Brain Endothelial TNF-R1 Can Function to Inhibit Angiogenesis

脑内皮 TNF-R1 可以抑制血管生成

• 批准号: 8100461
• 负责人: Candece L Gladson
• 金额: $ 35.55万
• 依托单位: CLEVELAND CLINIC LERNER COM-CWRU
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-01 至 2014-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8100461
• 关键词: 3-Dimensional; Apoptosis; Apoptotic; Area; Binding; Biological Markers; Biopsy; Biopsy Specimen; Blood Vessels; Brain; Brain Neoplasms; Cell surface; Cells; Cessation of life; Chimeric Proteins; Clinical Data; Data; Endothelial Cells; Evaluation; Event; Genetic; Glioblastoma; Glioma; Growth; Human; Immune; In Situ Hybridization; Injection of therapeutic agent; Integrins; Knockout Mice; Label; Malignant Glioma; Measurement; Measures; Mediating; Methods; Modeling; Mus; Nude Mice; Patients; Peptides; Platelet-Derived Growth Factor; Proto-Oncogene Proteins c-akt; Receptors, Tumor Necrosis Factor, Type II; Recurrence; Reverse Transcriptase Polymerase Chain Reaction; S-Phase Fraction; Schedule; Signal Transduction; Specificity; Stem cells; Surface; Testing; Therapeutic; Tumor Angiogenesis; Tumor Cell Invasion; Tumor Necrosis Factor Receptor; Tumor Necrosis Factor Therapy; Tumor Necrosis Factor-alpha; Tumor Volume; Tumor-Derived; Xenograft Model; alanine aminopeptidase; angiogenesis; base; density; dosage; feeding; in vivo; in vivo Model; innovation; insight; killings; laser capture microdissection; monolayer; mouse model; neovasculature; nestin protein; non-invasive monitor; novel; novel therapeutics; peripheral blood; pre-clinical; prevent; public health relevance; response; tumor; tumor growth; white matter

DESCRIPTION (provided by applicant): Anti-angiogenic approaches that eliminate the neovasculature by inducing apoptosis would represent a significant advance in the treatment of glioblastomas. Tumor necrosis factor a (TNFa) can act to induce apoptosis of cultured primary human brain microvessel endothelial cells (MvEC) through a mechanism that requires expression of the TNF-receptor 1 (TNF-R1) on the MvEC. Immunohistochemical analysis of biopsies indicates that, in most patients, the expression of TNF-R1 and TNFa is significantly higher in the glioblastoma tumor endothelial cells as compared to the normal brain endothelial cells and the levels of tumor-associated angiogenesis in tumors developed by injection and propagation of mouse malignant glioma cells in the white matter of the mouse brain is significantly higher in TNF-R1-null mice than in their wild-type counterparts. Based on these and other data, we hypothesize that the upregulated expression of TNF-R1 on brain endothelial cells associated with malignant glioma tumors is a host anti-angiogenic response to the tumor, and that TNFa therapy targeted to tumor endothelial cells will inhibit tumor angiogenesis and tumor growth. We propose to test these hypotheses by identifying the cell surface signaling events that elicit, and regulate, TNF-R1-mediated apoptosis in glioblastoma MvECs. In parallel, we will establish the feasibility of therapeutic manipulation of TNF-R1 with a TNFa fusion protein that is targeted to tumor MvECs by fusion with a peptide that binds CD13. We will use two mouse models to analyze the specificity of the effects and the magnitude of the responses in vivo: an immune competent mouse model of glioblastoma and a xenograft model based on the use of human glioblastoma stem cells. We will: (1) Establish whether TNF-R1 is preferentially expressed in the brain tumor MvEC and is colocalized with molecules that may regulate its ability to signal apoptosis, using biopsies from patients with glioblastoma and normal brain; (2) Determine whether TNF-R1 functions as an anti-angiogenic molecule in the brain in response to a malignant glioma tumor and establish whether TNF- R2 contributes to, or modulates, this effect using TNF-R1-null, TNF1-null, and TNF-R2-null mice; (3) Determine whether the activation state or expression of integrin av¿3 on the brain MvEC modulates the response of these cells to the pro-death signaling of TNF1; and (4) Test the ability of a TNFa fusion protein targeted to CD13 on tumor endothelial cells with the Cys-Asn-Gly-Arg-Cys peptide to inhibit tumor angiogenesis and tumor growth, and to promote survival, in vivo. RELEVANCE: The results should identify a novel anti-angiogenic therapy that can be used in conjunction with other therapies to more effectively eliminate malignant glioma tumors and prevent their recurrence. The studies also will provide data concerning biomarkers that may be used to predict which glioblastoma patients may benefit from this strategy and biomarkers for non-invasive monitoring of its efficacy. PUBLIC HEALTH RELEVANCE: The survival of patients with glioblastoma tumors is dismal (15-18 month median survival) despite all current therapy. We propose to test a novel therapeutic strategy to preferentially kill the endothelial cells (MvECs) in the newly formed blood vessels that feed a glioblastoma tumor by targeting tumor necrosis factor a (TNFa) to these blood vessels through a peptide (NGR) that binds CD13. CD13 is upregulated on tumor endothelial cells. Importantly, we will include innovative analyses of the molecules that may regulate the responsiveness of the MvECs. This approach would be a significant advance over the therapies currently being tested that retard the growth of the blood vessels, but do not eliminate them.

描述（由申请人提供）：通过诱导细胞凋亡来消除新血管系统的抗血管生成方法将代表胶质母细胞瘤治疗的重大进展。肿瘤坏死因子 a (TNFa) 可以通过需要在 MvEC 上表达 TNF 受体 1 (TNF-R1) 的机制来诱导培养的原代人脑微血管内皮细胞 (MvEC) 凋亡。活检的免疫组织化学分析表明，在大多数患者中，与正常脑内皮细胞相比，胶质母细胞瘤肿瘤内皮细胞中 TNF-R1 和 TNFa 的表达显着较高，并且在 TNF-R1 缺失小鼠中，通过在小鼠脑白质中注射和增殖小鼠恶性胶质瘤细胞而形成的肿瘤中肿瘤相关血管生成的水平显着较高 比其野生型对应物。基于这些和其他数据，我们假设与恶性神经胶质瘤相关的脑内皮细胞上TNF-R1的表达上调是宿主对肿瘤的抗血管生成反应，并且针对肿瘤内皮细胞的TNFa治疗将抑制肿瘤血管生成和肿瘤生长。我们建议通过识别在胶质母细胞瘤 MvEC 中引发和调节 TNF-R1 介导的细胞凋亡的细胞表面信号传导事件来检验这些假设。与此同时，我们将建立用 TNFa 融合蛋白治疗 TNF-R1 的可行性，该融合蛋白通过与结合 CD13 的肽融合来靶向肿瘤 MvEC。我们将使用两种小鼠模型来分析体内效应的特异性和反应的程度：具有免疫能力的胶质母细胞瘤小鼠模型和基于使用人类胶质母细胞瘤干细胞的异种移植模型。我们将：（1）使用胶质母细胞瘤患者和正常大脑的活检，确定 TNF-R1 是否优先在脑肿瘤 MvEC 中表达，并与可能调节其信号凋亡能力的分子共定位； (2) 使用 TNF-R1 缺失、TNF1 缺失和 TNF-R2 缺失小鼠确定 TNF-R1 是否在大脑中作为抗血管生成分子来响应恶性神经胶质瘤肿瘤，并确定 TNF-R2 是否有助于或调节这种作用； (3)确定大脑MvEC上整合素av¿3的激活状态或表达是否调节这些细胞对TNF1促死亡信号传导的反应； (4)测试具有Cys-Asn-Gly-Arg-Cys肽的靶向肿瘤内皮细胞上的CD13的TNFa融合蛋白在体内抑制肿瘤血管生成和肿瘤生长以及促进存活的能力。相关性：结果应确定一种新型抗血管生成疗法，可与其他疗法联合使用，以更有效地消除恶性神经胶质瘤并预防其复发。这些研究还将提供有关可用于预测哪些胶质母细胞瘤患者可能受益于该策略的生物标志物的数据，以及用于非侵入性监测其疗效的生​​物标志物的数据。 公共卫生相关性：尽管目前采用了所有治疗方法，但胶质母细胞瘤患者的生存率仍然很低（中位生存期为 15-18 个月）。我们建议测试一种新的治疗策略，通过结合 CD13 的肽 (NGR) 将肿瘤坏死因子 a (TNFa) 靶向这些血管，从而优先杀死为胶质母细胞瘤肿瘤供血的新形成血管中的内皮细胞 (MvEC)。 CD13 在肿瘤内皮细胞上表达上调。重要的是，我们将包括对可能调节 MvEC 反应性的分子的创新分析。与目前正在测试的延缓血管生长但不能消除血管的疗法相比，这种方法将是一个重大进步。