Exosomal TRAF2-CSN5 complex mediated inflammation promotes tumor growth

外泌体TRAF2-CSN5复合物介导的炎症促进肿瘤生长

• 批准号: 8212755
• 负责人: HUANG-GE ZHANG
• 金额: --
• 依托单位: LOUISVILLE VA MEDICAL MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2015-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8212755
• 关键词: A/J Mouse; Acceleration; Anti-Inflammatory Agents; Bone Marrow; Bone Marrow Cells; Cancer Patient; Carcinogens; Cell model; Cells; Chronic; Complex; Curcumin; Data; Development; Epithelial; Epithelial Cells; Goals; Growth; Human; IL8 gene; ITGAM gene; Inflammation; Inflammatory Response; Interleukin-6; Knock-out; Lead; Link; Lung; Lung Neoplasms; Macrophage Activation; Malignant neoplasm of lung; Mediating; Membrane; Modeling; Molecular; Mus; Myelogenous; Myeloid Cells; NOD/SCID mouse; Physiologic pulse; Play; Prevention; Preventive; Production; Property; Proteins; Research; Role; SCID Mice; Small Interfering RNA; Structure of parenchyma of lung; TRAF2 gene; Testing; Therapeutic; Tissues; Tumor Necrosis Factor-alpha; Tumor Promotion; Tumor Tissue; Tumor-Derived; Ubiquitination; Urethane; Vascular Endothelial Growth Factors; Vesicle; base; cancer cell; cell growth; clinically relevant; healthy volunteer; human TNF protein; in vivo; innovation; lung Carcinoma; macrophage; migration; mouse model; neoplastic cell; novel; peripheral blood; precursor cell; public health relevance; tumor; tumor growth; ubiquilin

DESCRIPTION (provided by applicant): The overall goal of this proposal is to elucidate the mechanisms by which lung tumor exosomes promote an inflammatory response, namely the activation of macrophages, and promote the progression and growth of lung tumors. Our preliminary data indicate that pretreatment of A/J mice with exosomes produced by TC-1 lung tumor cells or exosomes isolated from the lung tissue of A/J mice that have been pretreated with the carcinogen urethane results in more rapid tumor growth and earlier progression of lung carcinomas. Urethane treatment results in the recruitment of the activated form of TRAF2 to the exosomes, and the exosomes with activated TRAF2 are taken up by the bone marrow-derived precursors of macrophages, leading to their maturation and subsequent migration to the lung. The recruitment of the activated TRAF2 to the exosomes requires degradation of the inflammation suppressor protein, CYLD, in the tumor cells; moreover, TRAF2 can mediate ubiquination of CYLD thereby promoting its degradation. The results of siRNA TRAF2 knockout indicated, however, that TRAF2 is required but is not sufficient for ubiquitination of CYLD. We have now identified two exosomal proteins that interact with TRAF2, Itch and CSN5, which have the potential to enhance the ubiquitination of CYLD in lung tumor cells. Our recent data indicating that CYLD is not packed in exosomes isolated from the peripheral blood of lung cancer patients although it is present in exosomes from healthy volunteers indicate the potential clinical relevance of this model that directly links the effects of a carcinogen with an inflammatory response and promotion of tumor development. We propose to: (1) Confirm the role of lung tumor exosomal TRAF2 in the promotion of urethane-induced lung carcinomas by determining whether knockout of TRAF2 in TC-1 lung tumor cells is sufficient for (i) inhibition of macrophage differentiation and (ii) prevention of lung tumor exosome-mediated enhancement of urethane induced lung cancer. (2) Determine if other exosomal proteins that interact with CYLD participate in the ubiquitination of CYLD in TC-1 tumor cells and determine if the elimination of these proteins stabilizes exosomal CYLD and suppresses exosomal TRAF2 activation resulting in the inhibition of macrophage activation and prevention of tumor growth. (3) Determine if macrophages pulsed with lung tumor exosomes isolated from lung cancer patients promote lung tumor growth in a NOD-SCID mouse model. The data generated should permit the development of a highly innovative model of the cellular and molecular basis for exosome-mediated chronic inflammation and promotion of tumor growth and indicate novel preventive and therapeutic strategies. PUBLIC HEALTH RELEVANCE: The proposed research is highly relevant to characterizing the association of lung tumor exosomes mediated inflammatory responses and lung tumor growth in that it is focused on characterizing the molecular mechanisms underlying the ability of tumor exosomes to both activate macrophages and promote tumor growth. The central hypothesis is that exosomes produced by lung tumor cells, or lung tumor tissue are taken up by bone marrow myeloid precursor cells. These cells further differentiate into macrophages and migrate into the lung, promote the progression of urethane- induced lung cancer.

描述（由申请人提供）： 该提案的总体目标是阐明肺肿瘤外泌体促进炎症反应（即巨噬细胞活化）并促进肺肿瘤进展和生长的机制。我们的初步数据表明，用TC-1肺肿瘤细胞产生的外泌体或从用致癌物氨基甲酸乙酯预处理的A/J小鼠肺组织中分离出的外泌体预处理A/J小鼠，会导致肿瘤生长更快，肺癌进展更早。氨基甲酸酯处理导致激活形式的 TRAF2 被招募到外泌体中，并且具有激活 TRAF2 的外泌体被骨髓来源的巨噬细胞前体吸收，导致其成熟并随后迁移到肺部。将激活的 TRAF2 招募到外泌体需要降解肿瘤细胞中的炎症抑制蛋白 CYLD；此外，TRAF2可以介导CYLD的泛素化，从而促进其降解。然而，siRNA TRAF2 敲除的结果表明，TRAF2 对于 CYLD 的泛素化来说是必需的，但还不够。我们现在已经鉴定出两种与 TRAF2 相互作用的外泌体蛋白：Itch 和 CSN5，它们有可能增强肺肿瘤细胞中 CYLD 的泛素化。我们最近的数据表明，尽管 CYLD 存在于健康志愿者的外泌体中，但从肺癌患者外周血中分离出的外泌体中并未包含 CYLD，这表明该模型的潜在临床相关性，将致癌物的影响与炎症反应和促进肿瘤发展直接联系起来。我们建议：（1）通过确定TC-1肺肿瘤细胞中TRAF2的敲除是否足以（i）抑制巨噬细胞分化和（ii）预防肺肿瘤外泌体介导的乌拉坦诱导的肺癌增强，确认肺肿瘤外泌体TRAF2在促进乌拉坦诱导的肺癌中的作用。 (2)确定与CYLD相互作用的其他外泌体蛋白是否参与TC-1肿瘤细胞中CYLD的泛素化，并确定消除这些蛋白是否可以稳定外泌体CYLD并抑制外泌体TRAF2活化，从而抑制巨噬细胞活化并预防肿瘤生长。 (3) 确定用从肺癌患者中分离出的肺肿瘤外泌体脉冲的巨噬细胞是否会促进 NOD-SCID 小鼠模型中的肺肿瘤生长。产生的数据应该允许开发外泌体介导的慢性炎症和促进肿瘤生长的细胞和分子基础的高度创新模型，并表明新的预防和治疗策略。 公共卫生相关性： 拟议的研究与表征肺肿瘤外泌体介导的炎症反应和肺肿瘤生长之间的关联高度相关，因为它的重点是表征肿瘤外泌体激活巨噬细胞和促进肿瘤生长的能力的分子机制。中心假设是肺肿瘤细胞或肺肿瘤组织产生的外泌体被骨髓髓样前体细胞吸收。这些细胞进一步分化为巨噬细胞并迁移到肺部，促进乌拉坦诱导的肺癌的进展。