Apoptosis driven by elastase inhibitor: new approach to detection and study.

描述（由申请人提供）：已发现导致癌细胞凋亡的多种途径。为了了解在每种特定情况下控制细胞消除的机制，有必要使用可以区分不同凋亡途径的探针。最近鉴定的凋亡程序之一是弹性蛋白酶促凋亡。然而，这种特殊的途径仍然缺乏特定的工具，其可视化的组织切片格式，这是最相关的病理学研究。单核细胞中性粒细胞弹性蛋白酶抑制剂（MNEI）是该途径的主要成员，负责其激活。它通过自我转化为活性核酸酶来发挥作用，并能迅速处理肿瘤细胞。促进脑肿瘤中这种独特转变的条件目前尚未完全了解。在很大程度上，这是因为在组织切片和活细胞培养物中缺乏用于选择性标记MNEI驱动的细胞凋亡的特异性工具。在缺乏特定的原位方法，这种凋亡途径，它是目前研究通过使用大量的生化方法，在异质性病理学样本的价值有限。在这个项目中，我们将克服这一障碍，并将开发新的成像技术。我们将应用新的探针来可视化胶质母细胞瘤（GBM）中MNEI驱动的细胞凋亡。具体目标：1.建立第一个选择性检测组织切片中弹性蛋白酶抑制剂驱动的凋亡途径的方法。该方法将标记该途径的关键事件，并可视化单核细胞中性粒细胞弹性蛋白酶抑制剂（MNEI）转化为活性DNA酶后的特征性DNA切割活性。在胶质母细胞瘤切片中测试这种新开发的成像工具。2.建立一种新的选择性标记弹性蛋白酶激活的细胞凋亡途径的方法。特异于MNEI途径的新荧光传感器仅在检测到该凋亡途径的特异性标记物后才会产生荧光。该项目将为一般的细胞凋亡研究，特别是癌症研究引入使能技术。它们在胶质母细胞瘤中的应用将为这些肿瘤的未来临床和研究调查以及有效治疗干预的发展提供有用的信息。

DESCRIPTION (provided by applicant): Multiple pathways leading to apoptosis of cancer cells have been discovered. To understand the mechanisms controlling cell elimination in each specific situation, it is necessary to have probes which can distinguish between different apoptotic routes. One of the most recently identified apoptotic programs is the elastase inhibitor-driven apoptosis. However, this particular pathway still lacks specific tools for its visualizationin the tissue section format, which is most relevant for pathology studies. Monocyte neutrophil elastase inhibitor (MNEI) is the main member of this pathway responsible for its activation. It acts by self-converting into an active nuclease and can rapidly dispose of tumor cells. The conditions promoting such a unique transition in brain tumors are not fully understood at this time. To a large extent this is because of the absence of specific tools for selective labeling of MNEI-driven apoptosis in tissue sections and in live cell cultures. In the absence of specific in situ methods for this apoptotic pathway, it is currently studied by using the bulk biochemical approaches which have limited value in heterogeneous pathology samples. In this project we will overcome this obstacle and will develop new enabling imaging technologies. We will apply the new probes to visualize MNEI-driven apoptosis in glioblastoma (GBM). Specific aims: 1. To develop the first methodology for selective detection of the elastase inhibitor- driven apoptotic pathway in tissue sections. The approach will label the key event of this pathway and visualize the characteristic DNA cleavage activity of monocyte neutrophil elastase inhibitor (MNEI) after its transformation into an active DNase. To test this newly developed imaging tool in glioblastoma sections. 2. To develop the new methodology for selective labeling of the elastase inhibitor-driven apoptotic pathway in live cell cultures. The new fluorescent sensors specific for MNEI pathway will produce fluorescence only after they detect a specific marker of this apoptotic route. The project will introduce enabling technologies for apoptosis research in general, and for cancer studies in particular. Their application to glioblastoma will provide information useful for future clinical and research investigations of these tumors, and for the development of effective therapeutic interventions.