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12种不同的健康人体体液样本中生成短、长和环形的非编码调控exRNAs的参考图谱，包括环境来源的exRNAs。我们已经组建了一支拥有临床研究、基因组学技术、生物信息学和统计学专业知识的多学科团队。 来实现这一目标。这项提议的一个主要优点是，我们受益于与参与共同基金细胞外RNA通讯计划支持的两个基于加州大学旧金山分校的U19项目的其他研究人员的密切互动。另一个主要优势是，我们将利用大量精心挑选的体液样本，这些样本来自参与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。我们的三个具体目标是：目的1)从12个人体体液(血浆、血清、脐带血、尿液、脑脊液、痰、支气管肺泡灌洗(BAL)、唾液、精液、羊水、胚胎周围液和卵泡液)中获取大量和多样化的样本，目的2)使用最先进的方法对体液样本中的各种exRNA进行测序，以及目的3)应用适当的分析方法来识别人类和外源RNA，并为具有代表性的不同种族的美国人群建立正常的参考值。我们将分两个阶段实施该项目。在第一阶段(1-2年)，我们将使用三种互补的测序方法研究代表12个体液的约200个样本。这些研究将填补我们对体液中exRNA多样性的了解的主要空白，并提供关于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(第3-5年)，我们将在我们的第一阶段研究和细胞外RNA通讯计划参与者所做的其他工作的基础上，通过分析2,880份血浆和尿样，这些样本来自NIH三项大型、种族多样化的队列研究(CARDIA、CORETS和GALA/SAGE)登记的2160名受试者。这些研究将有足够的力量来建立按年龄、性别和种族分层的外源RNA的正常参考范围。每个阶段都与现实的里程碑评估相关联，并可构成与其他联盟参与者合作的基础。该项目与公共卫生有关，因为它将建立必要的健康对照参考数据，以应用exRNAs作为疾病患者或有疾病风险的患者的生物标记物。

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.