A2CPS ExRNA Component

ExTRACELLAR RNA组件 摘要 在所有被测试的人体体液中都发现了细胞外RNA(ExRNAs)，而且有越来越多的证据表明 它们在疼痛调节和调节中发挥作用，并可以作为识别疼痛的生物标志物。 在急性疼痛发作后过渡到慢性疼痛的风险。此项目是ExRNA组件 急性到慢性疼痛信号(A2CPS)计划的组学数据生成中心(ODGC)。在这 该计划，临床中心将招募和收集临床数据和生物液样本从纵向 每组1800名受试者。生物体液样本将在急性疼痛发作后0、3和6个月采集， 由特定的外科手术或特定的肌肉骨骼创伤组成。这些样品将被用来 生成多组数据以验证40个主要结果生物标记物，这些生物标记物表明对 慢性疼痛的发展，以及确定新的候选生物标记物。对于建议的exRNA 将在第一年执行的AIM 1组件将与其他组件密切合作 A2CPS计划建立最终的研究设计和方案。所有A2CPS计划调查员 将共同努力建立40个主要结果生物标记物。ODGC和临床中心调查人员 将共同决定具体的样品类型(S)和采集/加工/存储方法。ODGC和 数据集成资源中心/数据协调部门(DIRC/DCC)调查员将建立 元数据和数据标准以及向发改委提交元数据和数据的工作流程。外源RNA 该司的构成部分和行政核心将建立一个用于样本和数据跟踪的LIMS，以及 记录元数据。ExRNA组件和DIRC/数据集成和分析组件将 建立qRT-PCR和Small RNAseq数据分析管道。目标2和目标3将跨越2-4年，目标2 重点是从约11,000个生物液样本中分离、qRT-PCR和小RNAseq图谱 将由临床中心收集。目标3将包括将元数据和数据提交给 DIRC/DCC、数据质量控制以及数据分析和解释。此组件的主要目标是 是否生成并提交高质量的exRNA qRT-PCR和小RNAseq数据，以验证Pre- 筛选候选exRNA生物标记物和发现新的exRNA生物标记物。外源RNA组分 调查人员还预计将与DIRC/DIAC一起参与旨在发展疼痛的综合分析 由多种生物标志物类型(包括分子、临床、心理和/或成像)组成的签名 生物标记物)指示对慢性疼痛的敏感性/弹性，可用于发展个性化 预防和治疗慢性疼痛的策略。提议的exRNA组件的工作人员 我已通过参与该项目获得了成功执行该项目所需的技能和知识 细胞外RNA通信联盟，包括细胞外小泡的高通量工作流- 特异性和全量exRNA的分离及小RNAseq文库的制备和exRNA数据分析的流水线。

EXTRACELLAR RNA COMPONENT SUMMARY Extracellular RNAs (exRNAs) have been found in all tested human biofluids, and there is increasing evidence that they play a role in pain mediation and modulation, and can serve as biomarkers for identification of those at risk for transitioning to chronic pain after an acute pain episode. This project is the exRNA Component of the Omics Data Generation Center (ODGC) for the Acute to Chronic Pain Signatures (A2CPS) Program. In this Program, the Clinical Centers will recruit and collect clinical data and biofluid samples from two longitudinal cohorts of 1800 subjects each. Biofluid samples will be collected 0, 3, and 6 months after an acute pain episode, consisting of a specific surgical procedure or a specific musculoskeletal trauma. These samples will be used to generate multi-omic data to validate 40 primary outcome biomarkers indicating susceptibility or resilience to development of chronic pain, as well as to identify new candidate biomarkers. For the proposed exRNA Component, Aim 1, which will be executed in Year 1, will involve close collaboration with other components of the A2CPS Program to establish the final study design and protocols. All of the A2CPS Program investigators will work together to establish the 40 primary outcome biomarkers. The ODGC and Clinical Center investigators will jointly decide on the specific sample type(s) and collection/processing/storage methods. The ODGC and Data integration Resource Center/Data Coordination Component (DIRC/DCC) investigators will establish Metadata and Data Standards and a workflow for submission of metadata and data to the DCC. The exRNA Component and the Administrative Core of the ODGC will establish a LIMS for sample and data tracking, and recording of metadata. The exRNA Component and DIRC/Data Integration and Analysis Component (DIAC) will establish qRT-PCR and small RNAseq data analysis pipelines. Aims 2 and 3 will span Years 2-4, with Aim 2 focused on isolation and qRT-PCR and small RNAseq profiling of exRNA from the ~11,000 biofluid samples that will be collected by the Clinical Centers. Aim 3 will encompass submission of metadata and data to the DIRC/DCC, quality control of the data, and data analysis and interpretation. The primary goal of this Component is generation and submission of high-quality exRNA qRT-PCR and small RNAseq data for validation of pre- selected candidate exRNA biomarkers and discovery of novel exRNA biomarkers. The exRNA component investigators also anticipate participating in integrative analyses with the DIRC/DIAC aimed at developing pain signatures comprised of multiple biomarker types (including molecular, clinical, psychosocial, and/or imaging biomarkers) indicating susceptibility/resilience to chronic pain, which can be used to develop personalized strategies for prevention and treatment of chronic pain. The personnel for this proposed exRNA Component have acquired the necessary skills and knowledge to successfully execute this project from participation in the Extracellular RNA Communication Consortium, including high-throughput workflows for extracellular vesicle- specific and total exRNA isolation and small RNAseq library preparation and exRNA data analysis pipelines.