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

基本信息

批准号：
10682455
负责人：
VLADIMIR V DIDENKO
金额：
$ 17.05万
依托单位：
BAYLOR COLLEGE OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-09-10 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10682455
关键词：
Acceleration Antineoplastic Agents Biochemical Biological Assay Biomedical Technology Cancer Biology Cell Death Cell Proliferation Cell division Cells Cellular Stress Chronic Myeloid Leukemia Complex Confusion Core Protein Detection Development Fluorouracil Glioblastoma Goals Heart Diseases Histologic Hybrids Image Imaging technology In Situ Individual Label Life Literature Lung Malignant Neoplasms Malignant neoplasm of prostate Measurement Measures Methods Modeling Molecular Molecular Target Nuclear Nuclear RNA Pathology Pathway interactions Performance Process Proliferating RNA RNA Decay RNA Probes RNA marker RNA metabolism Research Ribonucleases S-Phase Fraction Sampling Shapes Solid Neoplasm Specificity Stroke Techniques Technology Testing Time Tissue Sample Tissues Visualization Work anticancer research assay development biomedical imaging cancer therapy cell fixing exosome improved in situ imaging innovation innovative technologies melanoma molecular imaging neoplastic cell novel novel anticancer drug proliferation potential sample archive tissue fixing treatment response tumor

项目摘要

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.

原位成像测定核 RNA 外泌体活性用于癌症研究抽象的该项目的目标是初步开发和示范一种新的分子技术，该技术提供高度新颖的测量和靶向功能，可能对癌症研究产生变革。创新方法将能够对 RNA 周转的分子机制进行新型评估，这是至关重要的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）是控制细胞内 RNA 代谢的主要酶复合物，是生命的基础。功能是保持细胞处于增殖状态，过度活跃的外泌体复合物会导致更高的增殖率。细胞增殖并与癌症的发生和进展有关。它也是一个关键的分子靶点。核 RNA 外泌体活性对于评估肿瘤细胞应激和细胞至关重要。死亡倾向，以及评估癌症对治疗的反应。尽管原位标记 RNA 外泌体活性的测定在固定细胞和组织中具有很高的实用性目前还没有这样的成像技术，目前正在研究使用块体。生化方法在异质组织样本中价值有限。在这个项目中，我们将克服这一限制，并将开发第一个用于核标记活性的测定方法。该项目将展示固定组织切片形式的 RNA 外泌体的核心功能。新型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. 开发第一种在固定组织切片中特异性标记核RNA外泌体活性的方法该方法将允许通过使用创新的上限来可视化核外泌体活动。杂交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来自具有激活和正常核 RNA 外泌体活性（包括胶质母细胞瘤）的模型的组织切片。优化新方法的特异性、灵敏度，保证检测的稳健可靠性。