶质瘤干细胞的功能 (GSC)，它们被认为驱动GAM的免疫抑制和血管生成特性。Nrp1已经被 被证明是GAM对低氧反应的关键。我们研究小组的长期目标是定义 MG和MP的作用以及MP和MG可通过复极化发挥反作用的方法 而不是促进肿瘤的生长。我的建议的总体目标是确定 MG和MP与缺氧区和GSC相互作用发挥致癌作用的机制。我 假设髓系细胞上Nrp1的激活是促肿瘤信号的重要中介 在低氧脑胶质瘤细胞、GSC和GAM之间。为了验证这一假设，我将追求两个具体目标：1) 确定缺氧胶质瘤细胞激活的GAM中的Nrp1通路；以及2)确定GAM-Nrp1是如何 信号转导影响GSC和GAM之间的相互作用。对于第一个目标，MP和MG将分开处理 用在常氧或低氧条件下培养的胶质瘤细胞的培养液，加或不加Nrp1抑制剂，评估 促/抗炎和血管生成因子的表达差异通过 磷酸蛋白质组学。利用免疫活性的GB小鼠模型，我将研究GAM-Nrp1是如何影响 常氧区和缺氧区GAM表型与肿瘤细胞增殖在第二个目标中， GAM-Nrp1在GSC/GAM相互作用中的作用将在培养和体内进行询问。建议数 预计研究将具有重要意义，因为它将确定MP和MG行为在 并进一步探讨GSC和GAM之间的相互作用。它还将有助于确定 调节Nrp1是否是促进GAM抗肿瘤产出的可行方法。