In situ detection of apoptosis in tumor endothelial cells

描述：(申请人提供) 靶向肿瘤血管的药物具有巨大的抗癌潜力 治疗方法，因为它们似乎不会引起耐药性。然而， 这些药物具有细胞抑制作用，而不是细胞毒性，这就形成了一套独特的 药物疗效评价中存在的问题，因为经典的药效评价方法 肿瘤的“反应”可能是无效的。尤其重要的是， 确定它们的生物学作用机制并开发新的方法来 在初诊患者的组织标本中检测这些影响。为此，我们 已经开发出一种技术，可以检测濒临死亡的肿瘤 内皮细胞，我们的初步结果强烈表明，抗血管生成 药物可诱导肿瘤内皮细胞的凋亡。整体而言 本研究的目的是确定内皮细胞是否发生了凋亡 是抗血管生成药物治疗肿瘤的敏感标记物或疗效 可以区分对这些药物有反应的患者的治疗方法 谁不会呢。为此，我们建议：(1)界定特定的角色 “存活”通路在维持体外内皮细胞活性中的作用。 细胞将暴露于生长因子停用或血管内皮生长因子受体 拮抗剂，以及对先前涉及细胞内的信号通路的影响 将评估存活率(即AKT)。我们还将从这些细胞中分离出mRNAs 细胞，并分析细胞凋亡相关基因表达的变化 停用血管内皮细胞生长因子。(2)确定内皮细胞凋亡在血管内皮细胞损伤中的作用 原位肿瘤模型。裸鼠携带人胰腺、结肠或 前列腺癌将用研究药物治疗，并对 用CD31/TUNEL染色检测肿瘤内皮细胞的凋亡率。(3) 血管内皮细胞凋亡在抗血管生成作用中的作用 患者的治疗方法。血管内皮细胞的凋亡水平 接受抗血管生成药物治疗的患者将与肿瘤相关 血流变化及放射学测量的反应。这些研究 将使我们能够快速确定肿瘤内皮细胞的凋亡 可作为临床疗效的替代物。 一类新型抗癌药物。

Description: (provided by applicant) Agents that target tumor vasculature have tremendous potential as anti-cancer therapies because they do not appear to induce drug resistance. However, these agents are cytostatic rather than cytotoxic, which poses a unique set of problems for the evaluation of drug efficacy because classical measures of tumor "response" may not be valid. It will be especially important to identify their biological mechanisms of action and to develop new methods to detect these effects in in primary patient tissue specimens. To this end, we have developed a technique that allows for the detection of dying tumor endothelial cells, and our preliminary results strongly suggest that antiangiogenic agents induce apoptosis in tumor endothelial cells. The overall goal of the present studies is to determine whether endothelial cell apoptosis is a sensitive marker or efficacy in tumors treated with anti-angiogenic therapies that can distinguish patients who respond to these drugs from those who do not. To this end, we propose to: (1) Define the role of specific "survival" pathways in the maintenance of endothelial cell viability in vitro. Cells will be exposed to growth factor withdrawal or VEGF receptor antagonists, and effects on signaling pathways previously implicated in cell survival (i.e. AKT) will be evaluated. We will also isolate mRNA from these cells and analyze changes in apoptosis-associated gene expression following VEGF withdrawal. (2) Determine the role of endothelial cell apoptosis in orthotopic tumor models. Nude mice bearing human pancreatic, colon, or prostate tumors will be treated with investigational agents, and effects on tumor endothelial cell apoptosis will be measured by CD31/TUNEL staining. (3) Characterize the role of endothelial cell apoptosis in the effects of antiangiogenic therapies in patients. Levels of endothelial cell apoptosis in patients treated with anti-angiogenic agents will be correlated with tumor blood flow changes and radiographic measurements of response. These studies will allow us to rapidly determine whether tumor endothelial cell apoptosis can be used as a surrogate for clinical response in patients treated with this class of novel anti-cancer agents.