Impact of circadian rhythm in obtaining reference profiles of exRNAs in healthy i

昼夜节律对获取健康人 exRNA 参考谱的影响

• 批准号: 9058134
• 负责人: IONITA Calin GHIRAN
• 金额: $ 63.69万
• 依托单位: BETH ISRAEL DEACONESS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-01 至 2019-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9058134
• 关键词: Accounting; Age; Anticoagulants; Area; Behavior; Biogenesis; Bioinformatics; Biological; Biological Markers; Blood; Blood Banks; Blood specimen; Cell membrane; Cell physiology; Cells; Circadian Rhythms; Clinical Research; Collection; Communication; Complement Activation; Complement Membrane Attack Complex; Complex; Data; Diagnosis; Diagnostic; Disease; Disease Progression; Early Diagnosis; Edetic Acid; Endocytosis; Excision; Gender; Gene Expression; Generations; Goals; Grant; Health; Heparin; Immune response; Individual; Intervention; Lipoproteins; Malignant Neoplasms; Mediating; Medicine; Membrane; Metabolic; Methods; MicroRNAs; Morphologic artifacts; Multivariate Analysis; Neoplasm Metastasis; Organ; Paracrine Communication; Patients; Physical activity; Plasma; Process; Protein Secretion; Proteins; RNA; RNA-Binding Proteins; Relapse; Reproducibility; Sampling; Signal Transduction; Sorting - Cell Movement; Source; Techniques; Time; Tissues; Transfusion; Treatment Efficacy; United States National Institutes of Health; Untranslated RNA; Vaccination; abstracting; base; cell type; circular RNA; complement system; design; differential expression; digital; disease diagnosis; extracellular; extracellular vesicles; novel; novel strategies; outcome forecast; peripheral blood; prognostic; sample collection; transcriptome sequencing; water channel; working group

DESCRIPTION (provided by applicant): Abstract A novel paradigm in paracrine signaling has recently emerged based on the findings identifying extracellular vesicles (EVs) as intercellular conveyors of biological information. Complementary to traditional modes of signaling, EV-mediated signaling appears to be critical for metabolic cooperation and coordination between cells, tissues and organs. In this context, extra-cellular RNA (exRNA), which includes microRNAs, circular RNAs, and long non-coding RNA or lncRNAs, contained both within in EVs as well as in complex with lipoproteins, have been shown by us and others to regulate gene expression and alter cell function in various cell types. Moreover, during pathological conditions such as cancer, the number and compositions of EVs and exRNA change, altering the host immune response as well as synchronizing the behavior of secondary tumors. Isolations and analysis of EVs and exRNAs both during normal and pathological conditions are critical for understanding EVs and exRNAs biogenesis and their effector functions. This information is a critical prerequisite for novel disease diagnostic and prognostic strategies, biomarker-based surveillance for disease progression, treatment efficacy and relapse. An important determinant in EVs biogenesis is represented by the complement system, which upon activation generates membrane-targeting complexes, MACs (membrane attack complex, C5b-9), which represent ungated Ca++ and water channels. Following complement activation, targeted cells either exocytose and endocytose of the MAC-containing areas of the plasma membrane generating in the process large number of EVs. Currently, the study of EVs as biological entities relevant for intercellular signaling and disease diagnosis is based on the assumption that the biogenesis of EVs and exRNA happen at a steady state rate, being modified mostly by the healthy/diseased status of the host. Our preliminary data strongly suggest that that may not be the case. Our results show that the tissue-origin, number, protein and RNA composition, of EVs isolated by standard techniques depend not just on the blood collection methods, but also on the time of day the blood samples were collected. In this application, we propose to establish a baseline for the quality, quantity and composition of extracellular RNA, taking into account the potential artifacts induced by the: 1) blood collection methods, 2) time of collection, 3) physical activity. Our results obtained with fresh samples, will be cross-validated using stored plasma samples from the Center for Clinical Research and Division of Transfusion Medicine (NIH Blood Bank).

描述（由申请人提供）： 摘要：基于将细胞外囊泡（EV）确定为细胞间生物信息传递者的发现，最近出现了旁分泌信号传导的新范例。作为传统信号传导模式的补充，EV介导的信号传导似乎对于细胞、组织和器官之间的代谢合作和协调至关重要。在这种情况下，我们和其他人已证明细胞外 RNA (exRNA)（包括 microRNA、环状 RNA 和长链非编码 RNA 或 lncRNA）可以调节 EV 中以及与脂蛋白的复合物。基因表达并改变各种细胞类型的细胞功能。此外，在癌症等病理条件下，EV 和 exRNA 的数量和组成会发生变化，从而改变宿主免疫反应并同步继发性肿瘤的行为。正常和病理条件下 EV 和 exRNA 的分离和分析对于了解 EV 和 exRNA 的生物发生及其效应子功能至关重要。这些信息是新的疾病诊断和预后策略、基于生物标志物的疾病进展、治疗效果和复发监测的关键先决条件。 EV 生物合成中的一个重要决定因素是补体系统，该系统激活后会生成膜靶向复合物 MAC（膜攻击复合物，C5b-9），代表非门控 Ca++ 和水通道。补体激活后，靶细胞对质膜的 MAC 区域进行胞吐或内吞，在此过程中产生大量 EV。目前，对 EV 作为与细胞间信号传导和疾病诊断相关的生物实体的研究基于这样的假设：EV 和 exRNA 的生物发生以稳态速率发生，主要受到宿主健康/疾病状态的影响。我们的初步数据强烈表明情况可能并非如此。我们的结果表明，通过标准技术分离的 EV 的组织来源、数量、蛋白质和 RNA 组成不仅取决于血液采集方法，还取决于血液样本采集的时间。在此应用中，我们建议建立细胞外 RNA 的质量、数量和组成的基线，同时考虑以下因素引起的潜在伪影：1) 血液采集方法，2) 采集时间，3) 体力活动。我们用新鲜样品获得的结果将 使用临床研究中心和输血医学部（NIH 血库）储存的血浆样本进行交叉验证。