Meningeal lymphangiogenesis effect on malignancy ofglioblastoma.

脑膜淋巴管生成对胶质母细胞瘤恶性肿瘤的影响。

• 批准号: 9758068
• 负责人: Hoyeon Eric Song
• 金额: $ 5万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-01 至 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9758068
• 关键词: Affect; Antibodies; Antigen Presentation; Antigens; Area; Autoimmunity; Autopsy; Biology; Biomedical Engineering; Brain; Brain Glioblastoma; Brain Neoplasms; CD3 Antigens; CD8B1 gene; CTLA4 gene; Cancer Immunology Science; Cells; Central Nervous System Neoplasms; Cerebrospinal Fluid; Chronic; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Code; Collaborations; Cost efficiency; Dependovirus; Development; Diagnosis; Disease; Dura Mater; Environment; Excision; Flow Cytometry; Genetic; Genome; Glioblastoma; Homeostasis; Human; Immune; Immune checkpoint inhibitor; Immune response; Immunologic Memory; Immunologic Monitoring; Immunotherapy; Implant; Individual; Inflammatory; Inflammatory Response; Kinetics; Knockout Mice; Lead; Leucocytic infiltrate; Location; Lymphangiogenesis; Lymphatic; Lymphatic System; Lymphocyte; MS4A1 gene; Malignant - descriptor; Malignant Neoplasms; Malignant neoplasm of brain; Mammals; Meningeal; Meninges; Messenger RNA; Methods; Microscopy; Modeling; Morbidity - disease rate; Mus; Neuraxis; Neurologic; Neurology; Operative Surgical Procedures; Organ; PTPRC gene; Patients; Peripheral; Population; Proliferating; Radiation therapy; Role; Sampling; Stains; Surface; System; T-Lymphocyte; Testing; Therapeutic; Time; Tumor Cell Line; United States; Up-Regulation; VEGFC gene; Vascular Endothelial Growth Factor C; Vascular Endothelial Growth Factors; Work; aggressive therapy; base; brain tissue; cell type; checkpoint inhibition; checkpoint therapy; chemotherapy; cisterna magna; cranium; design; gene induction; gene therapy; immunoreaction; implantation; improved; in vivo; insight; lymph nodes; lymphatic drainage; lymphatic vessel; mouse model; neoplastic cell; neuroimmunology; neuropathology; novel therapeutics; outcome forecast; overexpression; pre-clinical; protein expression; trafficking; transcriptome sequencing; tumor; tumor growth; tumorigenesis

Project Summary Glioblastomas are the most common form of primary malignant brain tumors and affect over 15,000 new individuals in the United States annually. With surgical resection, chemotherapy and radiotherapy the prognosis for patients is only at 15 months; and while checkpoint inhibitor therapy has provided unprecedented clinical benefit for other cancers, its effect on brain tumors is unclear. This is attributed to the unique immune environment in the central nervous system, with the draining lymphatic vasculature only being (re)identified few years ago. Before this discovery, the brain was often thought to be immune-privileged, not having traditional efferent and afferent immune cell trafficking and unable to evoke similar immune responses as peripheral organs. But even with the characterization of this CNS lymphatic network, little is known about what makes the meningeal lymphatics unique from other lymphatic networks in the body, especially during chronic inflammatory states such as the presence of a tumor. We hypothesize that at homeostasis, the CNS immunosurveillance may not be sufficient to evoke an immune response; but by increasing the lymphatic drainage to allow for more T cell priming, we can generate an adequate inflammatory response against tumors. In Aim 1, we will identify which cells are required for immunosurveillance and rejection of brain tumors. By introducing VEGFC-AAV into the cisterna magna of mice, the meningeal lymphatic network proliferates, prompting a strong immune response against orthotopic brain tumor models. With depletion antibodies and various ko mice, we will identify which cell types are required for tumor rejection in these mice through survival studies, flow cytometry and immunofluorescent staining. In Aim 2, we will investigate whether upregulation of VEGF-C can be used either as a monotherapy, or to potentiate current immunotherapy strategies. To accomplish this, we will use mRNA gene therapy to transiently increase VEGF-C in mice after brain tumor implantation. mRNA has several advantages to other gene therapy strategies-including its cost, efficiency and controlled protein expression kinetics in mammalian systems. We designed the mRNA enhancement strategy to mitigate any potential long-term harm of VEGF-C expression, while allowing VEGF-C to synergistically work with checkpoint inhibitor therapies such as, PDL-1, CTLA-4 and 4-1BB in treating GBM. Finally, in Aim 3 dural lymphatic vessels from postmortem GBM patients or from unrelated diseases will be evaluated. The dural lymphatics have only recently been identified in mice, and there are still many questions regarding its role in patients. We will evaluate what effects the chronic inflammatory state of a brain tumor has on the meningeal lymphatics by staining GBM patient’s dura (postmortem) and compared to those who died of unrelated causes. These three aims will help support our hypotheses of how immunosurveillance occurs in the CNS and help us design better treatment for GBM. We expect that our findings will uncover new strategies to make the CNS vulnerable to immunotherapy and help gain new insight to understanding cancer immunology. !!

项目摘要 胶质母细胞瘤是最常见的原发性恶性脑肿瘤， 每年在美国。通过手术切除、化疗和放疗， 患者的预后仅在15个月;虽然检查点抑制剂治疗提供了前所未有的 虽然它对其他癌症有临床益处，但它对脑肿瘤的影响尚不清楚。这是由于独特的免疫 在中枢神经系统的环境中，引流淋巴管系统仅被（重新）识别很少 年前在这一发现之前，大脑通常被认为是免疫特权的，没有传统的 传出和传入免疫细胞运输，并且不能引起与外周血类似的免疫应答。 机关但是，即使有了这种中枢神经系统淋巴网络的特征，人们对是什么使这种淋巴网络成为一种新的淋巴网络仍知之甚少。 脑膜淋巴管不同于体内其他淋巴网络，特别是在慢性 炎症状态，例如存在肿瘤。我们假设在体内平衡时，中枢神经系统 免疫监视可能不足以引起免疫反应;但通过增加淋巴细胞的数量， 引流以允许更多的T细胞引发，我们可以产生针对肿瘤的足够的炎症反应。 在目标1中，我们将确定哪些细胞是脑肿瘤免疫监视和排斥所需的。 通过将VEGFC-AAV引入小鼠的小脑延髓池，脑膜淋巴网络增殖， 促进针对原位脑肿瘤模型的强烈免疫应答。用耗尽抗体和 在各种KO小鼠中，我们将通过存活来确定这些小鼠中肿瘤排斥所需的细胞类型。 研究、流式细胞术和免疫荧光染色。在目标2中，我们将研究上调是否 VEGF-C可以用作单一疗法，或加强目前的免疫治疗策略。到 为了实现这一目标，我们将使用mRNA基因治疗来瞬时增加小鼠脑肿瘤后的VEGF-C 置入mRNA与其他基因治疗策略相比具有几个优势-包括其成本，效率和成本。 在哺乳动物系统中控制蛋白质表达动力学。我们设计了mRNA增强策略 以减轻VEGF-C表达的任何潜在的长期损害，同时允许VEGF-C协同工作， 与检查点抑制剂疗法如PDL-1、CTLA-4和4-1BB联合治疗GBM。最后，在Aim 3中 将评价来自死后GBM患者或来自无关疾病的硬脑膜淋巴管。 硬脑膜炎最近才在小鼠中被发现，关于它的作用仍有许多问题。 在患者中的作用。我们将评估脑肿瘤的慢性炎症状态对脑组织的影响。 通过对GBM患者的硬脑膜进行染色（尸检），并与死于GBM的患者进行比较， 无关的原因。这三个目标将有助于支持我们的假设，即免疫监视是如何发生的， 帮助我们设计更好的治疗GBM的方法。我们希望我们的发现将揭示新的策略， 使中枢神经系统容易受到免疫治疗的影响，并帮助获得了解癌症免疫学的新见解。 !!