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: 8223277
• 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/8223277
• 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（由适用提供）：通过诱导的凋亡消除新生血管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发性人脑微血管内皮细胞（MVEC）凋亡。对活检的免疫组织化学分析表明，与正常的脑内皮细胞相比，在大多数患者中，在胶质母细胞瘤肿瘤内皮细胞中，TNF-R1和TNFA的表达明显更高比在野生型的对应物中。基于这些和其他数据，我们假设TNF-R1在与恶性神经胶质瘤肿瘤相关的脑内皮细胞上的更新表达是对肿瘤的宿主抗血管生成反应，而针对肿瘤内皮细胞的TNFA治疗将抑制肿瘤内皮细胞抑制肿瘤血管生成和肿瘤生长。我们建议通过鉴定引起和调节胶质母细胞瘤MVEC中TNF-R1介导的凋亡的细胞表面信号传导事件来检验这些假设。同时，我们将与TNF-R1对TNFA融合蛋白进行治疗操作的可行性，该蛋白通过与结合CD13的肽融合靶向肿瘤MVEC。我们将使用两个小鼠模型来分析体内效应的特异性和反应的大小：胶质母细胞瘤的免疫能力小鼠模型和基于人类胶质母细胞瘤干细胞的使用的异种移植模型。我们将：（1）确定TNF-R1是否优选在脑肿瘤MVEC中表达，并与可能调节其信号凋亡能力的分子共定位，使用胶质母细胞瘤和正常脑患者的活检； （2）确定TNF-R1是否在响应恶性神经胶质瘤肿瘤的大脑中充当抗血管生成分子，并确定TNF-R2是使用TNF-R1-NULL，TNF1-NULL，TNF1-NULL和TNF-R2-NULL小鼠有助于或调节这种作用的； （3）确定TNF-R1是否在响应恶性神经胶质瘤肿瘤的大脑中充当抗血管生成分子，并确定使用TNF-R1-NULL，TNF1-NULL，TNF1-NULL和TNF-R2-NULL小鼠对TNF-R2是否有助于或调节这种作用； （3）确定整联蛋白AV¿3在脑MVEC上的激活状态或表达是否会调节这些细胞对TNF1的促进死亡信号的反应； （4）测试具有Cys-Asn-Gly-Arg-Cys肽抑制肿瘤血管生成和肿瘤生长的TNFA融合蛋白针对CD13的TNFA融合蛋白的能力，并促进生存。相关性：结果应确定一种新型的抗血管生成疗法，可以与其他疗法结合使用，以更有效地消除恶性神经胶质瘤肿瘤并防止其复发。这些研究还将提供有关生物标志物的数据，这些数据可用于预测哪些胶质母细胞瘤患者可能会受益于该策略和生物标志物，以无创监测其有效性。 公共卫生相关性：胶质母细胞瘤肿瘤患者的存活率令人沮丧（15-18个月的中位数生存期）希望所有当前的疗法。我们建议测试一种新型的治疗策略，以优先杀死新形成的血管中的内皮细胞（MVEC），该血管通过将肿瘤坏死因子A（TNFA）靶向通过结合CD13的肽（NGR）靶向这些血管，以靶向肿瘤坏死因子A（TNFA）。 CD13对肿瘤内皮细胞进行了更新。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。对于当前正在测试的疗法，这种方法将是一个重大进步，该疗法阻碍了血管的生长，但不会消除它们。