In situ assay imaging nuclear RNA exosome activity for cancer studies

用于癌症研究的核RNA外泌体活性原位测定成像 摘要 该项目的目标是初步开发和演示一种新的分子技术， 提供了高度新颖的测量和靶向能力，可能会改变癌症研究。这 创新的方法将能够对RNA周转的分子机制进行新型评估， for cancer癌症biology生物学.该项目将引入第一个能够标记RNA降解的原位技术 核RNA外泌体的活性。RNA外泌体（不要与不相关的囊泡外泌体混淆）是 控制细胞中RNA代谢的主要酶复合物。它是生命所必需的。其基本 其功能是保持细胞处于增殖状态。过度活跃的外泌体复合物导致更高的 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 抗癌治疗的方法。核RNA外泌体活性在评估肿瘤细胞应激和细胞凋亡中至关重要。 死亡倾向，以及评估癌症对治疗的反应。 尽管在固定的细胞和组织中原位标记RNA外泌体活性的测定具有很高的实用性， 目前还没有这样的成像技术。该过程目前正在研究通过使用散装 在异质组织样品中具有有限价值的生物化学方法。 在这个项目中，我们将克服这一限制，并将开发第一个用于标记核活性的检测方法， 固定组织切片形式的RNA外泌体。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.首次建立在固定组织切片上特异性标记核RNA外泌体活性的方法 格式.该方法将允许通过使用创新的加帽的 杂交RNA探针。 2.为了测试和验证新开发的原位标记技术的核心功能， 来自具有活化的和正常的核RNA外泌体活性的模型的组织切片，包括胶质母细胞瘤。 优化新方法的特异性、灵敏度，保证检测的可靠性。

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.