Modified RNA tools and diagnostics for drug abuse

改良 RNA 工具和药物滥用诊断

基本信息

批准号：
8841583
负责人：
PAUL F AGRIS
金额：
$ 22.5万
依托单位：
ATHGHIN BIOTECHNOLOGY, INC.
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-09-01 至 2016-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8841583
关键词：
Adopted Affect Anterior Biological Biological Assay Biological Markers Biological Process Brain Cannabinoids Cell physiology Cells Cerebral cortex Chemical Exposure Chemicals Chemistry Chronic Communities Complex Computer software Cues Detection Development Diagnostic Disease Drug Exposure Drug abuse Epigenetic Process Functional RNA Gene Expression Gene Expression Regulation Goals Health High Pressure Liquid Chromatography Human Investigation Legal Liquid substance Mass Spectrum Analysis Measures Mental disorders Messenger RNA Metabolic Methods Methylation Modification Molecular Nervous system structure Neuroglia Neurons Nicotine Nucleosides Pathology Pattern Pharmaceutical Preparations Play Prosencephalon RNA RNA Degradation RNA Stability Reporting Research Resolution Role Sample Size Specificity Speed Staging Structure Substance abuse problem Substance of Abuse Technical Expertise Technology Testing abstracting analytical method base biological systems brain cell computerized data processing drug of abuse induced pluripotent stem cell insight instrumentation liquid chromatography mass spectrometry nervous system disorder novel public health relevance tool

项目摘要

DESCRIPTION (provided by applicant): Project Summary/Abstract RNA modifications are numerous, and range from simple methylation to complex changes such as addition of aminoacylcarbamoyl moieties. These modifications and, the levels at which are present, are emerging as important regulators of biological processes, and perturbations such as chemical or drug exposure can alter the patterns of specific modifications on RNA sub-types. Moreover, given their variety, specificity and their relationship to function and structure stability, RNA modifications are potentially valuable biomarkers of disease and treatment progression, especially given that modified nucleosides are relatively stable even after RNA degradation, and can be released from cells into bodily fluids. The long term objective of this study is to facilitae research into RNA modifications and their impact on health and disease. The immediate hurdle, and the goal of this application, is to develop extremely sensitive analytical methods for detection and quantification of common and novel RNA modifications that can be readily disseminated to the RNA research community. The test case biological system is based on human induced pluripotent stem cell derived nervous system cells that will be exposed to two different drugs of abuse, nicotine and cannabinoids. Aim 1. We will use RNA modified nucleoside standards to develop highly sensitive and robust method of analysis. The ultimate tool we will be developing is a product resultant from the combination of two technologies, UHPLC and MS that will be optimized for high resolution separation and highly sensitive detection of RNA modifications. The tools developed will be tested on differentiated anterior cerebral cortex neurons and glia cells and RNA modifications will be characterized and abundances quantified during the different stages of maturation. Aim 2. Tools developed during aim 1 will be further adapted to detect specific RNA modifications that may be playing a significant role on differentiated forebrain cells, as a result from treatment with drug of abuse. During this aim we will specifically study forebrain cells that will be exposed to different nicotie and cannabinoids concentrations and compared to vehicle controls. The results of the methods we are proposing will be in the form of highly sensitive and accurate tools for the analysis of a wide range of chemistries and abundances of RNA modifications present in regulatory RNAs. The ultimately goal is to develop highly sensitive RNA modifications-based biomarkers discovery tools that requires minimal level of technical expertise as a way to disseminate and accelerate the field of RNA modifications with regards to gain further insight into mechanisms of disease.

描述（由申请人提供）：项目摘要/摘要 RNA 修饰有很多，范围从简单的甲基化到复杂的变化，例如添加氨酰基氨基甲酰基部分。这些修饰及其存在的水平正在成为生物过程的重要调节因子，而化学或药物暴露等扰动可以改变 RNA 亚型的特定修饰模式。此外，鉴于RNA修饰的多样性、特异性及其与功能和结构稳定性的关系，RNA修饰是疾病和治疗进展的潜在有价值的生物标志物，特别是考虑到修饰的核苷即使在RNA降解后也相对稳定，并且可以从细胞释放到体液中。这项研究的长期目标是促进 RNA 修饰及其对健康和疾病影响的研究。该应用的直接障碍和目标是开发极其灵敏的分析方法，用于检测和定量常见和新颖的 RNA 修饰，并且可以轻松传播到 RNA 研究界。测试案例生物系统基于人类诱导多能干细胞衍生的神经系统细胞，这些细胞将暴露于两种不同的滥用药物：尼古丁和大麻素。目标 1. 我们将使用 RNA 修饰的核苷标准品来开发高灵敏度和稳健的分析方法。我们将开发的最终工具是 UHPLC 和 MS 两种技术相结合的产品，该产品将针对高分辨率分离和 RNA 修饰的高灵敏度检测进行优化。开发的工具将在分化的前大脑皮层神经元和神经胶质细胞上进行测试，并且将在成熟的不同阶段对 RNA 修饰进行表征和丰度量化。目标 2. 在目标 1 期间开发的工具将进一步适应于检测特定的 RNA 修饰，这些修饰可能在滥用药物治疗导致的分化前脑细胞中发挥重要作用。在此目标期间，我们将专门研究将暴露于不同尼古丁和大麻素浓度的前脑细胞，并与载体对照进行比较。我们提出的方法的结果将以高度灵敏和准确的工具的形式出现，用于分析调节 RNA 中存在的各种化学成分和丰度的 RNA 修饰。最终目标是开发高度敏感的基于 RNA 修饰的生物标志物发现工具，该工具需要最低水平的技术专业知识，作为传播和加速 RNA 修饰领域的一种方式，以进一步了解疾病机制。