Defining A Comprehensive Reference Profile of Circulating Human Extracellular RNA

描述（由申请人提供）： 我们的目标是使用全面的RNA测序在来自健康人的12种不同体液样品中生成短、长和环状非编码调节exRNA（包括环境来源的exRNA）的参考谱。我们组建了一支多学科团队，拥有临床研究，基因组学技术，生物信息学和统计学方面的专业知识 来实现这一目标。该提案的一个主要优势是，我们受益于与其他研究人员的密切互动，这些研究人员参与了由共同基金细胞外RNA通讯计划支持的两个基于UCSF的U19项目。另一个主要优势是，我们将利用从参加NIH资助的队列研究的不同种族的女性和男性儿童和成年人中收集的大量精心策划的体液样本。我们的三个具体目标是：目的1）从12种人体体液中获得大量不同的样本（血浆、血清、脐带血、尿、脑脊液（CSF）、痰、支气管肺泡灌洗液（BAL）、唾液、精液、羊水、胚周液和卵泡液），目的2）使用现有技术的方法对体液样品中的全部exRNA进行测序，和目的3）应用适当的分析方法来鉴定人类和外源RNA，并建立代表性种族多样性美国人群的正常参考值。我们将分两个阶段进行该项目。在第一阶段（1-2年），我们将使用三种互补测序方法研究代表12种体液的约200份样本。这些研究将填补我们对人体体液中exRNA多样性的理解的主要空白，并提供有关哪些基于测序的方法对exRNA鉴定和定量最有价值的重要见解。在第二阶段（3-5年），我们将在第一阶段研究和细胞外RNA通讯计划参与者进行的其他工作的基础上，分析2,880份血浆和尿液样本，这些样本来自三项大型、种族多样性的NIH队列研究（CARDIA、REGARDS和GALA/SAGE）中招募的2,160名受试者。这些研究将有足够的把握度来建立按年龄、性别和种族分层的exRNA的正常参考范围。每一阶段都与现实的里程碑评估相关联，并可构成与联合体其他参与者协作的基础。该项目与公共卫生相关，因为它将建立健康对照参考数据，以便将exRNA作为疾病患者或有疾病风险患者的生物标志物。

DESCRIPTION (provided by applicant): Our goal is to generate reference profiles of short, long and circular non-coding regulatory exRNAs, including environmentally-derived exRNAs, in samples of 12 different body fluids from healthy humans using comprehensive RNA sequencing. We have assembled a multidisciplinary team with expertise in clinical investigation, genomics technologies, bioinformatics and statistics to achieve this goal. One major strength of this proposal is that we benefit from close interactions with other investigators participating in two UCSF-based U19 projects supported by the Common Fund Extracellular RNA Communication Program. Another major strength is that we will make use of very large sets of carefully curated body fluid samples collected from ethnically diverse populations of female and male children and adults who participated in NIH-funded cohort studies. Our three specific aims are: Aim 1) Obtain a large and diverse set of samples from 12 human body fluids (plasma, serum, cord blood, urine, cerebrospinal fluid (CSF), sputum, bronchoalveolar lavage (BAL), saliva, seminal fluid, amniotic fluid, peri-embryonic fluid and ovarian follicle fluid), Aim 2) Use state of the art approaches to sequence the full range of exRNAs in body fluid samples, and Aim 3) Apply appropriate analytical approaches to identify human and exogenous RNAs and establish normal reference values for representative ethnically-diverse US populations. We will approach the project in two phases. In phase I (years 1-2), we will study ~200 samples representing 12 body fluids using three complementary sequencing approaches. These studies will fill major gaps in our understanding of the diversity of exRNAs in human body fluids and provide important insights about which sequencing-based approaches are most valuable for exRNA identification and quantification. During phase II (years 3-5), we will build on our phase I studies and other work performed by Extracellular RNA Communication Program participants by analyzing 2,880 plasma and urine samples that have been banked from 2,160 subjects enrolled in three large, ethnically-diverse, NIH cohort studies (CARDIA, REGARDS and GALA/SAGE). These studies will be adequately powered to establish normal reference ranges for exRNAs stratified by age, sex, and ethnicity. Each phase is associated with realistic milestone assessments and can form the basis for collaboration with other consortium participants. This project is relevant to public health in thatit will establish the healthy control reference data necessary to apply exRNAs as biomarkers in patients with or at risk for disease.