In situ assay imaging nuclear RNA exosome activity for cancer studies

肿瘤研究中核糖核酸外切体活性的原位检测 摘要 该项目的目标是初步开发和示范一种新的分子技术 提供高度新颖的测量和目标定位能力，可能对癌症研究产生革命性影响。这 创新的方法将使对RNA周转的分子机制的新型评估成为可能，这是至关重要的 癌症生物学的研究。该项目将引入第一项能够标记降解的RNA的原位技术 核糖核酸外切体的活性。RNA外切体(不要与无关的囊泡外切体混淆)是 控制细胞内RNA代谢的主要酶复合体。它是生活的必需品。它的基本原理 其功能是使细胞保持在增殖状态。过度活跃的外切体复合体会导致更高的 细胞增殖，并与癌症的发展和进展有关。它也是一个关键的分子靶点。 抗癌疗法。核糖核糖核酸外切体活性在肿瘤细胞应激和细胞评估中的关键作用 死亡倾向，以及评估癌症对治疗的反应。 尽管在固定细胞和组织中原位标记RNA外切体活性的方法具有很高的实用性 节段，目前还没有这样的成像技术。目前对该过程的研究是通过使用Bulk 生物化学方法在异质组织样本中的价值有限。 在这个项目中，我们将克服这一限制，并将开发第一个标记核活性的方法。 固定组织切片格式的RNA外切体。该项目将展示 新的分子成像技术在癌症研究中具有广泛的适用性。 该提案的具体目标是： 1.首次建立了固定组织切片中核糖核酸外切体活性的特异性标记方法 格式化。该方法将允许通过使用创新的Caped 混合RNA探针。 2.测试和验证新开发的原位标记技术的核心功能能力 包括胶质母细胞瘤在内的核糖核酸外切体活性正常的组织切片。 优化新方法的特异度、灵敏度，保证检测的稳健可靠性。

In situ assay imaging nuclear RNA exosome activity for cancer studies Abstract The goal of this project is the initial development and demonstration of a new molecular technology which offers highly novel measurement and targeting capabilities potentially transformative for cancer research. This innovative approach will enable a new type assessment of molecular mechanisms of RNA turnover, essential for cancer biology. The project will introduce the first in situ technology capable of labeling the RNA degrading activity of nuclear RNA exosome. RNA exosome (not to be confused with the unrelated vesicular exosomes) is the major enzymatic complex controlling RNA metabolism in cells. It is essential for life. Its fundamental function is to keep cells in the proliferating state. An overactive exosome complex leads to higher rates of cellular proliferation and is implicated in cancer development and progression. It is also a key molecular target of anticancer therapies. Nuclear RNA exosome activity is critical in assessments of tumor cell stress and cell death propensity, and in evaluating cancer response to therapies. In spite of the high utility of an assay labeling RNA exosome activity in situ, in fixed cells and tissue sections, presently there is no such imaging technology. The process is currently studied by using bulk biochemical approaches which have limited value in heterogeneous tissue samples. In this project we will overcome this limitation and will develop the first assay for labeling activity of nuclear RNA exosome in the fixed tissue section format. The project will demonstrate the core functional capabilities of the new molecular imaging technology with wide applicability in cancer studies. Specific Aims of the proposal are: 1. To develop the first approach for specific labeling of nuclear RNA exosome activity in the fixed tissue section format. The approach will permit visualization of nuclear exosome activity by using the innovative capped hybrid RNA probe. 2. To test and validate the core functional capabilities of the newly developed in situ labeling technique in tissue sections from models with activated and normal nuclear RNA exosome activity including glioblastoma. To optimize the new method’s specificity, sensitivity and assure the robust reliability of detection.