TUMOR-DERIVED VASCULAR PERMEABILITY FACTOR & BRAIN EDEMA

肿瘤源性血管通透性因子

• 批准号: 2095908
• 负责人: GREGORY R CRISCUOLO
• 金额: $ 9.59万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-04-01 至 1996-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2095908
• 关键词: antiinflammatory agents; benzodiazepines; blood brain barrier; brain edema; brain metabolism; brain neoplasms; calcium channel blockers; calcium flux; cardiovascular disorder chemotherapy; corticosteroids; dexamethasone; dihydropyridines; dyes; fluorescence spectrometry; glioma; hormone therapy; human tissue; laboratory rat; phenylalkylamine; radiotracer; retina; tissue /cell culture; vascular endothelium permeability

The long-term objective of this research program is to advance the current knowledge of blood-brain barrier disruption in the setting of tumor-associated brain edema. This will be accomplished by studying a vascular permeability factor (VPF), that is expressed by malignant glial tumors grown in tissue culture. In addition to promoting microvascular permeability, partially purified VPF is known to induce transient changes in endothelial cytosolic calcium. Of particular interest is how the VPF-induced intracellular calcium ion flux, alterations in the F-actin content, and changes in endothelial cytoarchitecture, may relate to impaired blood-brain barrier integrity in the setting of intracerebral tumor. Furthermore, the mechanism of glucocorticosteroid-induced inhibition of VPF activity, and its relevance to the known clinical efficacy of dexamethasone in the setting of neoplastic brain edema will be studied. This research seeks to supplement the current understanding of neoplastic blood-brain barrier disruption in hopes of proposing novel and more effective therapeutic alternatives. Specifically, to test the hypothesis that: VPF plays an integral role in the genesis of neoplastic brain edema. The following specific aims have been devised to support or test this hypothesis. To determine if: (1) VPF is capable of inducing cytosolic calcium changes in cultured brain and retinal endothelial cells, (2) VPF is capable of altering the cytoarchitecture of brain and retinal endothelial cells grown in monolayer cultures, (3) VPF is capable of altering the physiological barrier integrity of brain and retinal endothelial cells grown in monolayer cultures, (4) VPF activity may be inhibited in vitro and in vivo is; and to propose that effective inhibitory agents (VPF antagonists) may be applicable to the treatment of tumor-induced vasogenic brain edema. The methodology will include standard tissue culture techniques, use of a biological assay for quantifying VPF induced microvascular permeability, determination of endothelial cytosolic calcium changes in response to VPF exposure using a fluorescent intracellular calcium ion probe (fura-2/AM), monitoring endothelial cells for evidence of VPF-induced cytoarchitectural changes. by assaying for F-actin and by using electron microscopy, and using radiolabeled albumin to study the barrier integrity of endothelial cells grown in monolayers. The health-relatedness of this project derives from the substantial morbidity associated with peritumoral brain edema. Improvements in therapeutic intervention will result from a more complete understanding of this process. Ultimately, effective treatment of neoplastic brain edema will reduce the risk of surgically-induced neurological deficits, and improve patient tolerance of adjunctive radiotherapy, chemotherapy, and immunotherapy.

该研究计划的长期目标是推进当前的 血脑屏障破坏的知识 肿瘤相关脑水肿。 这将通过研究来完成 血管通透性因子（VPF），由恶性胶质细胞表达 在组织培养中生长的肿瘤。 除了促进微血管 渗透性，部分纯化的 VPF 已知会引起瞬时变化 内皮细胞质钙。 特别令人感兴趣的是如何 VPF 诱导的细胞内钙离子通量，F-肌动蛋白的变化 含量和内皮细胞结构的变化可能与 脑内出血时血脑屏障完整性受损 瘤。 此外，糖皮质激素诱导的机制 VPF 活性的抑制及其与已知临床的相关性 地塞米松治疗肿瘤性脑水肿的疗效 研究过。 本研究旨在补充目前的理解 肿瘤性血脑屏障破坏，希望提出新的和 更有效的治疗替代方案。 具体来说，要测试 假设：VPF 在肿瘤的发生中起着不可或缺的作用 脑水肿。 制定了以下具体目标来支持或 检验这个假设。 确定： (1) VPF 是否能够诱导 培养的大脑和视网膜内皮细胞中胞质钙的变化， (2) VPF能够改变大脑和视网膜的细胞结构 在单层培养物中生长的内皮细胞，(3) VPF 能够 改变大脑和视网膜的生理屏障完整性 单层培养物中生长的内皮细胞，(4) VPF 活性可能是 体外和体内均受到抑制；并提出有效的抑制 药物（VPF拮抗剂）可能适用于治疗 肿瘤引起的血管源性脑水肿。 该方法将包括标准 组织培养技术，使用生物测定来量化 VPF 诱导微血管通透性、内皮细胞质测定 使用荧光灯响应 VPF 暴露后钙的变化 细胞内钙离子探针（fura-2/AM），监测内皮细胞 寻找 VPF 诱导的细胞结构变化的证据。通过测定 F-肌动蛋白并通过使用电子显微镜和放射性标记的白蛋白来 研究单层生长的内皮细胞的屏障完整性。 这 该项目的健康相关性源于大量的发病率 与瘤周脑水肿有关。 治疗方面的改进 更全面地了解这一点将导致干预 过程。 最终，有效治疗肿瘤性脑水肿将 降低手术引起的神经功能缺损的风险，并改善 患者对辅助放疗、化疗的耐受性 免疫疗法。