A2CPS ExRNA Component

细胞外RNA组分 总结 细胞外RNA（exRNA）已被发现在所有测试的人类生物流体，有越来越多的证据表明， 它们在疼痛介导和调节中发挥作用，并且可以作为生物标志物用于识别那些在 急性疼痛发作后转变为慢性疼痛的风险。这个项目是exRNA组件的 急性至慢性疼痛特征（A2 CPS）项目的组学数据生成中心（ODGC）。在这 根据该计划，临床中心将从两个纵向研究中心招募并收集临床数据和生物流体样本。 每组1800名受试者。将在急性疼痛发作后0、3和6个月采集生物流体样本， 由特定的外科手术或特定的肌肉骨骼创伤组成。这些样本将用于 生成多组学数据，以验证40个主要结果生物标志物，这些生物标志物指示对 慢性疼痛的发展，以及确定新的候选生物标志物。对于拟议的exRNA 目标1的组成部分将在第一年执行，它将涉及与联合国其他组成部分的密切合作。 A2 CPS计划，以建立最终研究设计和方案。所有A2 CPS项目的研究人员 将共同努力建立40个主要结果生物标志物。ODGC和临床中心研究者 将共同决定具体的样本类型和采集/处理/储存方法。ODG和 数据集成资源中心/数据协调组件（DIRC/DCC）调查人员将建立 元数据和数据标准以及向DCC提交元数据和数据的工作流程。exRNA ODGC的组成部分和行政核心将建立一个LIMS，用于样本和数据跟踪， 记录元数据。exRNA组件和DIRC/数据集成和分析组件（DIAC）将 建立qRT-PCR和小型RNAseq数据分析管道。目标2和3将跨越第2-4年，目标2 研究重点是从约11，000份生物流体样品中分离和qRT-PCR以及exRNA的小RNAseq分析， 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。本组件的主要目标 是生成和提交高质量的exRNA qRT-PCR和小RNAseq数据，用于验证预 选择候选exRNA生物标志物并发现新型exRNA生物标志物。exRNA组分 研究者还期望参与DIRC/DIAC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、心理社会和/或成像）组成的特征 生物标志物），指示对慢性疼痛的易感性/弹性，其可用于开发个性化的 预防和治疗慢性疼痛的策略。该拟定exRNA组件的人员 通过参与，获得了成功执行该项目所需的技能和知识， 细胞外RNA通讯联盟，包括细胞外囊泡的高通量工作流程- 特异性和总exRNA分离以及小RNAseq文库制备和exRNA数据分析管道。

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.