Effect of methodological and biological variability on molecular profiling of extracellular vesicles in cancer detection

方法学和生物学变异对癌症检测中细胞外囊泡分子谱的影响

• 批准号: 10115636
• 负责人: IONITA Calin GHIRAN
• 金额: $ 68万
• 依托单位: NORTHEASTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-03-15 至 2023-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10115636
• 关键词: Age; Anticoagulants; Behavior; Biogenesis; Biological; Biological Assay; Biological Markers; Biology; Blood; Blood specimen; Cancer Detection; Cancer Diagnostics; Cancer Patient; Cell physiology; Cells; Clinical Medicine; Collection; Colorectal Cancer; Data; Detection; Diagnostic; Disease; Disease Progression; Disease Surveillance; Diurnal Rhythm; Equipment; Excision; Exercise; Fibrinogen; Gender; Gene Expression; Glycopeptides; Glycoproteins; Goals; Grant; Health; Immune response; Individual; Lipids; Location; Malignant Neoplasms; Malignant neoplasm of prostate; Measurement; Methodology; Methods; Molecular; Molecular Profiling; Neoplasm Metastasis; Organ; Paracrine Communication; Pathologic; Patients; Peptides; Physiological; Plasma; Post-Translational Protein Processing; Preparation; Protein Glycosylation; Proteins; Proteomics; Publishing; RNA; Relapse; Reproducibility; Sampling; Screening for cancer; Serum; Signal Transduction; Source; Specificity; Stable Isotope Labeling; Statistical Data Interpretation; Statistical Methods; Surface; Techniques; Time; Tissues; Treatment Efficacy; Vaccination; Validation; Variant; analytical method; base; biomarker discovery; cell type; circadian; disease diagnosis; extracellular vesicles; glycoproteomics; glycosylation; individual variation; insight; nanoflow cytometry; novel; pancreatic cancer patients; personalized medicine; prognostic; treatment response

Project Summary A novel paradigm in paracrine signaling has recently emerged based on the findings identifying extracellular vesicles (EVs) as intercellular conveyors of biological information both in normal and pathological conditions such as cancer. EVs and their cargo 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 alter the host immune response as well as synchronize the behavior of secondary tumors. Isolation and molecular profiling of EVs (i.e. RNAs, proteins, post-translational modifications, lipids, metabolites) both in health and disease are critical for understanding EVs' biogenesis and effector functions. Currently, the study of EVs as biological entities relevant for intracellular signaling and disease diagnosis is based on the assumption that the biogenesis and removal of EVs happen at a steady state rate, being modified mostly by the healthy/diseased status of the host. Our published results show that that is not the case. Our data indicate that the tissue-origin, number, size distribution, as well as protein, lipid, metabolite and RNA composition of EVs isolated by standard techniques depend not just on the blood collection methods, but also on the time of the day the blood samples were collected. Therefore, systematic assessment of these factors and other sources of variability in EV profiles are important for enabling basic biology, clinical and personalized medicine applications. Although it is beyond the purpose of this grant, the long-term goal of our team is to establish reproducible methods for blood collection and sample processing that would allow us to identify the specific molecular EV signatures during the day/night cycle, with the aim of pinpointing the ideal, organ-specific times for blood collection, which would increase the reliability and specificity of early cancer detection. We believe that establishing biological and methodological baselines are absolutely vital for correctly comparing proteomics and glycomics data obtained in various studies, as well as interpreting the data obtained from patients' samples.

项目概要 基于识别细胞外信号的发现，最近出现了一种旁分泌信号传导的新范例 囊泡（EV）作为正常和病理条件下生物信息的细胞间传递者 例如癌症。我们和其他人已经证明电动汽车及其货物可以调节基因表达并改变 各种细胞类型的细胞功能。此外，在癌症等病理状况下，数量和 EV 的成分改变宿主免疫反应并同步继发性肿瘤的行为。 EV 的分离和分子分析（即 RNA、蛋白质、翻译后修饰、脂质、 代谢物）在健康和疾病中对于理解 EV 的生物发生和效应器功能至关重要。 目前，EVs作为与细胞内信号传导和疾病诊断相关的生物实体的研究正在进行中。 基于 EV 的生物发生和去除以稳态速率发生的假设， 主要由宿主的健康/患病状态改变。我们发表的结果表明，这不是 案件。我们的数据表明，组织来源、数量、大小分布，以及蛋白质、脂质、代谢物和 通过标准技术分离的 EV 的 RNA 组成不仅取决于血液采集方法，还取决于 也在一天中的某个时间采集血液样本。因此，系统地评估这些 EV 特征中的因素和其他变异来源对于实现基础生物学、临床和 个性化医疗应用。 虽然这超出了本次资助的目的，但我们团队的长期目标是建立可重复的 血液采集和样品处理方法使我们能够识别特定的分子 EV 昼/夜周期中的特征，目的是确定理想的、特定于器官的血液时间 收集，这将提高早期癌症检测的可靠性和特异性。我们相信 建立生物学和方法学基线对于正确比较蛋白质组学绝对至关重要 和在各种研究中获得的糖组学数据，以及解释从患者获得的数据 样品。